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Volume 22,Issue 4,2015 Table of Contents
2015, 22(4):413-419.
DOI: 10.3872/j.issn.1007-385X.2015.04.001
Abstract:
In recent years, the breakthroughs of targeting agents for tumor therapies and personalized molecular diagnosis have prompted cancer treatment into the era of precision medicine. However, established assessment for solid tumors treatment such as the WHO or RECIST evaluation criteria could not reflex precisely the efficacy of the targeted therapeutics and the survival benefits they provided to patients. Therefore, it is extremely urgent to explore and establish a new response assessment criteria for targeted tumor therapeutics. This article reviewed the history of the efficacy assessment criteria for anti-tumor drugs and current opinions on the subject. I also introduced the evolution and development of evaluation criteria for various targeted therapeutics toward different tumors, and focused on the new criteria for evaluating immuno-related therapeutics. Finally, I discussed in detail the proposed evaluation criteria, its definition, guiding principle and clinical application. It is conceivable that the continuous improvements in evaluating targeted therapeutics for solid tumors will bring transformation to personalized cancer treatment.
In recent years, the breakthroughs of targeting agents for tumor therapies and personalized molecular diagnosis have prompted cancer treatment into the era of precision medicine. However, established assessment for solid tumors treatment such as the WHO or RECIST evaluation criteria could not reflex precisely the efficacy of the targeted therapeutics and the survival benefits they provided to patients. Therefore, it is extremely urgent to explore and establish a new response assessment criteria for targeted tumor therapeutics. This article reviewed the history of the efficacy assessment criteria for anti-tumor drugs and current opinions on the subject. I also introduced the evolution and development of evaluation criteria for various targeted therapeutics toward different tumors, and focused on the new criteria for evaluating immuno-related therapeutics. Finally, I discussed in detail the proposed evaluation criteria, its definition, guiding principle and clinical application. It is conceivable that the continuous improvements in evaluating targeted therapeutics for solid tumors will bring transformation to personalized cancer treatment.
Li Zhenhai
,
Wu Hongping
,
Xu Zenghui
,
Lü Saiqun
,
Shi Junxia
,
Liu Pinyi
,
Li Linfang
,
Jin Huajun
,
Wu Mengchao
,
Qian Qijun
2015, 22(4):420-426.
DOI: 10.3872/j.issn.1007-385X.2015.04.002
Abstract:
Objective:To construct anadenovirus mediated dual-luciferase reportersystemthat can be used for screening promoters with high activity in a broad of cells. Methods: The testing promoter controlled expressioncassette of Firefly luciferase and CMV promoter controlled expressioncassette of Renilla luciferase were cloned into the E1 and E3 region of adenoviral vector respectively. The capacity of the system was assessed in the following three aspects: the activities of each promoter detected at different MOI, the variance within the same group, and the availability of the system for examining the promoter activity in cell lines that were hard to be transfected. Finally, the activities of CAG, CASI and a new CCAU promoters were examined by the system in 9 cell lines to find the promoter with high and broad-spectrum activity.Results: The adenovirus mediated dual-luciferase reporters system was successfully constructed. The activities of Rluc and Fluc of individual promoter had no significant difference at different MOI conditions(P>0.05), and the variations in each test group were small. The system worked effectively in K562, Jurkat, and primary skin cells, which are hard to be transfected. We found that the CCAU promoter produced higher luciferase activity than the CAG and CASI promoters in 9 cell lines tested (P<0.05 and P<0.01 respectively). Conclusion: The adenovirus mediated dual-luciferase reporters is an easy-to-use and reliable assay system suitable for assessing promoter activity. The new designed promoter CCAU has broad-spectrum strongactivity and can be used in transgenic over-expression.
Objective:To construct anadenovirus mediated dual-luciferase reportersystemthat can be used for screening promoters with high activity in a broad of cells. Methods: The testing promoter controlled expressioncassette of Firefly luciferase and CMV promoter controlled expressioncassette of Renilla luciferase were cloned into the E1 and E3 region of adenoviral vector respectively. The capacity of the system was assessed in the following three aspects: the activities of each promoter detected at different MOI, the variance within the same group, and the availability of the system for examining the promoter activity in cell lines that were hard to be transfected. Finally, the activities of CAG, CASI and a new CCAU promoters were examined by the system in 9 cell lines to find the promoter with high and broad-spectrum activity.Results: The adenovirus mediated dual-luciferase reporters system was successfully constructed. The activities of Rluc and Fluc of individual promoter had no significant difference at different MOI conditions(P>0.05), and the variations in each test group were small. The system worked effectively in K562, Jurkat, and primary skin cells, which are hard to be transfected. We found that the CCAU promoter produced higher luciferase activity than the CAG and CASI promoters in 9 cell lines tested (P<0.05 and P<0.01 respectively). Conclusion: The adenovirus mediated dual-luciferase reporters is an easy-to-use and reliable assay system suitable for assessing promoter activity. The new designed promoter CCAU has broad-spectrum strongactivity and can be used in transgenic over-expression.
2015, 22(4):427-431.
DOI: 10.3872/j.issn.1007-385X.2015.04.003
Abstract:
Objective:To investigate whether c-Met inhibitor SU11274 can reverse resistance to Erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), induced by HGF in non-small cell lung cancer cells with different EGFR gene types. Methods: NSCLC cells with different EGFR gene types, including PC9 (EGFR-activating mutant), H292 (EGFR-wild type), and A549 (EGFR-wild type) were utilized in the study. The experiments for each cell line consisted of six different treatment groups: C group (control), H group (HGF), E group (Erlotinib), S group (SU11274), EH group (Erlotinib+HGF), and ESH group (Erlotinib+SU11274+HGF). Their effects on cell survival and apoptosis were measured by MTT assay and flow cytometry (FCM). The activation of c-Met, Stat3, Akt, and Erk1/2 protein were examined by immunoblotting. Results: Erlotinib inhibited growth of the three cells lines in a dose-dependent manner, and the inhibition was effectively blocked by HGF. The presence of SU11274 significantly decreased the survival rates of cells exposed to HGF and Erlotinib (P<0.05). Apoptosis in cells treated with Erlotinib, SU11274, and HGF was also markedly increased compared with these treated with Erlotinib and HGF only (P<0.05). Similarly, the levels of p-Met, p-Stat3, p-Akt, and p-Erk1/2 in the HES group were significantly lower than that in the HE group (P<0.05). Conclusion: SU11274 reversed HGF-induced resistance to Erlotinib in non-small lung cancer cells with different EGFR gene type, likely due to the inhibition of HGF-induced activation of c-Met and its down streams signaling pathways.
Objective:To investigate whether c-Met inhibitor SU11274 can reverse resistance to Erlotinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), induced by HGF in non-small cell lung cancer cells with different EGFR gene types. Methods: NSCLC cells with different EGFR gene types, including PC9 (EGFR-activating mutant), H292 (EGFR-wild type), and A549 (EGFR-wild type) were utilized in the study. The experiments for each cell line consisted of six different treatment groups: C group (control), H group (HGF), E group (Erlotinib), S group (SU11274), EH group (Erlotinib+HGF), and ESH group (Erlotinib+SU11274+HGF). Their effects on cell survival and apoptosis were measured by MTT assay and flow cytometry (FCM). The activation of c-Met, Stat3, Akt, and Erk1/2 protein were examined by immunoblotting. Results: Erlotinib inhibited growth of the three cells lines in a dose-dependent manner, and the inhibition was effectively blocked by HGF. The presence of SU11274 significantly decreased the survival rates of cells exposed to HGF and Erlotinib (P<0.05). Apoptosis in cells treated with Erlotinib, SU11274, and HGF was also markedly increased compared with these treated with Erlotinib and HGF only (P<0.05). Similarly, the levels of p-Met, p-Stat3, p-Akt, and p-Erk1/2 in the HES group were significantly lower than that in the HE group (P<0.05). Conclusion: SU11274 reversed HGF-induced resistance to Erlotinib in non-small lung cancer cells with different EGFR gene type, likely due to the inhibition of HGF-induced activation of c-Met and its down streams signaling pathways.
2015, 22(4):432-437.
DOI: 10.3872/j.issn.1007-385X.2015.04.004
Abstract:
Objective:To explore the role ofSTAT3 signaling pathway in the interaction of glioma cells and human brain microvascular endothelial cells (HBMEC). Methods: The secretion of IL-6 from HBMEC co-cultured with VEGF，A172, or U251 cells by Elisa. Immunoblotting was performed to examine STAT3 and its phosphorylation status inglioma cells after exposed to IL-6(50 ng/ml), co-cultured with the endothelial cells, orHBMEC+AZD1480. CCK-8 and Transwell assays were used to measure the effect of IL-6 (50 ng/ml), HBMEC, and HBMEC+AZD1480 on the proliferation and invasion of glioma cells A172 and U251. Results: Glioma cells and VEGF promoted the secretion of the IL-6 bythe endothelial cells(P<0.01). The level of pSTAT3 in A172 and U251 was significantly increased after exposed to IL-6， HBMEC， or HBMEC+AZD1480 (P<0.01). Moreover, AZD1480 reduced STAT3 phosphorylationin A172 and U251 co-cultured with HBME(P<0.01). IL-6 (50ng/ml) and co-culturing with the endothelial cells enhanced the growth and invasion of the glioma cells(P<0.01), where as AZD1480 suppressed the growth and invasion of the A172 and U251 cells (P<001). Conclusion: STAT3 activation is involved in the interaction between glioma cells and the human brain microvascular endothelial cells.
