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Volume 18,Issue 5,2011 Table of Contents
2011, 18(5):467-472.
DOI: 10.3872/j.issn.1007-385X.2011.5.001
Abstract:
With the advances in molecular biology and the clarification of antibody genetic structure, a variety of smallmolecule antibody basedand genetic manipulatedgenetic engineering antibody fragments have emerged and been widely used in scientific research, clinical diagnosis and therapy. Among which, bispecific antibody with combined biological activity and therapeutic targeting activity, immunoconjugate with enhanced function and targeting cytotoxicity, nanobody with special single domain structure which is widely applied in biological technology and clinical research, and intrabody with special intracellular expression are the most active fields in antibody fragment research. Genetic engineering antibody fragments have received extensive attention and approval over fulllength monoclonal antibodies owing to their desirable characteristics, such as weak immunogenicity, strong tissue permeability, minimal toxicity and simple preparation. although the safety and efficiency of most antibody fragments in clinical research still need to be further identified, development of scientific research on genetic engineering antibody fragments, will inevitably improve diagnosis and therapy of human diseases.
With the advances in molecular biology and the clarification of antibody genetic structure, a variety of smallmolecule antibody basedand genetic manipulatedgenetic engineering antibody fragments have emerged and been widely used in scientific research, clinical diagnosis and therapy. Among which, bispecific antibody with combined biological activity and therapeutic targeting activity, immunoconjugate with enhanced function and targeting cytotoxicity, nanobody with special single domain structure which is widely applied in biological technology and clinical research, and intrabody with special intracellular expression are the most active fields in antibody fragment research. Genetic engineering antibody fragments have received extensive attention and approval over fulllength monoclonal antibodies owing to their desirable characteristics, such as weak immunogenicity, strong tissue permeability, minimal toxicity and simple preparation. although the safety and efficiency of most antibody fragments in clinical research still need to be further identified, development of scientific research on genetic engineering antibody fragments, will inevitably improve diagnosis and therapy of human diseases.
DU Chunjuan
,
YU Jinpu
,
LI Hui
,
ZHANG Naining
,
TENG Yanhua
,
LI Zhen
,
AN Yang
,
CAO Shui
,
REN Xiubao
2011, 18(5):473-479.
DOI: 10.3872/j.issn.1007-385X.2011.5.002
Abstract:
Objective:To observe the alteration in proportion of CD4+CD25+ regulatory T cells (Tregs) in the peripheral blood of lung cancer patients treated with GMCSF modified tumor cell vaccine (GVAX) and its relationship to clinical characteristics and survival rate of lung cancer patients. Methods: Eightyfive lung cancer patients treated with GVAX in Affiliated Tumor Hospital of Tianjin Medical University were enrolled in the present study, including 28 squamous carcinoma, 47 adenocarcinoma, 10 small cell lung cancers, among which 23 in early stage (Ⅰ~ⅢA), 62 in advanced stage (ⅢB~Ⅳ). The proportion of Tregs in the peripheral blood before and after GVAX treatment was detected by flow cytometry, and the correlation between the alteration of Treg proportional and clinicopathological characteristics and overall survival of lung cancer patients were analyzed.Results: Oneyear, 3year survival rates of 85 patients with lung cancer treated with GVAX were 69.3% and 48.2%, respectively. The proportion of Treg in peripheral blood significantly decreased after GVAX treatment (\[4.86±2.52\]% vs \[5.52±2.68\]%, P<0.05). The proportion of Tregs significantly decreased in the patients with normal LDH level (\[4.50±2.23\]% vs \[5.59±2.76\]%, P<0.05) while increased in those with high LDH level (\[6.00±3.06\]% vs \[5.30±2.47\]%, P<0.05) after GVAX treatment. The proportion of Tregs decreased more dramatically in the patients treated with GVAX more than 1 course than in those patients with only 1 course (\[-2.11±1.620\]% vs \[-0.39±2.39\]%, P<0.05). The alteration of Treg proportion after GVAX treatment was tightly related to the prognosis of the patients. Patients with a decrease of Tregs showed longer overall survival (OS) than those displaying an increase of Treg after GVAX treatment (21 months vs 10 months, P<0.05), especially patients in advanced stage (18 months vs 8 months, P<0.05). Conclusion: The alteration of Treg proportion in peripheral blood could be a potential immunological indicator to evaluate the efficacy and prognosis of lung cancer patients who received GVAX treatment. The declining pattern of Treg after GVAX treatment implies a better prognosis of patients.