Objective:To explore the role ofSTAT3 signaling pathway in the interaction of glioma cells and human brain microvascular endothelial cells (HBMEC). Methods: The secretion of IL-6 from HBMEC co-cultured with VEGF，A172, or U251 cells by Elisa. Immunoblotting was performed to examine STAT3 and its phosphorylation status inglioma cells after exposed to IL-6(50 ng/ml), co-cultured with the endothelial cells, orHBMEC+AZD1480. CCK-8 and Transwell assays were used to measure the effect of IL-6 (50 ng/ml), HBMEC, and HBMEC+AZD1480 on the proliferation and invasion of glioma cells A172 and U251. Results: Glioma cells and VEGF promoted the secretion of the IL-6 bythe endothelial cells(P<0.01). The level of pSTAT3 in A172 and U251 was significantly increased after exposed to IL-6， HBMEC， or HBMEC+AZD1480 (P<0.01). Moreover, AZD1480 reduced STAT3 phosphorylationin A172 and U251 co-cultured with HBME(P<0.01). IL-6 (50ng/ml) and co-culturing with the endothelial cells enhanced the growth and invasion of the glioma cells(P<0.01), where as AZD1480 suppressed the growth and invasion of the A172 and U251 cells (P<001). Conclusion: STAT3 activation is involved in the interaction between glioma cells and the human brain microvascular endothelial cells.
2015, 22(4):438-442.
DOI: 10.3872/j.issn.1007-385X.2015.04.005
Abstract:
Objective:To investigate the effect of 4EGI-1 on AKT activity in melanoma cell A375 and its role in 4EGI-1-induced growth inhibition of the cells. Methods: After treated with indicated concentrations of 4EGI-1（20, 40, 60 μmol/L）for 12 h, A375 cells were lysed to analyze AKT phosphorylation by using immunoblotting. A375 cells were also treated with 4EGI-1 and BEZ235 alone or in combination to assess their effects on cell proliferation through the CCK-8 assay. Their influences on cell cycle were determined using flow cytometry and the expression of Cyclin D1 was examined by immunoblotting.Results: 4EGI-1 induced AKT phosphorylation in A375 cells dose-dependently. While the inhibition rates of 4EGI-1 and BEZ235 on A375 cells were (7.6±1.2)% and (2.4±3.1)% respectively, the rate increased significantly to (19.8±4.3)% when both compounds were used together. The percentage of cells in S phase cells were 2565% and 25.82% after treated with 4EGI-1 or BEZ235. It decreased to 13.08% in A375 cells exposed to both 4EGI-1 and BEZ235, indicating that BEZ235 enhanced the inhibitory effect of 4EGI-1 on A375 growth. BEZ235 also enhanced the inhibitory effect of 4EGI-1 on cyclin D1 expression. The relative expression levels of cyclin D1 in 4EGI-1 group and BEZ235 group were 0.81±0.04 and 0.04±0.76 respectively, it was reduced to 0.25±0.03 when the cells were cultured in the presence of both compounds. Conclusion: Akt phosphorylation induced by 4EGI-1 counteracts its growth inhibition on A375 cell. Combination of 4EGI-1 and Akt inhibitor is a more effective strategy to inhibit the growth of A375 cells.
Objective:To investigate the effect of 4EGI-1 on AKT activity in melanoma cell A375 and its role in 4EGI-1-induced growth inhibition of the cells. Methods: After treated with indicated concentrations of 4EGI-1（20, 40, 60 μmol/L）for 12 h, A375 cells were lysed to analyze AKT phosphorylation by using immunoblotting. A375 cells were also treated with 4EGI-1 and BEZ235 alone or in combination to assess their effects on cell proliferation through the CCK-8 assay. Their influences on cell cycle were determined using flow cytometry and the expression of Cyclin D1 was examined by immunoblotting.Results: 4EGI-1 induced AKT phosphorylation in A375 cells dose-dependently. While the inhibition rates of 4EGI-1 and BEZ235 on A375 cells were (7.6±1.2)% and (2.4±3.1)% respectively, the rate increased significantly to (19.8±4.3)% when both compounds were used together. The percentage of cells in S phase cells were 2565% and 25.82% after treated with 4EGI-1 or BEZ235. It decreased to 13.08% in A375 cells exposed to both 4EGI-1 and BEZ235, indicating that BEZ235 enhanced the inhibitory effect of 4EGI-1 on A375 growth. BEZ235 also enhanced the inhibitory effect of 4EGI-1 on cyclin D1 expression. The relative expression levels of cyclin D1 in 4EGI-1 group and BEZ235 group were 0.81±0.04 and 0.04±0.76 respectively, it was reduced to 0.25±0.03 when the cells were cultured in the presence of both compounds. Conclusion: Akt phosphorylation induced by 4EGI-1 counteracts its growth inhibition on A375 cell. Combination of 4EGI-1 and Akt inhibitor is a more effective strategy to inhibit the growth of A375 cells.
2015, 22(4):443-447.
DOI: 10.3872/j.issn.1007-385X.2015.04.006
Abstract:
Objective:To study the effect of cisplatin and paclitaxel on the cytotoxic activity of cytokine-induced killer (CIK) cells toward human lung adenocarcinoma cell A549 and further explore the possible molecular mechanism involved. Methods: The IC50 of cisplatin and paclitaxel in inhibiting A549 cells were measured by using MTT assay. Cytotoxicity of CIK cells toward A549 cells that were pre-cultured with cisplatin or paclitaxel was measured through LDH releasing assay. The expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on A549 cells were analyzed by flow cytometry. The level of DNA damage response genes (ATM, ATR, CHK1, CHK2, P53) were assessed by fluorescent quantitative PCR.Results: The IC50 of cisplatin and paclitaxel in inhibiting A594 were 70 μg/ml and 39 μg/ml respectively. Cytotoxicity of CIK cells against A549 cells was significantly increased after the cells were pre-treatment with cisplatin (70 μg/ml) or paclitaxel (39 μg/ml) (P<0.05). Expression of MICA, MICB, ULBP2, and ULBP3 on A549 cells was significantly increased by the cisplatin or paclitaxel pretreatment (P<0.05), where as the level of ULBP1 was decreased after the expose (P<0.05). Furthermore, ATM expression in A594 cells was decreasedby cisplatin treatment \[(3.23±1.62)×10-6 vs (5.49±3.91)×10-8，P<0.05\], whereas the level of P53 mRNA elevated significantly after treatmentwith paclitaxel \[(14.90±5.49)×10-6vs (3.68±2.82)×10-6，P<0.05\]. Conclusion: The results indicated that cisplatinand or paclitaxel can enhance the cytotoxic activity of CIK cells toward lung cancer cells. It is conceivable that they activate DNA-damage response genes, which in turnupregulate the expression of NKG2D ligands that make cells more susceptible to CIK cells.
Objective:To study the effect of cisplatin and paclitaxel on the cytotoxic activity of cytokine-induced killer (CIK) cells toward human lung adenocarcinoma cell A549 and further explore the possible molecular mechanism involved. Methods: The IC50 of cisplatin and paclitaxel in inhibiting A549 cells were measured by using MTT assay. Cytotoxicity of CIK cells toward A549 cells that were pre-cultured with cisplatin or paclitaxel was measured through LDH releasing assay. The expression of NKG2D ligands (MICA, MICB, ULBP1, ULBP2, ULBP3) on A549 cells were analyzed by flow cytometry. The level of DNA damage response genes (ATM, ATR, CHK1, CHK2, P53) were assessed by fluorescent quantitative PCR.Results: The IC50 of cisplatin and paclitaxel in inhibiting A594 were 70 μg/ml and 39 μg/ml respectively. Cytotoxicity of CIK cells against A549 cells was significantly increased after the cells were pre-treatment with cisplatin (70 μg/ml) or paclitaxel (39 μg/ml) (P<0.05). Expression of MICA, MICB, ULBP2, and ULBP3 on A549 cells was significantly increased by the cisplatin or paclitaxel pretreatment (P<0.05), where as the level of ULBP1 was decreased after the expose (P<0.05). Furthermore, ATM expression in A594 cells was decreasedby cisplatin treatment \[(3.23±1.62)×10-6 vs (5.49±3.91)×10-8，P<0.05\], whereas the level of P53 mRNA elevated significantly after treatmentwith paclitaxel \[(14.90±5.49)×10-6vs (3.68±2.82)×10-6，P<0.05\]. Conclusion: The results indicated that cisplatinand or paclitaxel can enhance the cytotoxic activity of CIK cells toward lung cancer cells. It is conceivable that they activate DNA-damage response genes, which in turnupregulate the expression of NKG2D ligands that make cells more susceptible to CIK cells.