Objective:To observe the alteration in proportion of CD4+CD25+ regulatory T cells (Tregs) in the peripheral blood of lung cancer patients treated with GMCSF modified tumor cell vaccine (GVAX) and its relationship to clinical characteristics and survival rate of lung cancer patients. Methods: Eightyfive lung cancer patients treated with GVAX in Affiliated Tumor Hospital of Tianjin Medical University were enrolled in the present study, including 28 squamous carcinoma, 47 adenocarcinoma, 10 small cell lung cancers, among which 23 in early stage (Ⅰ~ⅢA), 62 in advanced stage (ⅢB~Ⅳ). The proportion of Tregs in the peripheral blood before and after GVAX treatment was detected by flow cytometry, and the correlation between the alteration of Treg proportional and clinicopathological characteristics and overall survival of lung cancer patients were analyzed.Results: Oneyear, 3year survival rates of 85 patients with lung cancer treated with GVAX were 69.3% and 48.2%, respectively. The proportion of Treg in peripheral blood significantly decreased after GVAX treatment (\[4.86±2.52\]% vs \[5.52±2.68\]%, P<0.05). The proportion of Tregs significantly decreased in the patients with normal LDH level (\[4.50±2.23\]% vs \[5.59±2.76\]%, P<0.05) while increased in those with high LDH level (\[6.00±3.06\]% vs \[5.30±2.47\]%, P<0.05) after GVAX treatment. The proportion of Tregs decreased more dramatically in the patients treated with GVAX more than 1 course than in those patients with only 1 course (\[-2.11±1.620\]% vs \[-0.39±2.39\]%, P<0.05). The alteration of Treg proportion after GVAX treatment was tightly related to the prognosis of the patients. Patients with a decrease of Tregs showed longer overall survival (OS) than those displaying an increase of Treg after GVAX treatment (21 months vs 10 months, P<0.05), especially patients in advanced stage (18 months vs 8 months, P<0.05). Conclusion: The alteration of Treg proportion in peripheral blood could be a potential immunological indicator to evaluate the efficacy and prognosis of lung cancer patients who received GVAX treatment. The declining pattern of Treg after GVAX treatment implies a better prognosis of patients.
2011, 18(5):480-484.
DOI: 10.3872/j.issn.1007-385X.2011.5.003
Abstract:
Objective:To evaluate the efficacy of cytokineinduced killer cells（CIK）in treatment of patients with renal cell carcinoma. Methods: All the patients were diagnosed as renal cell carcinoma in Affiliated Cancer Hospital of Tianjin Medical University from January 2000 to July 2010. One hundred and nineteen patients received CIK treatment (CIK group) and 119 patients received treatment of IL2 in combination with IFN (control group). Of the 119 paired patients, 21 pairs had stage Ⅰ disease, 21 pairs stage Ⅱ, 49 stage Ⅲ, and 28 stage Ⅳ. Pairing consideration include clinical stage,sex,age,neutrophil count, platelet count,hemoglobin level,lactate dehydrogenase activity,β2microglobulin level and Karnofsky performance status at the time of diagnosis. Progressionfree survival (PFS) and overall survival (OS) were evaluated. Results:The 5year PFS and OS rates in the CIK and control groups were 44% and 42% (P=0.056), and 72% and 51% (P<0.001), respectively. The median PFS and OS in the CIK and control groups were 54 and 43 months (P=0.088), and 134 and 60 months (P<0.001), respectively. Patients with stage Ⅰ+Ⅱ disease in these two groups showed no statistical difference in PFS and OS. However, the 5year PFS and OS of stage Ⅲ+Ⅳ patients in the CIK group were significantly higher than those in the control group (26% vs 18%, P<0.001, and 58% vs 31%, P<0.001; respectively), the median PFS and OS of stage Ⅲ+Ⅳ patients in the CIK group were also remarkably longer than those in the control group (36 months vs 13 months, P=0.005; and 68 months vs 33 months, P<0.001; respectively). In the multivariate analysis, the frequency of CIK immunotherapy was related to the PFS (HR=0.95, 95% CI: 0.92-0.99, P=0.013) and OS (HR=0.79, 95% CI: 0.71-0.87, P<0.001). The optimal outpoint of the frequency was seven times. Conclusion: CIK immunotherapy can improve the prognosis of stage Ⅲ and Ⅳ renal cell carcinoma patients and increasing the frequency of CIK treatment benefits patients more.