2015, 22(4):448-453.
DOI: 10.3872/j.issn.1007-385X.2015.04.007
Abstract:
Objective:To investigate the effect and mechanism of Bestrophin 3 on the apoptosis of human hepatocellular carcinoma cell HepG2. Methods: HepG2 cells were transfected with various multiplicity of infection (MOI=10, 20, 40 and 80) of adenovirus expressing Bestrophin 3, LacZ, or control. The infection efficiency of the adenovirus was measured by Western blotting. Proliferation and apoptosis rates of the transfected cells were measured with CCK-8 assay and flow cytometry. Effects of Bestrophin 3 on the expression of Bcl-2 and Bax and the release of cytochrome C (Cyt C) were determined by using immunoblotting. Influence of Bestrophin 3 overexpression on mitochondrial membrane potential was assessed by JC-1 and fluorescence microscopy. Results: The expression of Bestrophin 3 reached the maximal level after adenoviral infection at 40 MOI. Compared with the control and LacZ-expressing groups, the cell viability of the Bestrophin 3-expressing group was significantly reduced \[(79.37±1.76)% vs (98.67±3.02)% and (99.67±3.25)%，P<005\], whereas the apoptosis rate was significantly increased \[(29.47±2.37)% vs （5.47±0.37）% and (4.95±044）%，P<0.05\]. In the cells infected with the adenovirus expressing Bestrophin 3, there were decreased level of Bcl-2 and increased level of Bax, leading to a significant reduction of Bcl-2/Bax ratio. In HepG2 cells overexpressing Bestrophin 3, the mitochondrial membrane potential was also significantly lower compared with that of controls (1.00±0.00)% vs 0.64±0.09%, P<0.05), and Cyt C content was reduced in mitochondria and elevated in cytoplasm (all P<0.05). Conclusion: Enforced expression of Bestrophin 3 increases the apoptosis of HepG2 through reducing Bcl-2/Bax ratio and promoting cytochrome C release from the mitochondria.
Objective:To investigate the effect and mechanism of Bestrophin 3 on the apoptosis of human hepatocellular carcinoma cell HepG2. Methods: HepG2 cells were transfected with various multiplicity of infection (MOI=10, 20, 40 and 80) of adenovirus expressing Bestrophin 3, LacZ, or control. The infection efficiency of the adenovirus was measured by Western blotting. Proliferation and apoptosis rates of the transfected cells were measured with CCK-8 assay and flow cytometry. Effects of Bestrophin 3 on the expression of Bcl-2 and Bax and the release of cytochrome C (Cyt C) were determined by using immunoblotting. Influence of Bestrophin 3 overexpression on mitochondrial membrane potential was assessed by JC-1 and fluorescence microscopy. Results: The expression of Bestrophin 3 reached the maximal level after adenoviral infection at 40 MOI. Compared with the control and LacZ-expressing groups, the cell viability of the Bestrophin 3-expressing group was significantly reduced \[(79.37±1.76)% vs (98.67±3.02)% and (99.67±3.25)%，P<005\], whereas the apoptosis rate was significantly increased \[(29.47±2.37)% vs （5.47±0.37）% and (4.95±044）%，P<0.05\]. In the cells infected with the adenovirus expressing Bestrophin 3, there were decreased level of Bcl-2 and increased level of Bax, leading to a significant reduction of Bcl-2/Bax ratio. In HepG2 cells overexpressing Bestrophin 3, the mitochondrial membrane potential was also significantly lower compared with that of controls (1.00±0.00)% vs 0.64±0.09%, P<0.05), and Cyt C content was reduced in mitochondria and elevated in cytoplasm (all P<0.05). Conclusion: Enforced expression of Bestrophin 3 increases the apoptosis of HepG2 through reducing Bcl-2/Bax ratio and promoting cytochrome C release from the mitochondria.
2015, 22(4):454-458.
DOI: 10.3872/j.issn.1007-385X.2015.04.008
Abstract:
Objective:To investigate the effects of siRNA-mediated Semaphorin 4D (Sema 4D) gene silence on the proliferation and migration of breast cancer MDA-MB-231 cells. Methods:Small inference RNA targeting different regions of Sema 4D gene were transfected into MDA-MB-231 cells by lipofectamine 2000. The most effective siRNA in knockdown Sema 4D was screened by using quantitative PCR and Western blotting. The changes of cell proliferation and migration ability after Sema 4D knockdown were examined by CCK-8 and transwell assays. Results: MDA-MB-231 cells transfected with Sema 4D siRNA had significantly decreased expression of Sema 4D, especially in these transfected with siRNA-C (P<005). Compared with the blank and scrambled siRNA transfection groups, siRNA-C transfection significantly inhibited the proliferation of MDA-MB-231 cells (P<0.05) and their migration ability （numbers of migration cell：\[19620±904\] and \[186.40±6.69\] vs \[105.60±12.07\]，P<0.05). Conclusion: Down-regulation of Sema 4D by siRNA significantly inhibits the proliferation and migration of MDA-MB-231 cells, suggesting Sema 4Dis a novel target for breast cancer therapy.
Objective:To investigate the effects of siRNA-mediated Semaphorin 4D (Sema 4D) gene silence on the proliferation and migration of breast cancer MDA-MB-231 cells. Methods:Small inference RNA targeting different regions of Sema 4D gene were transfected into MDA-MB-231 cells by lipofectamine 2000. The most effective siRNA in knockdown Sema 4D was screened by using quantitative PCR and Western blotting. The changes of cell proliferation and migration ability after Sema 4D knockdown were examined by CCK-8 and transwell assays. Results: MDA-MB-231 cells transfected with Sema 4D siRNA had significantly decreased expression of Sema 4D, especially in these transfected with siRNA-C (P<005). Compared with the blank and scrambled siRNA transfection groups, siRNA-C transfection significantly inhibited the proliferation of MDA-MB-231 cells (P<0.05) and their migration ability （numbers of migration cell：\[19620±904\] and \[186.40±6.69\] vs \[105.60±12.07\]，P<0.05). Conclusion: Down-regulation of Sema 4D by siRNA significantly inhibits the proliferation and migration of MDA-MB-231 cells, suggesting Sema 4Dis a novel target for breast cancer therapy.
Zheng Jie
,
Jiang Longwei
,
Yao Lu
,
Ai Yueqin
,
Zhang Yan
,
Huang Weiqian
,
Gao Yanrong
,
Zhang Chuang
,
Zhao Hua
,
Hu Jianhua
,
Jia Shaochang
,
Yu Shuyong
2015, 22(4):459-464.
DOI: 10.3872/j.issn.1007-385X.2015.04.009
Abstract:
Objective:To evaluate the safety and efficacy of combination therapy with dendritic cells (DCs) and cytokine-induced killer (CIK) cells in the treatment of advanced colorectal cancer (CRC). Methods: Peripheral blood mononuclear cells (PBMCs) were collected from 142 patients with stage Ⅲ~Ⅳ CRC who were admitted to the Tumor Biotherapy Center of the No. 81 Hospital of PLA in Nanjing from August, 2011 to December, 2014. The cells were cultured ex vivo to generate DC and CIK cells. After sensitized with cell lysates from colon cancer cells Colo-320 or rectal cancer cells HCT-116, the DCs were transfused to CRC patients after combined with the CIK cells. T-lymphocyte subsets, serum CEA level, and clinical features were evaluated before and after the combined DC and CIK treatment. Results: Following the combined DC and CIK immunotherapy in patients with advanced CRC for 40 months, the overall response rate was 162%, the disease control rate was 60.0%, one-year overall survival rate was 47%, and both two-year and three-year overall survival rate was maintained at 31%. No significant alterations in T-lymphocyte subsets, CD4+/CD8+ ratio, and CEA level were found in these patients after the combination therapy. Single-factor analysis indicated that tumor stage (P=0.015), the frequency of immunotherapy (P=0.037), and CEA value (P=0.037) affected significantly the survival period of these CRC patients. Multivariate Cox model indicated that frequency of the combined DC and CIK immunotherapy (P=0.024) and age (P=0.015) associated significantly with the risk of cancer-related death in these CRC patients. Conclusion: The combined autologous DC/CIK immunotherapy is a safe and feasible therapeutic approach and it may improve the long-term survival rate in patients with stage Ⅲ~Ⅳ CRC.
Objective:To evaluate the safety and efficacy of combination therapy with dendritic cells (DCs) and cytokine-induced killer (CIK) cells in the treatment of advanced colorectal cancer (CRC). Methods: Peripheral blood mononuclear cells (PBMCs) were collected from 142 patients with stage Ⅲ~Ⅳ CRC who were admitted to the Tumor Biotherapy Center of the No. 81 Hospital of PLA in Nanjing from August, 2011 to December, 2014. The cells were cultured ex vivo to generate DC and CIK cells. After sensitized with cell lysates from colon cancer cells Colo-320 or rectal cancer cells HCT-116, the DCs were transfused to CRC patients after combined with the CIK cells. T-lymphocyte subsets, serum CEA level, and clinical features were evaluated before and after the combined DC and CIK treatment. Results: Following the combined DC and CIK immunotherapy in patients with advanced CRC for 40 months, the overall response rate was 162%, the disease control rate was 60.0%, one-year overall survival rate was 47%, and both two-year and three-year overall survival rate was maintained at 31%. No significant alterations in T-lymphocyte subsets, CD4+/CD8+ ratio, and CEA level were found in these patients after the combination therapy. Single-factor analysis indicated that tumor stage (P=0.015), the frequency of immunotherapy (P=0.037), and CEA value (P=0.037) affected significantly the survival period of these CRC patients. Multivariate Cox model indicated that frequency of the combined DC and CIK immunotherapy (P=0.024) and age (P=0.015) associated significantly with the risk of cancer-related death in these CRC patients. Conclusion: The combined autologous DC/CIK immunotherapy is a safe and feasible therapeutic approach and it may improve the long-term survival rate in patients with stage Ⅲ~Ⅳ CRC.
2015, 22(4):465-471.
DOI: 10.3872/j.issn.1007-385X.2015.04.010
Abstract:
Objective:To investigate the clinicopathologic features of colorectal cancers with CpG island methylation phenotype (CIMP) by using droplet digital PCR, and to evaluate the feasibility of determining the CIMP status using plasma free DNA. Methods: Two hundred sixteen tumor and paired plasma samples were collected from patients with colorectal cancer enrolled into Changhai Hospital from 2008 to 2012. DNA was extracted from tumor tissues and plasmas and underwent bisulfate conversion. CIMP was determined by using digital PCR for the five genes CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. P53 mutation was detected by immunohistochemical analysis. K-RAS and BRAF mutations were assessed by nest PCR. Results: Among the 216 colorectal cancers, 17 (7.9%) had BRAF mutations, 96 (44.4%) contained K-RAS mutations, 112 (51.9%) were P53- IHC (+), and 68 (31.5%) were CIMP positive (≧3 gene loci were hypermethylated). Most of the cancers with BRAF mutation (82.3%, 14/17) were CIMP(+), whereas only 94% (9/96) of the cancer with K-RAS mutations and 7.1% (8/112) of cancers harboring P53 mutations (+) were CIMP(+). Compared to CIMP(-) tumors, the unique clinical pathological features of the CIMP(+) tumors included: (1) more likely to happen in the proximal locations; (2) more likely to be mucinous carcinoma; (3) higher degree of differentiation. The overall survival period of colorectal patients with CIMP(+) was significantly shorter than that with CIMP(-). When plasma free DNA from these patients was used for CIMP analysis, the consistency, sensitivity and specificity were 93.4%, 87.2% and 100% respectively compared with the assay using tumor DNA.Conclusion: Patients with CIMP(+) colorectal cancer have unique clinical pathological characteristics and poor prognosis. When tumor tissues are not available, CIMP analysis with plasma free DNA using digital PCR is a feasible alternative.
Objective:To investigate the clinicopathologic features of colorectal cancers with CpG island methylation phenotype (CIMP) by using droplet digital PCR, and to evaluate the feasibility of determining the CIMP status using plasma free DNA. Methods: Two hundred sixteen tumor and paired plasma samples were collected from patients with colorectal cancer enrolled into Changhai Hospital from 2008 to 2012. DNA was extracted from tumor tissues and plasmas and underwent bisulfate conversion. CIMP was determined by using digital PCR for the five genes CACNA1G, IGF2, NEUROG1, RUNX3 and SOCS1. P53 mutation was detected by immunohistochemical analysis. K-RAS and BRAF mutations were assessed by nest PCR. Results: Among the 216 colorectal cancers, 17 (7.9%) had BRAF mutations, 96 (44.4%) contained K-RAS mutations, 112 (51.9%) were P53- IHC (+), and 68 (31.5%) were CIMP positive (≧3 gene loci were hypermethylated). Most of the cancers with BRAF mutation (82.3%, 14/17) were CIMP(+), whereas only 94% (9/96) of the cancer with K-RAS mutations and 7.1% (8/112) of cancers harboring P53 mutations (+) were CIMP(+). Compared to CIMP(-) tumors, the unique clinical pathological features of the CIMP(+) tumors included: (1) more likely to happen in the proximal locations; (2) more likely to be mucinous carcinoma; (3) higher degree of differentiation. The overall survival period of colorectal patients with CIMP(+) was significantly shorter than that with CIMP(-). When plasma free DNA from these patients was used for CIMP analysis, the consistency, sensitivity and specificity were 93.4%, 87.2% and 100% respectively compared with the assay using tumor DNA.Conclusion: Patients with CIMP(+) colorectal cancer have unique clinical pathological characteristics and poor prognosis. When tumor tissues are not available, CIMP analysis with plasma free DNA using digital PCR is a feasible alternative.
2015, 22(4):472-478.
DOI: 10.3872/j.issn.1007-385X.2015.04.011
Abstract:
Objective:To investigate the expression and methylation of IGFBP3 gene in esophageal squamous cell carcinoma (ESCC) cell lines and primary tumor tissues, and to explore the relationship between IGFBP3 expression and the clinical pathological features of ESCC. Methods: The mRNA and methylation status of IGFBP3 gene were detected by reverse transcription-PCR (RT-PCR) and methylation specific-PCR (MSP) using RNA and DNA from ESCC cell lines (TE1, TE13, YES-2,T.TN, Eca109) as well as primary tumor tissues and paired normal tissues from 82 ESCC patients. Immunohistochemistry was used to detect the expression of IGFBP3 in ESCC tissues. The relationship among aberrant methylation, expression of IGFBP3 gene, and clinical pathological features were analyzed with the SPSS 13.0 software. Results： IGFBP3 mRNA was undetectable or at very low level in ESCC cell lines examined (TE1, TE13, YES-2,T.TN, Eca109). However, its level increased significantly after the cells were treated with DNA methyltransferase inhibitor, 5-aza-2’-deoxycytidine (5-Aza-dC), indicating that IGFBP3 gene existed is hypermethylated in these cells (P<0.05). In primary tumor tissues from ESCC patients, IGFBP3 mRNA level (0.15±0.07) was significantly lower than that in corresponding normal tissues (0.88±0.32) (P＜0.01). Similarly, the positive rate of IGFBP3 (29.3%, 24/82) in ESCC tissues was significantly lower than that in corresponding normal tissues (84.1%, 69/82) (P<0.01). Thus, The methylation status of IGFBP3 gene associates with its mRNA and protein expression (P<0.05). Moreover, the methylation frequency of IGFBP3 gene in ESCC tissues (68.3%, 56/82) was increased significantly compared to that in corresponding normal tissues (15.9%, 13/82) (P<0.01) and associated with TNM stage of the tumors (P<0.05). The methylation frequency of IGFBP3 gene in stage Ⅲ-Ⅳ tumor tissues was significantly higher than that in stageⅠ-Ⅱtumor tissues (P<0.05). However, the methylation status of IGFBP3 in ESCC tissues was not associated with its pathological grade (P>0.05). Conclusion: The hypermethylation of IGFBP3 gene is a frequent event in ESCC cell lines and primary tumor tissues. The reduced expression of IGFBP3 caused by promoter hypermethylation of the gene may play an important role in the development of ESCC.
Objective:To investigate the expression and methylation of IGFBP3 gene in esophageal squamous cell carcinoma (ESCC) cell lines and primary tumor tissues, and to explore the relationship between IGFBP3 expression and the clinical pathological features of ESCC. Methods: The mRNA and methylation status of IGFBP3 gene were detected by reverse transcription-PCR (RT-PCR) and methylation specific-PCR (MSP) using RNA and DNA from ESCC cell lines (TE1, TE13, YES-2,T.TN, Eca109) as well as primary tumor tissues and paired normal tissues from 82 ESCC patients. Immunohistochemistry was used to detect the expression of IGFBP3 in ESCC tissues. The relationship among aberrant methylation, expression of IGFBP3 gene, and clinical pathological features were analyzed with the SPSS 13.0 software. Results： IGFBP3 mRNA was undetectable or at very low level in ESCC cell lines examined (TE1, TE13, YES-2,T.TN, Eca109). However, its level increased significantly after the cells were treated with DNA methyltransferase inhibitor, 5-aza-2’-deoxycytidine (5-Aza-dC), indicating that IGFBP3 gene existed is hypermethylated in these cells (P<0.05). In primary tumor tissues from ESCC patients, IGFBP3 mRNA level (0.15±0.07) was significantly lower than that in corresponding normal tissues (0.88±0.32) (P＜0.01). Similarly, the positive rate of IGFBP3 (29.3%, 24/82) in ESCC tissues was significantly lower than that in corresponding normal tissues (84.1%, 69/82) (P<0.01). Thus, The methylation status of IGFBP3 gene associates with its mRNA and protein expression (P<0.05). Moreover, the methylation frequency of IGFBP3 gene in ESCC tissues (68.3%, 56/82) was increased significantly compared to that in corresponding normal tissues (15.9%, 13/82) (P<0.01) and associated with TNM stage of the tumors (P<0.05). The methylation frequency of IGFBP3 gene in stage Ⅲ-Ⅳ tumor tissues was significantly higher than that in stageⅠ-Ⅱtumor tissues (P<0.05). However, the methylation status of IGFBP3 in ESCC tissues was not associated with its pathological grade (P>0.05). Conclusion: The hypermethylation of IGFBP3 gene is a frequent event in ESCC cell lines and primary tumor tissues. The reduced expression of IGFBP3 caused by promoter hypermethylation of the gene may play an important role in the development of ESCC.