Objective:To evaluate the efficacy of cytokineinduced killer cells（CIK）in treatment of patients with renal cell carcinoma. Methods: All the patients were diagnosed as renal cell carcinoma in Affiliated Cancer Hospital of Tianjin Medical University from January 2000 to July 2010. One hundred and nineteen patients received CIK treatment (CIK group) and 119 patients received treatment of IL2 in combination with IFN (control group). Of the 119 paired patients, 21 pairs had stage Ⅰ disease, 21 pairs stage Ⅱ, 49 stage Ⅲ, and 28 stage Ⅳ. Pairing consideration include clinical stage,sex,age,neutrophil count, platelet count,hemoglobin level,lactate dehydrogenase activity,β2microglobulin level and Karnofsky performance status at the time of diagnosis. Progressionfree survival (PFS) and overall survival (OS) were evaluated. Results:The 5year PFS and OS rates in the CIK and control groups were 44% and 42% (P=0.056), and 72% and 51% (P<0.001), respectively. The median PFS and OS in the CIK and control groups were 54 and 43 months (P=0.088), and 134 and 60 months (P<0.001), respectively. Patients with stage Ⅰ+Ⅱ disease in these two groups showed no statistical difference in PFS and OS. However, the 5year PFS and OS of stage Ⅲ+Ⅳ patients in the CIK group were significantly higher than those in the control group (26% vs 18%, P<0.001, and 58% vs 31%, P<0.001; respectively), the median PFS and OS of stage Ⅲ+Ⅳ patients in the CIK group were also remarkably longer than those in the control group (36 months vs 13 months, P=0.005; and 68 months vs 33 months, P<0.001; respectively). In the multivariate analysis, the frequency of CIK immunotherapy was related to the PFS (HR=0.95, 95% CI: 0.92-0.99, P=0.013) and OS (HR=0.79, 95% CI: 0.71-0.87, P<0.001). The optimal outpoint of the frequency was seven times. Conclusion: CIK immunotherapy can improve the prognosis of stage Ⅲ and Ⅳ renal cell carcinoma patients and increasing the frequency of CIK treatment benefits patients more.
2011, 18(5):485-489.
DOI: 10.3872/j.issn.1007-385X.2011.5.004
Abstract:
Objective:To investigate the effect of hsamiR132 (miR132) on proliferation and invasion of human esophageal carcinoma cells, and further explore its possible mechanism. Methods: pCDNA3.1(+)miR132 plasmid was constructed, and KYSE150 cells stablely overexpressing miR132 were selected through G418 screening. Soft agar colony formation test and Transwell assay were performed to analyze the proliferation and invasion of KYSE150 cells. Expression of miR132 and forkhead box protens A1 (FOXA1) in KYSE150 cells were examined by realtime PCR and Western blotting. The regulatory effect of miR132 overexpression on FOXA1 was further studied by reporter gene assay and Western blotting. Results: pCDNA3.1(+)miR132 plasmid was successfully constructed, and KYSE150 cells stablely overexpressing miR132 was established. In parental KYSE150 cells, miR132 expression was low ,while FOXA1 expression was high. Compared to mock transfected KYSE150 cells and untransfected KYSE150 cells, pCDNA3.1(+)miR132 transfection did not affect the proliferation of KYSE150 cells (13.9±0.33 vs 15.4±0.11, 17.1±0.20, P>0.05), however tumor size reduced and tumor appeared smoother after pCDNA3.1(+)miR132 transfection. Furthermore, overexpression of miR132 significantly suppressed the invasion of KYSE150 cells（55±1.6 vs 129.0±3.1, 124.0±2.8, P＜001), and miR132 decreased the expression of endogenous FOXA1 by targeting FOXA1 3'UTR. Conclusion: Esophageal carcinoma KYSE150 cells express low level of miR132, and overexpression of miR132 can negatively regulate the expression of FOXA1 and suppressed the invasion of KYSE150 cells.
Objective:To investigate the effect of hsamiR132 (miR132) on proliferation and invasion of human esophageal carcinoma cells, and further explore its possible mechanism. Methods: pCDNA3.1(+)miR132 plasmid was constructed, and KYSE150 cells stablely overexpressing miR132 were selected through G418 screening. Soft agar colony formation test and Transwell assay were performed to analyze the proliferation and invasion of KYSE150 cells. Expression of miR132 and forkhead box protens A1 (FOXA1) in KYSE150 cells were examined by realtime PCR and Western blotting. The regulatory effect of miR132 overexpression on FOXA1 was further studied by reporter gene assay and Western blotting. Results: pCDNA3.1(+)miR132 plasmid was successfully constructed, and KYSE150 cells stablely overexpressing miR132 was established. In parental KYSE150 cells, miR132 expression was low ,while FOXA1 expression was high. Compared to mock transfected KYSE150 cells and untransfected KYSE150 cells, pCDNA3.1(+)miR132 transfection did not affect the proliferation of KYSE150 cells (13.9±0.33 vs 15.4±0.11, 17.1±0.20, P>0.05), however tumor size reduced and tumor appeared smoother after pCDNA3.1(+)miR132 transfection. Furthermore, overexpression of miR132 significantly suppressed the invasion of KYSE150 cells（55±1.6 vs 129.0±3.1, 124.0±2.8, P＜001), and miR132 decreased the expression of endogenous FOXA1 by targeting FOXA1 3'UTR. Conclusion: Esophageal carcinoma KYSE150 cells express low level of miR132, and overexpression of miR132 can negatively regulate the expression of FOXA1 and suppressed the invasion of KYSE150 cells.