Peng Junling
,
Tang Tao
,
Ye Zulu
,
Shao Qiong
,
Huang Liyun
,
Deng Ling
,
Wang Fang
,
Shao Jianyong
2015, 22(4):479-483.
DOI: 10.3872/j.issn.1007-385X.2014.5.012
Abstract:
Objective : To investigate the relationship of microsatellite instability (MSI) condition with the deletion of mismatch repair (MMR) proteins (MLH1, MSH2, MSH6, PMS2) and the clinical pathological characteristics in sporadic colorectal cancers. Methods: MSI of DAN samples from sporadic colorectal cancers was examined with multiplex fluorescent PCR. MMR proteins in sporadic colorectal cancer tissues were detected with SP immunohistochemistry (IHC) method. Results: In 75 cases of sporadic colorectal cancers, 21 cases (28%) had MSI, including MSI-H (19 cases) and MSI-L (2). MMR protein deletion was found in 16 cases (21.3%), of which 15 cases (93.75%) were MSI-H, and the other case (6.2%) was MSS. The rate of MMR protein deletion in MSI group (15/21，71.43%) was significantly higher than that of MSS group (1/54，1.9%, P<0.01). Notably, 4 cases with MSI-H (6.78%) and 2 cases (339%) with MSI-L did not have any NMR protein deletion. MSI in these patients was associated with age, mucus content of the adenocarcinoma, and tumor metastasis (P<0.01). MSI-H tends to occur in >50 years old individuals who have mucinous adenocarcinoma harboring MMR protein deletion and often have no distant metastasis of the cancer. Conclusion: The incidence of MSI is higher than that of MMR protein deletion in sporadic colorectal cancer. These with MSI-H have lower risk of metastasis and better prognosis. Therefore, detecting MSI is valuable for improving the prevention, diagnosis, and treatment of sporadic colorectal cancer, and for reducing its incidence and mortality.
Objective : To investigate the relationship of microsatellite instability (MSI) condition with the deletion of mismatch repair (MMR) proteins (MLH1, MSH2, MSH6, PMS2) and the clinical pathological characteristics in sporadic colorectal cancers. Methods: MSI of DAN samples from sporadic colorectal cancers was examined with multiplex fluorescent PCR. MMR proteins in sporadic colorectal cancer tissues were detected with SP immunohistochemistry (IHC) method. Results: In 75 cases of sporadic colorectal cancers, 21 cases (28%) had MSI, including MSI-H (19 cases) and MSI-L (2). MMR protein deletion was found in 16 cases (21.3%), of which 15 cases (93.75%) were MSI-H, and the other case (6.2%) was MSS. The rate of MMR protein deletion in MSI group (15/21，71.43%) was significantly higher than that of MSS group (1/54，1.9%, P<0.01). Notably, 4 cases with MSI-H (6.78%) and 2 cases (339%) with MSI-L did not have any NMR protein deletion. MSI in these patients was associated with age, mucus content of the adenocarcinoma, and tumor metastasis (P<0.01). MSI-H tends to occur in >50 years old individuals who have mucinous adenocarcinoma harboring MMR protein deletion and often have no distant metastasis of the cancer. Conclusion: The incidence of MSI is higher than that of MMR protein deletion in sporadic colorectal cancer. These with MSI-H have lower risk of metastasis and better prognosis. Therefore, detecting MSI is valuable for improving the prevention, diagnosis, and treatment of sporadic colorectal cancer, and for reducing its incidence and mortality.
2015, 22(4):484-488.
DOI: 10.3872/j.issn.1007-385X.2015.04.013
Abstract:
Objective : To investigate the relationship between TMPRSS4 expression in early-stage non-small cell lung cancer and tumor angiogenesis and assess its clinical significance. Methods: Samples were collected from 87 NSCLC patients who had received surgical treatment in the Department of Thoracic Surgery of Qilu Hospital between January 2009 and January 2010. TMPRSS4 expression in cancerous, adjacent and normal tissues was assessed using immunohistochemical staining. Intratumoral microvessel density (MVD) was measured by counting immunostained CD34 positive endothelial cells. For Real-time PCR analysis, 14 matched pairs of tumor tissues and adjacent noncancerous tissues were collected from pulmonary lobectomy specimens of patients with NSCLC immediately after surgery in our department between September and October of 2014. Results: The level of TMPRSS4 mRNA in NSCLC tissues was significantly higher than that of the adjacent lung tissues（1.826±0.453 vs 0.829±0.366, P<0.05). The positive rates of TMPRSS4 in NSCLC tissues and adjacent tissues were significantly increased when compared with that in normal lung tissues（60.9%, 30.0% vs 167%，P<0.05). NSCLC tissues with high TMPRSS4 expression have significantly higher MVD than that of low TMPRSS4-expressing cancers (P<0.05). TMPRSS4 expression and MVD in cancerous tissues of T2 stage was significantly higher than that of T1 cancers（P<0.05). Conclusion: Increased TMPRSS4 expression is associated with high MVD in early-stage NSCLC tissue, and high TMPRSS4 expression and MVD in cancer tissue are closely related to the tumor infiltration depth. Thus, TMPRSS4 is a potential novel target for the treatment of patients with early NSCLC.
Objective : To investigate the relationship between TMPRSS4 expression in early-stage non-small cell lung cancer and tumor angiogenesis and assess its clinical significance. Methods: Samples were collected from 87 NSCLC patients who had received surgical treatment in the Department of Thoracic Surgery of Qilu Hospital between January 2009 and January 2010. TMPRSS4 expression in cancerous, adjacent and normal tissues was assessed using immunohistochemical staining. Intratumoral microvessel density (MVD) was measured by counting immunostained CD34 positive endothelial cells. For Real-time PCR analysis, 14 matched pairs of tumor tissues and adjacent noncancerous tissues were collected from pulmonary lobectomy specimens of patients with NSCLC immediately after surgery in our department between September and October of 2014. Results: The level of TMPRSS4 mRNA in NSCLC tissues was significantly higher than that of the adjacent lung tissues（1.826±0.453 vs 0.829±0.366, P<0.05). The positive rates of TMPRSS4 in NSCLC tissues and adjacent tissues were significantly increased when compared with that in normal lung tissues（60.9%, 30.0% vs 167%，P<0.05). NSCLC tissues with high TMPRSS4 expression have significantly higher MVD than that of low TMPRSS4-expressing cancers (P<0.05). TMPRSS4 expression and MVD in cancerous tissues of T2 stage was significantly higher than that of T1 cancers（P<0.05). Conclusion: Increased TMPRSS4 expression is associated with high MVD in early-stage NSCLC tissue, and high TMPRSS4 expression and MVD in cancer tissue are closely related to the tumor infiltration depth. Thus, TMPRSS4 is a potential novel target for the treatment of patients with early NSCLC.
2015, 22(4):489-494.
DOI: 10.3872/j.issn.1007-385X.2015.04.014
Abstract:
Objective : To investigate the role of Heat-Shock Factor 1 (HSF1) in the development and progression of esophageal squamous cell carcinoma (ESCC). Methods: Cancerous (n=90）and adjacent (n=50) esophageal tissue specimens were collected from 90 patients with ESCC who were surgically treated in Qilu Hospital of Shandong University between January and December of 2009. Immunohistochemical staining was performed to assess the expression of HSF1 in carcinoma and adjacent tissues. Real-time PCR was used to determine the level of HSF1 mRNA in resected ESCC and adjacent tissues from 20 patients enrolled between September and November of 2014. 2 test was carried out to analyze the relationship between HSF1 expression and the clinical and pathological characters of these patients，Kaplan-Meier method was used to calculate their 5-year survival rates，Log-rank test was done to compare survival differences，and Cox regression analysis was utilized to determine the independent prognostic factors. Results: The expression of HSF1 at mRNA or protein levels were significantly elevated in ESCC tissues compared with that in adjacent esophageal tissues (P<0.01). Although it is not associated with the age (P=0.453), gender (P=0.692), smoking history (P=0.318) and drinking history (P=0.367) of these patients statistically, increased HSF1 expression is significantly related to the differentiation stages (P=0.012), lymph node metastasis (P=0.002), and TNM stages of the tumors (P=0.024). Univariate and multivariate analysis indicated that increased expression of HSF1 was associated with tumor relapse and prognosis. Conclusion: HSF1 is highly expressed in ESCC, and the elevated expression is associated with tumor progression and prognosis.
Objective : To investigate the role of Heat-Shock Factor 1 (HSF1) in the development and progression of esophageal squamous cell carcinoma (ESCC). Methods: Cancerous (n=90）and adjacent (n=50) esophageal tissue specimens were collected from 90 patients with ESCC who were surgically treated in Qilu Hospital of Shandong University between January and December of 2009. Immunohistochemical staining was performed to assess the expression of HSF1 in carcinoma and adjacent tissues. Real-time PCR was used to determine the level of HSF1 mRNA in resected ESCC and adjacent tissues from 20 patients enrolled between September and November of 2014. 2 test was carried out to analyze the relationship between HSF1 expression and the clinical and pathological characters of these patients，Kaplan-Meier method was used to calculate their 5-year survival rates，Log-rank test was done to compare survival differences，and Cox regression analysis was utilized to determine the independent prognostic factors. Results: The expression of HSF1 at mRNA or protein levels were significantly elevated in ESCC tissues compared with that in adjacent esophageal tissues (P<0.01). Although it is not associated with the age (P=0.453), gender (P=0.692), smoking history (P=0.318) and drinking history (P=0.367) of these patients statistically, increased HSF1 expression is significantly related to the differentiation stages (P=0.012), lymph node metastasis (P=0.002), and TNM stages of the tumors (P=0.024). Univariate and multivariate analysis indicated that increased expression of HSF1 was associated with tumor relapse and prognosis. Conclusion: HSF1 is highly expressed in ESCC, and the elevated expression is associated with tumor progression and prognosis.
2015, 22(4):495-499.
DOI: 10.3872/j.issn.1007-385X.2015.04.015
Abstract:
Objective : To study the expression and clinical significance of HIF-1α and GLUT-1 in human nasopharyngeal carcinoma (NPC). Methods: The expression of HIF-1α and GLUT-1 were examined in tumor tissues collected from 79 patients with NPC enrolled into our hospital from September 2010 to October 2013 and in nasopharyngitis tissues from 45 patients by using immunohistochemical staining SP method. MVD was counted by immunostaining with anti-CD34 antibody. The expression of HIF-1α, GLUT-1, and VEFG were also determined in CNE1 cells cultured under hypoxia condition. Results: Immunohistochemical staining showed that HIF-1α was highly expressed in nuclei of NPC (positive rate 63.40%) but not in nasopharyngitis tissues. The level of HIF-1α expression was associated with tumor size，lymph metastasis, T stage and clinic stage (P<0.05). HIF-1α expression was also positively correlated with MVD, which was significant higher in NPC. GLUT-1 was weakly expressed in nasopharyngitis tissues (positive rate 6.67%) but strongly expressed in NPC tissues (positive rate 53.16%). The expression of GLUT-1 in NPC was associated with MVD, lymph metastasis and clinic stage (P<0.05). Additionally, hypoxia increased the levels of HIF-1a, GLUT-1 and VEGF in CNE1 cells. Conclusion: The expression of HIF-1α and GLUT-1 are elevated in NPC, which are associated with pathological features and increased MVD. The in vitro study supported the notion that HIF-1α promotes the growth of nasopharyngeal cancer blood vessels through activating VEGF production. These results also suggest that HIF-1a and GLUT-1 are potential targets for the diagnosis and treatment of NPC.
Objective : To study the expression and clinical significance of HIF-1α and GLUT-1 in human nasopharyngeal carcinoma (NPC). Methods: The expression of HIF-1α and GLUT-1 were examined in tumor tissues collected from 79 patients with NPC enrolled into our hospital from September 2010 to October 2013 and in nasopharyngitis tissues from 45 patients by using immunohistochemical staining SP method. MVD was counted by immunostaining with anti-CD34 antibody. The expression of HIF-1α, GLUT-1, and VEFG were also determined in CNE1 cells cultured under hypoxia condition. Results: Immunohistochemical staining showed that HIF-1α was highly expressed in nuclei of NPC (positive rate 63.40%) but not in nasopharyngitis tissues. The level of HIF-1α expression was associated with tumor size，lymph metastasis, T stage and clinic stage (P<0.05). HIF-1α expression was also positively correlated with MVD, which was significant higher in NPC. GLUT-1 was weakly expressed in nasopharyngitis tissues (positive rate 6.67%) but strongly expressed in NPC tissues (positive rate 53.16%). The expression of GLUT-1 in NPC was associated with MVD, lymph metastasis and clinic stage (P<0.05). Additionally, hypoxia increased the levels of HIF-1a, GLUT-1 and VEGF in CNE1 cells. Conclusion: The expression of HIF-1α and GLUT-1 are elevated in NPC, which are associated with pathological features and increased MVD. The in vitro study supported the notion that HIF-1α promotes the growth of nasopharyngeal cancer blood vessels through activating VEGF production. These results also suggest that HIF-1a and GLUT-1 are potential targets for the diagnosis and treatment of NPC.
2015, 22(4):500-503.
DOI: 10.3872/j.issn.1007-385X.2015.04.016
Abstract:
Objective : To investigate the clinical pathological features of patients with non-small cell lung cancer who had anti-Hu antibody and whether the presence of the antibody is associated with distant metastasis of the tumor. Methods: Peripheral blood samples were collected from 75 NSCLC patients enrolled into the Department of Oncology between June and December of 2013, and from 100 healthy volunteers who did not have tumor or neurologic history and underwent blood test between November and December of 2013 in the Clinical laboratory, General Hospital, Jinan Command of the People’s Liberation Army. Anti-Hu antibody was measured by ELISA. Clinical pathological features investigated include sex, age, pathological type, differentiation stage, EGFR mutation, tumor markers (CEA, CY21-1, and NSE), and distant metastasis. Results: Compared with healthy volunteers, NSCLC patients had significantly elevated anti-Hu antibody titration (40.00±35.76 ng/ml vs 16.40±8.19 ng/ml, P<0.01). The increase of the anti-Hu antibody was significantly associated with brain metastasis (81.81% vs 50.00%, P=0.015), but not distant metastasis to other tissues. It was also independent of sex, age, gender, pathological stage, EGFR mutation, and the levels of CEA and NSE (all P>005). Multivariate regression analysis further indicated that increased anti-Hu antibody level was an independent variable for brain metastasis prediction (P=0.015). Conclusion: Peripheral blood anti-Hu antibody is increased in NSCLC patients and is associated with brain metastasis of the tumor, which deserves further investigation to understand the mechanism.
Objective : To investigate the clinical pathological features of patients with non-small cell lung cancer who had anti-Hu antibody and whether the presence of the antibody is associated with distant metastasis of the tumor. Methods: Peripheral blood samples were collected from 75 NSCLC patients enrolled into the Department of Oncology between June and December of 2013, and from 100 healthy volunteers who did not have tumor or neurologic history and underwent blood test between November and December of 2013 in the Clinical laboratory, General Hospital, Jinan Command of the People’s Liberation Army. Anti-Hu antibody was measured by ELISA. Clinical pathological features investigated include sex, age, pathological type, differentiation stage, EGFR mutation, tumor markers (CEA, CY21-1, and NSE), and distant metastasis. Results: Compared with healthy volunteers, NSCLC patients had significantly elevated anti-Hu antibody titration (40.00±35.76 ng/ml vs 16.40±8.19 ng/ml, P<0.01). The increase of the anti-Hu antibody was significantly associated with brain metastasis (81.81% vs 50.00%, P=0.015), but not distant metastasis to other tissues. It was also independent of sex, age, gender, pathological stage, EGFR mutation, and the levels of CEA and NSE (all P>005). Multivariate regression analysis further indicated that increased anti-Hu antibody level was an independent variable for brain metastasis prediction (P=0.015). Conclusion: Peripheral blood anti-Hu antibody is increased in NSCLC patients and is associated with brain metastasis of the tumor, which deserves further investigation to understand the mechanism.
2015, 22(4):505-508.