2011, 18(5):490-495.
DOI: 10.3872/j.issn.1007-385X.2011.5.005
Abstract:
Objective:To study estrogen receptor (ER) subtype on the growth of breast cancer cell line MCF7 and the secretion of Th1 and Th2 cytokines in MCF7 tumor microenvironment. Methods: ERα or ERβ expression in MCF7 cells was silenced by RNA interference and MCF7 cells with different ERα/ERβ expression status were obtained. MTT test, flow cytometry and RTPCR assay were used to detect proliferation, cell cycle and expression of apoptosis suppressor genes. Secretion of IFNγ and IL4 in cell supernatant were analyzed by ELISA assay. Results: After RNA interference, protein levels of ERα or ERβ in MCF7 cells decreased by (77.7± 3.3 )% or (68.3±21)%, respectively. Compared to control group, after knocking down ERα gene expression, MCF7 cells grew slower (P＜0.05) and were arrested at phase G0～G1, expression of apoptosis suppressor gene XIAP decreased by (43.0±2.0)%. and the level of IFNγ increased by (1.89±0.34) times. However, after knocking down the ERβ gene expression, MCF7 grew faster (P<005), and the proportion of cells entering S phase increased, the expression of apoptosis suppressor genes Bcl2, Bclxl and XIAP increased by (1.28±0.21) times, (1.61±0.32) times and (1.65±0.29) times, respectively, while the level of IFNγ decreased by (28.0±4.0)%, compared to the control group. Conclusion: The expression status of ER subtype can affect the growth of MCF7 cells and induce the Th bias in microenvironment by regulating the autocrine level of IFNγ.
Objective:To study estrogen receptor (ER) subtype on the growth of breast cancer cell line MCF7 and the secretion of Th1 and Th2 cytokines in MCF7 tumor microenvironment. Methods: ERα or ERβ expression in MCF7 cells was silenced by RNA interference and MCF7 cells with different ERα/ERβ expression status were obtained. MTT test, flow cytometry and RTPCR assay were used to detect proliferation, cell cycle and expression of apoptosis suppressor genes. Secretion of IFNγ and IL4 in cell supernatant were analyzed by ELISA assay. Results: After RNA interference, protein levels of ERα or ERβ in MCF7 cells decreased by (77.7± 3.3 )% or (68.3±21)%, respectively. Compared to control group, after knocking down ERα gene expression, MCF7 cells grew slower (P＜0.05) and were arrested at phase G0～G1, expression of apoptosis suppressor gene XIAP decreased by (43.0±2.0)%. and the level of IFNγ increased by (1.89±0.34) times. However, after knocking down the ERβ gene expression, MCF7 grew faster (P<005), and the proportion of cells entering S phase increased, the expression of apoptosis suppressor genes Bcl2, Bclxl and XIAP increased by (1.28±0.21) times, (1.61±0.32) times and (1.65±0.29) times, respectively, while the level of IFNγ decreased by (28.0±4.0)%, compared to the control group. Conclusion: The expression status of ER subtype can affect the growth of MCF7 cells and induce the Th bias in microenvironment by regulating the autocrine level of IFNγ.
2011, 18(5):496-501.
DOI: 10.3872/j.issn.1007-385X.2011.5.006
Abstract:
Objective:To explore the effects of trichostatin A (TSA) and docetaxel (Doc) on apoptosis of lung adenocarcinoma A549 cells and the possible molecular mechanisms. Methods: A549 cells were treated with TSA alone or in combination with Doc, MTT assay was used to measure the proliferation of A549 cells; cell morphological changes were observed under light microscope; apoptosis was assessed using Hoechst 33258 staining and flow cytometry; cell cycle was analyzed by flow cytometry; the expression of acetylαtubulin and survivin protein and activation of caspase3 were detected by Western blotting. Results: 10 μg/ml Doc and 250 nmol/L TSA alone or in combination significantly inhibited the growth of A549 cells in a timedependent manner, and the combined treatment induced even higher inhibitory rate (\[656±31\]% vs \[30.6±2.1\]%, \[23.3±1.9\]%, P<0.05). In addition, TSA or Doc alone or in combination increased the apoptosis rate of A549 cells, and the combined treatment also induced higher apoptosis rate (\[58±3.6\]% vs \[17±2.2\]%, \[14±1.6\]%, P<0.05). The cell cycle was more markedly arrested in G2/M phase in combination treatment group (\[32.4±3.1\]% vs \[23.5±2.3\]%, \[10.5±1.5\]%, P<0.05), TSA combined with Doc increased the expression of acetylαtubulin, reduced the expression of survivin and promoted the activation of caspase3 (P<005). Conclusion: TSA combined with Doc can inhibit proliferation and induce apoptosis of A549 cells, which is related to the upregulation of acetylαtubulin, downregulation of survivin and increased activation of caspase3.
Objective:To explore the effects of trichostatin A (TSA) and docetaxel (Doc) on apoptosis of lung adenocarcinoma A549 cells and the possible molecular mechanisms. Methods: A549 cells were treated with TSA alone or in combination with Doc, MTT assay was used to measure the proliferation of A549 cells; cell morphological changes were observed under light microscope; apoptosis was assessed using Hoechst 33258 staining and flow cytometry; cell cycle was analyzed by flow cytometry; the expression of acetylαtubulin and survivin protein and activation of caspase3 were detected by Western blotting. Results: 10 μg/ml Doc and 250 nmol/L TSA alone or in combination significantly inhibited the growth of A549 cells in a timedependent manner, and the combined treatment induced even higher inhibitory rate (\[656±31\]% vs \[30.6±2.1\]%, \[23.3±1.9\]%, P<0.05). In addition, TSA or Doc alone or in combination increased the apoptosis rate of A549 cells, and the combined treatment also induced higher apoptosis rate (\[58±3.6\]% vs \[17±2.2\]%, \[14±1.6\]%, P<0.05). The cell cycle was more markedly arrested in G2/M phase in combination treatment group (\[32.4±3.1\]% vs \[23.5±2.3\]%, \[10.5±1.5\]%, P<0.05), TSA combined with Doc increased the expression of acetylαtubulin, reduced the expression of survivin and promoted the activation of caspase3 (P<005). Conclusion: TSA combined with Doc can inhibit proliferation and induce apoptosis of A549 cells, which is related to the upregulation of acetylαtubulin, downregulation of survivin and increased activation of caspase3.
2011, 18(5):502-507.
DOI: 10.3872/j.issn.1007-385X.2011.5.007
Abstract:
Objective:To investigate the effect of targeting silence of P65 gene by miRNA on adhesion and invasion of human triplenegative breast cancer (TNBC) cell line MDAMB231 in vitro and its possible mechanism. Methods: Three pairs of miRNAs specifically targeting P65 gene (P65miRNA1, P65miRNA2 and P65miRNA3) were designed and transfected into MDAMB231 cells, the level of P65 protein was detected by Western blotting, and P65miRNA with best silencing result was selected and used in the following experiments. The adhesive and invasive abilities of MDAMB231 cells before and after transfection were measured by adhesion assay and Transwell assay, respectively. mRNA expression of matrix metalloproteinase2 (MMP2) and MMP9 were detected by RTPCR and the activities of MMP2 and MMP9 were examined by gelatin zymography assay. Results: P65miRNA1, P65miRNA2 and P65miRNA3 plasmids were successfully constructed, P65miRNA1 and P65miRNA2, but not P65miRNA3, tranfection strongly inhibited the expression of P65 protein in MDAMB231 cells (>80%). The adhesion of MDAMB231 cells was not significantly influenced by P65miRNA1 and P65miRNA2 transfection (P>0.05), while the invasive ability of MDAMB231 cells was significantly reduced after P65miRNA1 or P65miRNA2 transfection (0.371±0.039, 0.309±0.046 vs 0.698±0065, P<0.05). Besides, mRNA expression of MMP2 (0.281±0.018, 0.478±0.023 vs 1.056±0.072, 1.128±0.059, P<0.05) and MMP9 (0.193±0.013, 0.371±0.035 vs 1.206±0.069, 1.089±0.057, P<0.05) and activities of MMP2 and MMP9 in MDAMB231 cells after P65miRNA1 or P65miRNA2 transfection were significantly decreased. Conclusion: Targeting silence of P65 expression can inhibit in vitro invasion of TNBC MDAMB231cells, which is related to the downregulation of MMP2 and MMP9 expression and their activities.