DOI: 10.3872/j.issn.1007-385X.2015.04.017
Abstract:
Objective : The aim of the present study is to evaluate the effects of autologous cytokine-induced killer cells (CIK) immunotherapy on immune function and quality of life of cancer patients. Methods: We performed a retrospective analysis of 58 cancer patients who received CIK immunotherapy in our hospital. The T lymphocyte in their peripheral blood samples drew before and after the treatment were analyzed by flow cytometry for CD3+, CD3+CD4+, CD3+CD8+,CD3-CD16+CD56+ and CD3+CD16+CD56+ subpopulations. Patients were assessed with the WHO QOL-BREF questionnaires to evaluate their quality of life. Treatment-related adverse reactions during treatment were also surveyed. Results: In the culture for generating CIK, the levels of CD3+ T lymphocytes, CD3+CD4+ T helper cells, CD3+CD8+ cytotoxic T cells were (82.79±11.98)%, (30.32±11.23)%, and (49.10±11.65)% respectively. The ratio of CD4+ and CD8+ T cells was 0.67±0.34. Notably, the levels of CD3+CD16+CD56+ CIK cells and CD3-CD16+CD56+ NK cells were (16.58±11.83)% and (13.74±8.66)%. Comparing the two peripheral blood samples from each patient, the percentages of CD3+ T lymphocytes, CD3+CD4+ T helper cells, CD3+CD8+ cytotoxic T cells, and CD3+CD16+CD56+CIK subsets were all significantly increased after the treatment (P<0.05), whereas the percentage of CD3-CD16+CD56+NK cell percentage decreased significantly (P<0.05). The QOL-BREF of patients were also improved significantly (P<005), and the treatment-related adverse reactions were well-tolerated in most patients. Conclusion: CIK therapy is safe and effective in improving quality of life and immune functions of patients with malignant tumors.
Objective : The aim of the present study is to evaluate the effects of autologous cytokine-induced killer cells (CIK) immunotherapy on immune function and quality of life of cancer patients. Methods: We performed a retrospective analysis of 58 cancer patients who received CIK immunotherapy in our hospital. The T lymphocyte in their peripheral blood samples drew before and after the treatment were analyzed by flow cytometry for CD3+, CD3+CD4+, CD3+CD8+,CD3-CD16+CD56+ and CD3+CD16+CD56+ subpopulations. Patients were assessed with the WHO QOL-BREF questionnaires to evaluate their quality of life. Treatment-related adverse reactions during treatment were also surveyed. Results: In the culture for generating CIK, the levels of CD3+ T lymphocytes, CD3+CD4+ T helper cells, CD3+CD8+ cytotoxic T cells were (82.79±11.98)%, (30.32±11.23)%, and (49.10±11.65)% respectively. The ratio of CD4+ and CD8+ T cells was 0.67±0.34. Notably, the levels of CD3+CD16+CD56+ CIK cells and CD3-CD16+CD56+ NK cells were (16.58±11.83)% and (13.74±8.66)%. Comparing the two peripheral blood samples from each patient, the percentages of CD3+ T lymphocytes, CD3+CD4+ T helper cells, CD3+CD8+ cytotoxic T cells, and CD3+CD16+CD56+CIK subsets were all significantly increased after the treatment (P<0.05), whereas the percentage of CD3-CD16+CD56+NK cell percentage decreased significantly (P<0.05). The QOL-BREF of patients were also improved significantly (P<005), and the treatment-related adverse reactions were well-tolerated in most patients. Conclusion: CIK therapy is safe and effective in improving quality of life and immune functions of patients with malignant tumors.
2015, 22(4):509-513.
DOI: 10.3872/j.issn.1007-385X.2015.04.018
Abstract:
Objective : To evaluate the clinical efficacy of combined therapy using autologous dendritic cells-cytokine induced killer cells (DC-CIK) and percutaneous microwave coagulation (PMCT) in the treatment of hepatocellular carcinoma. Methods: Forty four patients with hepatocellular carcinoma treated with PMCT and DC-CIK (combination group) and 44 patients treated with PMCT alone (control group) were enrolled in the Department of Biotherapy of Eastern Hepatobiliary Surgical Hospital from January 2012 to February 2014. For patients in the combination group, peripheral blood mononuclear cells (PBMCs) were isolated on day 1 and PMCT were performed on the second day. After a ten day-culture, the DC-CIK cells were generated and transfused back to patients together with intradermal injection of DC vaccine. The second and third course of DC-CIK therapy were given one month and two months later respectively. The AFP level, immune function, progression-free survival, overall survival, and adverse effects were assessed for all the patients. Results: Although the level of AFP and regulatory T cells in peripheral blood decreased in the patients of both groups, their decline in the combination group was significantly more than that of the control group (P<0.05). Compared with that of pre-treatment, the lymphocyte number and subpopulations didn’t change significantly in the control group (P>0.05), but they did increase markedly in the combination group (P<0.05). The median progression-free survival and median overall survival were both increased in the combination group compared with that of the control group (7.1 m vs 4.9 m, 215 m vs 14.0 m, P<0.05). Conclusion: Adoptive transfer of autologous DC-CIK in combination with PMCT is an effective treatment for patients with hepatocellular carcinoma. It improves the immune function, postpones the recurrence of tumor, and prolongs the overall survival with acceptable toxicities.
Objective : To evaluate the clinical efficacy of combined therapy using autologous dendritic cells-cytokine induced killer cells (DC-CIK) and percutaneous microwave coagulation (PMCT) in the treatment of hepatocellular carcinoma. Methods: Forty four patients with hepatocellular carcinoma treated with PMCT and DC-CIK (combination group) and 44 patients treated with PMCT alone (control group) were enrolled in the Department of Biotherapy of Eastern Hepatobiliary Surgical Hospital from January 2012 to February 2014. For patients in the combination group, peripheral blood mononuclear cells (PBMCs) were isolated on day 1 and PMCT were performed on the second day. After a ten day-culture, the DC-CIK cells were generated and transfused back to patients together with intradermal injection of DC vaccine. The second and third course of DC-CIK therapy were given one month and two months later respectively. The AFP level, immune function, progression-free survival, overall survival, and adverse effects were assessed for all the patients. Results: Although the level of AFP and regulatory T cells in peripheral blood decreased in the patients of both groups, their decline in the combination group was significantly more than that of the control group (P<0.05). Compared with that of pre-treatment, the lymphocyte number and subpopulations didn’t change significantly in the control group (P>0.05), but they did increase markedly in the combination group (P<0.05). The median progression-free survival and median overall survival were both increased in the combination group compared with that of the control group (7.1 m vs 4.9 m, 215 m vs 14.0 m, P<0.05). Conclusion: Adoptive transfer of autologous DC-CIK in combination with PMCT is an effective treatment for patients with hepatocellular carcinoma. It improves the immune function, postpones the recurrence of tumor, and prolongs the overall survival with acceptable toxicities.
2015, 22(4):514-518.
DOI: 10.3872/j.issn.1007-385X.2015.04.019
Abstract:
胃癌是恶性肿瘤中的常见病种，其肿瘤相关的病死率占全世界第三位，经传统手术及放化疗治疗后胃癌的生存率仍然很低。生物免疫治疗中肿瘤疫苗的研究在近几年取得了重大进步，根据肿瘤患者的个体遗传基因结构或蛋白表达的不同，选择个体化的肿瘤疫苗可以进一步提高治疗效果。个体化肿瘤抗原高表达肽疫苗、负载的树突状细胞疫苗以及个体化多肽疫苗等已在胃癌免疫治疗的临床转化研究中取得了很多成果，个体化肽疫苗与放化疗、分子靶向治疗等其他治疗联合应用可以有效提高临床疗效。本文介绍个体化肽疫苗在胃癌领域的临床转化研究进展、其与化疗、放疗等多学科结合的综合治疗模式及其相关疗效评价等免疫治疗临床转化的新思路和新方法。
胃癌是恶性肿瘤中的常见病种，其肿瘤相关的病死率占全世界第三位，经传统手术及放化疗治疗后胃癌的生存率仍然很低。生物免疫治疗中肿瘤疫苗的研究在近几年取得了重大进步，根据肿瘤患者的个体遗传基因结构或蛋白表达的不同，选择个体化的肿瘤疫苗可以进一步提高治疗效果。个体化肿瘤抗原高表达肽疫苗、负载的树突状细胞疫苗以及个体化多肽疫苗等已在胃癌免疫治疗的临床转化研究中取得了很多成果，个体化肽疫苗与放化疗、分子靶向治疗等其他治疗联合应用可以有效提高临床疗效。本文介绍个体化肽疫苗在胃癌领域的临床转化研究进展、其与化疗、放疗等多学科结合的综合治疗模式及其相关疗效评价等免疫治疗临床转化的新思路和新方法。
20 The role of STAT3 in the immune tolerance and the radiation and chemotherapy insensitivity of tumor
2015, 22(4):519-523.
DOI: 10.3872/j.issn.1007-385X.2015.04.020
Abstract:
信号转导和转录激活因子3 (signal transducer and activator of transcription 3，STAT3)是近年来研究肿瘤细胞内异常活化的重要转录因子之一，由多种细胞因子和生长因子等多肽类配体激活。在肿瘤微环境中，STAT3呈持续性活化状态，不仅造成肿瘤对放、化疗的不敏感，降低了临床疗效，而且也参与了肿瘤免疫耐受现象的形成，削弱了免疫治疗的效果。因此，STAT3有可能成为克服肿瘤治疗障碍的一个理想靶点。
信号转导和转录激活因子3 (signal transducer and activator of transcription 3，STAT3)是近年来研究肿瘤细胞内异常活化的重要转录因子之一，由多种细胞因子和生长因子等多肽类配体激活。在肿瘤微环境中，STAT3呈持续性活化状态，不仅造成肿瘤对放、化疗的不敏感，降低了临床疗效，而且也参与了肿瘤免疫耐受现象的形成，削弱了免疫治疗的效果。因此，STAT3有可能成为克服肿瘤治疗障碍的一个理想靶点。
2015, 22(4):524-527.