Objective:To investigate the effect of targeting silence of P65 gene by miRNA on adhesion and invasion of human triplenegative breast cancer (TNBC) cell line MDAMB231 in vitro and its possible mechanism. Methods: Three pairs of miRNAs specifically targeting P65 gene (P65miRNA1, P65miRNA2 and P65miRNA3) were designed and transfected into MDAMB231 cells, the level of P65 protein was detected by Western blotting, and P65miRNA with best silencing result was selected and used in the following experiments. The adhesive and invasive abilities of MDAMB231 cells before and after transfection were measured by adhesion assay and Transwell assay, respectively. mRNA expression of matrix metalloproteinase2 (MMP2) and MMP9 were detected by RTPCR and the activities of MMP2 and MMP9 were examined by gelatin zymography assay. Results: P65miRNA1, P65miRNA2 and P65miRNA3 plasmids were successfully constructed, P65miRNA1 and P65miRNA2, but not P65miRNA3, tranfection strongly inhibited the expression of P65 protein in MDAMB231 cells (>80%). The adhesion of MDAMB231 cells was not significantly influenced by P65miRNA1 and P65miRNA2 transfection (P>0.05), while the invasive ability of MDAMB231 cells was significantly reduced after P65miRNA1 or P65miRNA2 transfection (0.371±0.039, 0.309±0.046 vs 0.698±0065, P<0.05). Besides, mRNA expression of MMP2 (0.281±0.018, 0.478±0.023 vs 1.056±0.072, 1.128±0.059, P<0.05) and MMP9 (0.193±0.013, 0.371±0.035 vs 1.206±0.069, 1.089±0.057, P<0.05) and activities of MMP2 and MMP9 in MDAMB231 cells after P65miRNA1 or P65miRNA2 transfection were significantly decreased. Conclusion: Targeting silence of P65 expression can inhibit in vitro invasion of TNBC MDAMB231cells, which is related to the downregulation of MMP2 and MMP9 expression and their activities.
YAN Xiao-yan
,
CHENG Zhi-yong
,
LI Lin
,
LIANG Li-qing
,
WEI Zhan-hui
,
WANG Ya-Li
,
SI Miao
,
GAO Lian-bin
2011, 18(5):508-513.
DOI: 10.3872/j.issn.1007-385X.2011.5.008
Abstract:
Objective: To investigate the effect of phosphatase and tensin hemology deleted on chromosome ten gene (PTEN) on proliferation, apoptosis, VEGF (vascular endothelial growth factor) and its receptor VEGFR1 expression in human umbilical vein endothelial cell (HUVEC) line ECV304. Methods: Recombinant adenovirus containing green fluorescent protein (GFP) and PTEN (AdPTENGFP) or empty vector (AdGFP) were transfected into ECV304 cells; proliferation and apoptosis of ECV304 cells were measured by MTT assay, Hoechst3342 staining and flow cytometry, respectively; PTEN, VEGF, VEGFR1 mRNA expression levels in AdPTENGFPtransfected ECV304 cells were examined by quantitative PCR; and VEGF protein level in ECV304 cell supernatant was detected by ELISA. Chick chorioallantoic membrane (CAM) assay was used to study the effect of PTEN on angiogenesis. Results: AdPTENGFP transfection significantly inhibited the proliferation and induced apoptosis of ECV304 cells and the inhibitory rate and apoptotic rate were (50.38±5.42)% and (73.3±5.3)% at 5 d. VEGF and VEGFR1 mRNA expression levels were (13.40±1.32)% and (46.12±5.20)% of untransfected group after transfected with AdPTENGFP at MOI=100 in ECV304 cells. Furthermore, CAM assay results showed that AdPTENGFP transfection inhibited CAM angiogenesis in vivo. Conclusion: PTEN can inhibit the growth of and promote apoptosis of human umbilical vein endothelial ECV304 cells, which might be related to the downregulation of VEGF/VEGFR1 expression and the resulting angiogenesis inhibition.
Objective: To investigate the effect of phosphatase and tensin hemology deleted on chromosome ten gene (PTEN) on proliferation, apoptosis, VEGF (vascular endothelial growth factor) and its receptor VEGFR1 expression in human umbilical vein endothelial cell (HUVEC) line ECV304. Methods: Recombinant adenovirus containing green fluorescent protein (GFP) and PTEN (AdPTENGFP) or empty vector (AdGFP) were transfected into ECV304 cells; proliferation and apoptosis of ECV304 cells were measured by MTT assay, Hoechst3342 staining and flow cytometry, respectively; PTEN, VEGF, VEGFR1 mRNA expression levels in AdPTENGFPtransfected ECV304 cells were examined by quantitative PCR; and VEGF protein level in ECV304 cell supernatant was detected by ELISA. Chick chorioallantoic membrane (CAM) assay was used to study the effect of PTEN on angiogenesis. Results: AdPTENGFP transfection significantly inhibited the proliferation and induced apoptosis of ECV304 cells and the inhibitory rate and apoptotic rate were (50.38±5.42)% and (73.3±5.3)% at 5 d. VEGF and VEGFR1 mRNA expression levels were (13.40±1.32)% and (46.12±5.20)% of untransfected group after transfected with AdPTENGFP at MOI=100 in ECV304 cells. Furthermore, CAM assay results showed that AdPTENGFP transfection inhibited CAM angiogenesis in vivo. Conclusion: PTEN can inhibit the growth of and promote apoptosis of human umbilical vein endothelial ECV304 cells, which might be related to the downregulation of VEGF/VEGFR1 expression and the resulting angiogenesis inhibition.
2011, 18(5):514-518.
DOI: 10.3872/j.issn.1007-385X.2011.5.009
Abstract:
Objective:To observe the effect of celecoxib on apoptosis and autophagy of human gastric cancer cell line SGC7901, and to investigate the mechanism of apoptosis. Methods: SGC7901 cells were treated with different concentrations of celecoxib, proliferation of SGC7901 cells was studied by MTT assay, apoptosis was assessed by TUNEL, ultrastructure changes was observed by transmission electron microscopy, apoptotic rate was examined by flow cytometry, and expression of caspase8, caspase9 mRNA was analyzed by realtime quantitative PCR. Results: Celecoxib inhibited proliferation of SGC7901 cells in a time and dosedependent manner, with inhibitory rate of (85.6±451)% at 125 μmol/L celecoxib for 72 h. Celecoxib induced apoptosis of SGC7901 cells, typical apoptotic body and autophagosome were observed under TEM, and apoptotic rate increased from (2.2±1.32)% to (35.7±5.73)% (P<0.05) as detected by FCM. Expression of caspase8 and caspase9 mRNA increased sharply in SGC7901 cells treated with celecoxib in a time and dosedependent manner. Conclusion: Celecoxib can induce apoptosis of gastric cancer SGC7901 cells by activating caspase8 in the deathreceptor pathway and caspase9 in the mitochondrial pathway, and induce autophagic cell death.
Objective:To observe the effect of celecoxib on apoptosis and autophagy of human gastric cancer cell line SGC7901, and to investigate the mechanism of apoptosis. Methods: SGC7901 cells were treated with different concentrations of celecoxib, proliferation of SGC7901 cells was studied by MTT assay, apoptosis was assessed by TUNEL, ultrastructure changes was observed by transmission electron microscopy, apoptotic rate was examined by flow cytometry, and expression of caspase8, caspase9 mRNA was analyzed by realtime quantitative PCR. Results: Celecoxib inhibited proliferation of SGC7901 cells in a time and dosedependent manner, with inhibitory rate of (85.6±451)% at 125 μmol/L celecoxib for 72 h. Celecoxib induced apoptosis of SGC7901 cells, typical apoptotic body and autophagosome were observed under TEM, and apoptotic rate increased from (2.2±1.32)% to (35.7±5.73)% (P<0.05) as detected by FCM. Expression of caspase8 and caspase9 mRNA increased sharply in SGC7901 cells treated with celecoxib in a time and dosedependent manner. Conclusion: Celecoxib can induce apoptosis of gastric cancer SGC7901 cells by activating caspase8 in the deathreceptor pathway and caspase9 in the mitochondrial pathway, and induce autophagic cell death.
2011, 18(5):519-523.
DOI: 10.3872/j.issn.1007-385X.2011.5.010
Abstract:
Objective: To investigate the inhibitory effect of Brucea javanica oil emulsion on the growth of human largecell lung cancer NCIH460 cells in vitro and its mechanisms. Methods: NCIH460 cells were treated with different concentration of Brucea javanica oil emulsion (2.5, 5.0, 10, 20, 40 and 80 μg/ml) for 24, 48, and 72 h. The inhibitory effects of Brucea javanica oil emulsion on proliferation of NCIH460 cells was examined by MTT; morphological changes was observed by optical microscope after Giemsa staining; apoptotic rate of NCIH460 cells was examined by flow cytometry; and caspase3 activity in NCIH460 cells was detected with caspase3 activity kit. Results: Brucea javanica oil emulsion significantly inhibited the proliferation of NCIH460 in a dose and timedependent manner, and the inhibiting rate reached (91.07±1.60)% after 80 μg/ml Brucea javanica oil emulsion treatment for 72 h. Typical apoptotic morphological changes also appeared after Brucea javanica oil emulsion treatment. Flow cytometry results demonstrated that the apoptotic rates of NCIH460 cells were (45.23±4.30)%, (54.14±3.09)% and (61.57±7.28)% after 10, 20 and 40 μg/ml Brucea javanica oil emulsion treatment for 48 h, respectively (P＜0.01). In NCIH460 cells, the Brucea javanica oil emulsion increased the activity of caspase3 in a dosedependent manner. after 40 μg/ml Brucea javanica oil emulsion treatment, caspase 3 activity reached 1.07±0.07 μmol/μg, which was 4.97 fold higher than control group (P＜0.01). Conclusion:Brucea javanica oil emulsion can induce apoptosis and inhibit the proliferation of NCIH460 cells, possibly through increasing caspase3 activity.