DOI: 10.3872/j.issn.1007-385X.2015.04.021
Abstract:
整合素作为细胞表面受体、细胞间的黏附分子，对细胞与细胞间、细胞与细胞外基质间的黏附起介导作用。近几十年来，恶性肿瘤甲状腺癌的发病率急速攀升，很大原因是由于恶性肿瘤细胞极易增殖、侵袭和转移，而这种特性与细胞表面黏附分子整合素有密切的关系。有研究发现，整合素α3β1、α6β1、αvβ3等通过与细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)、血小板反应蛋白(thrombospondin, TSP)和细胞外基质胶原(collagen, CO)、层黏连蛋白(laminin, LN)、纤连蛋白(fibronectin, FN)等配体结合，激活FAK-Ras-Raf或磷脂酰肌醇3激酶(PI3K)-Akt-mTOR等信号转导通路，调控甲状腺癌细胞的生长、繁殖、侵袭和转移。通过针对整合素信号通路上多个靶点进行分子靶向干预，可能研发出甲状腺癌的针对性治疗方案。本文主要对整合素在甲状腺癌的增殖、侵袭、转移、诊断和治疗等方面所起的作用作一综述。
整合素作为细胞表面受体、细胞间的黏附分子，对细胞与细胞间、细胞与细胞外基质间的黏附起介导作用。近几十年来，恶性肿瘤甲状腺癌的发病率急速攀升，很大原因是由于恶性肿瘤细胞极易增殖、侵袭和转移，而这种特性与细胞表面黏附分子整合素有密切的关系。有研究发现，整合素α3β1、α6β1、αvβ3等通过与细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)、血小板反应蛋白(thrombospondin, TSP)和细胞外基质胶原(collagen, CO)、层黏连蛋白(laminin, LN)、纤连蛋白(fibronectin, FN)等配体结合，激活FAK-Ras-Raf或磷脂酰肌醇3激酶(PI3K)-Akt-mTOR等信号转导通路，调控甲状腺癌细胞的生长、繁殖、侵袭和转移。通过针对整合素信号通路上多个靶点进行分子靶向干预，可能研发出甲状腺癌的针对性治疗方案。本文主要对整合素在甲状腺癌的增殖、侵袭、转移、诊断和治疗等方面所起的作用作一综述。
2015, 22(4):528-535.
DOI: 10.3872/j.issn.1007-385X.2015.04.022
Abstract:
信号素（semaphorin，Sema）最初被认定为是参与神经系统发育的轴突信号因子。研究表明，信号素参与生理或病理性免疫反应的不同阶段，能够调节免疫细胞（CD4+ T细胞、B细胞）的活化或分化，促进DC的迁移。 其中，Plexin家庭成员是信号素分子最具代表性的受体，目前已经解析了Sema-Plexin复合体的晶体结构，为后续开展肿瘤的免疫治疗提供了依据。在本文中，我们综述了信号素家族分子及其受体的结构基础，以及各个信号素分子在免疫细胞（T细胞、B细胞以及DC）功能的最新研究进展。
信号素（semaphorin，Sema）最初被认定为是参与神经系统发育的轴突信号因子。研究表明，信号素参与生理或病理性免疫反应的不同阶段，能够调节免疫细胞（CD4+ T细胞、B细胞）的活化或分化，促进DC的迁移。 其中，Plexin家庭成员是信号素分子最具代表性的受体，目前已经解析了Sema-Plexin复合体的晶体结构，为后续开展肿瘤的免疫治疗提供了依据。在本文中，我们综述了信号素家族分子及其受体的结构基础，以及各个信号素分子在免疫细胞（T细胞、B细胞以及DC）功能的最新研究进展。
2015, 22(4):536-540.
DOI: 10.3872/j.issn.1007-385X.2015.04.023
Abstract:
神经菌毛素（neuropilin-1，Nrp-1）是一种多功能的非酪氨酸激酶受体，在内皮细胞及多种肿瘤细胞表面高表达。Nrp-1主要作为血管内皮细胞生长因子165（vascular endothelial growth factor 165，VEGF165）的共受体，可显著提高VEGF165受体（vascular endothelial growth factor receptor 2，KDR）与VEGF165结合，促进肿瘤血管新生及肿瘤迁移。近期的研究表明，Nrp-1还能通过与脑信号蛋白Ⅲ（semaphorin Ⅲ，SEMA3）相互作用、与转化生长因子β（transforming growth factor-β，TGF-β）相互作用从而促进纤连蛋白聚集，改善肿瘤微环境，并介导调节性T细胞到肿瘤组织等多种途径，促进肿瘤的发生发展。由于Nrp-1在于肿瘤发生发展中所起到的病理性促进作用，Nrp-1逐渐成为肿瘤治疗的一个新靶标，本文主要介绍Nrp-1的生物学特征以及其对于肿瘤发生发展中的信号转导作用。
神经菌毛素（neuropilin-1，Nrp-1）是一种多功能的非酪氨酸激酶受体，在内皮细胞及多种肿瘤细胞表面高表达。Nrp-1主要作为血管内皮细胞生长因子165（vascular endothelial growth factor 165，VEGF165）的共受体，可显著提高VEGF165受体（vascular endothelial growth factor receptor 2，KDR）与VEGF165结合，促进肿瘤血管新生及肿瘤迁移。近期的研究表明，Nrp-1还能通过与脑信号蛋白Ⅲ（semaphorin Ⅲ，SEMA3）相互作用、与转化生长因子β（transforming growth factor-β，TGF-β）相互作用从而促进纤连蛋白聚集，改善肿瘤微环境，并介导调节性T细胞到肿瘤组织等多种途径，促进肿瘤的发生发展。由于Nrp-1在于肿瘤发生发展中所起到的病理性促进作用，Nrp-1逐渐成为肿瘤治疗的一个新靶标，本文主要介绍Nrp-1的生物学特征以及其对于肿瘤发生发展中的信号转导作用。
2015, 22(4):541-544.
DOI: 10.3872/j.issn.1007-385X.2015.04.024
Abstract:
肺腺癌转移相关转录1（metastasis-associated lung adenocarcinoma，MALAT-1）自首次发现以来，其在肿瘤中作用逐渐成为非编码长链RNA（long non-coding RNA，lncRNA）研究中的热点。MALAT-1主要通过调控可变剪接及直接调控编码蛋白质基因表达，参与肿瘤相关基因的表观遗传调控及相关信号转导通路，影响肿瘤生长、侵袭及转移、抗凋亡、耐药形成。本文就MALAT-1结构、生物学功能特点及其在肿瘤发生、发展中的作用和机制进行简述。
肺腺癌转移相关转录1（metastasis-associated lung adenocarcinoma，MALAT-1）自首次发现以来，其在肿瘤中作用逐渐成为非编码长链RNA（long non-coding RNA，lncRNA）研究中的热点。MALAT-1主要通过调控可变剪接及直接调控编码蛋白质基因表达，参与肿瘤相关基因的表观遗传调控及相关信号转导通路，影响肿瘤生长、侵袭及转移、抗凋亡、耐药形成。本文就MALAT-1结构、生物学功能特点及其在肿瘤发生、发展中的作用和机制进行简述。
2015, 22(4):545-546.
DOI: 10.3872/j.issn.1007-385X.2015.04.025
Abstract:
毛细血管渗漏综合征（capillary leak syndrome, CLS）是以低血压、低蛋白血症和全身水肿为主要表现的临床综合征，CLS患者通常病情危重，严重时可致心、肺及肾等多器官功能衰竭，病死率较高\[1\]。引起CLS的病因较多，而自体细胞因子诱导的杀伤（cytokine-induced killer，CIK）细胞回输导致CLS，国内鲜有报道。为提高对该严重并发症的认识，现将我科诊治的1 例晚期乳腺癌患者经自体CIK细胞治疗后出现CLS 的诊治过程报告如下。
毛细血管渗漏综合征（capillary leak syndrome, CLS）是以低血压、低蛋白血症和全身水肿为主要表现的临床综合征，CLS患者通常病情危重，严重时可致心、肺及肾等多器官功能衰竭，病死率较高\[1\]。引起CLS的病因较多，而自体细胞因子诱导的杀伤（cytokine-induced killer，CIK）细胞回输导致CLS，国内鲜有报道。为提高对该严重并发症的认识，现将我科诊治的1 例晚期乳腺癌患者经自体CIK细胞治疗后出现CLS 的诊治过程报告如下。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.