Objective: To investigate the inhibitory effect of Brucea javanica oil emulsion on the growth of human largecell lung cancer NCIH460 cells in vitro and its mechanisms. Methods: NCIH460 cells were treated with different concentration of Brucea javanica oil emulsion (2.5, 5.0, 10, 20, 40 and 80 μg/ml) for 24, 48, and 72 h. The inhibitory effects of Brucea javanica oil emulsion on proliferation of NCIH460 cells was examined by MTT; morphological changes was observed by optical microscope after Giemsa staining; apoptotic rate of NCIH460 cells was examined by flow cytometry; and caspase3 activity in NCIH460 cells was detected with caspase3 activity kit. Results: Brucea javanica oil emulsion significantly inhibited the proliferation of NCIH460 in a dose and timedependent manner, and the inhibiting rate reached (91.07±1.60)% after 80 μg/ml Brucea javanica oil emulsion treatment for 72 h. Typical apoptotic morphological changes also appeared after Brucea javanica oil emulsion treatment. Flow cytometry results demonstrated that the apoptotic rates of NCIH460 cells were (45.23±4.30)%, (54.14±3.09)% and (61.57±7.28)% after 10, 20 and 40 μg/ml Brucea javanica oil emulsion treatment for 48 h, respectively (P＜0.01). In NCIH460 cells, the Brucea javanica oil emulsion increased the activity of caspase3 in a dosedependent manner. after 40 μg/ml Brucea javanica oil emulsion treatment, caspase 3 activity reached 1.07±0.07 μmol/μg, which was 4.97 fold higher than control group (P＜0.01). Conclusion:Brucea javanica oil emulsion can induce apoptosis and inhibit the proliferation of NCIH460 cells, possibly through increasing caspase3 activity.
2011, 18(5):524-527.
DOI: 10.3872/j.issn.1007-385X.2011.5.011
Abstract:
Objective:To investigate the expression, correlation and clinicopathological significance of enolaseα (ENO1) and tumor M2 pyruvate kinase (M2PK) in gastric cancer. Methods: Seventyeight paraffinembedded specimens of gastric tissues (55 gastric cancer and 23 gastric ulcer tissues) from the First Hospital of Lanzhou Universtiy (Jun. 2009 to Oct. 2010) were included in the present study. Expression of ENO1 and M2PK in gastric cancer and gastric ulcer tissues were detected by SP immunohistochemistry. Correlation between ENO1 expression and M2PK expression, and its implication in the clinicophathologic features of gastric cancer were analyzed. Results: The positive expression rate of ENO1 in gastric cancer tissues was significantly higher than in gastric ulcer tissues (67.3% vs 30.4%, P<0.01), and positively related to differentiation grade, depth of invasion, lymph node metastasis and TNM staging (all P<0.05). The positive expression rate of M2PK in gastric cancer tissues was significantly higher than in gastric ulcer (78.2% vs 391%, P<0.01), and positively related to differentiation grade (P<0.05), depth of invasion (all P<0.05). Expression of ENO1 was positively correlated to that of M2PK (r=0.5729, P<0.05). Conclusion: The upregulated expression of ENO1 and M2PK may participate in the oncogenesis and progression of gastric cancer. Combined detection of ENO1 and M2PK in gastric cancer tissues may be helpful in evaluating the prognosis of gastric cancer.
Objective:To investigate the expression, correlation and clinicopathological significance of enolaseα (ENO1) and tumor M2 pyruvate kinase (M2PK) in gastric cancer. Methods: Seventyeight paraffinembedded specimens of gastric tissues (55 gastric cancer and 23 gastric ulcer tissues) from the First Hospital of Lanzhou Universtiy (Jun. 2009 to Oct. 2010) were included in the present study. Expression of ENO1 and M2PK in gastric cancer and gastric ulcer tissues were detected by SP immunohistochemistry. Correlation between ENO1 expression and M2PK expression, and its implication in the clinicophathologic features of gastric cancer were analyzed. Results: The positive expression rate of ENO1 in gastric cancer tissues was significantly higher than in gastric ulcer tissues (67.3% vs 30.4%, P<0.01), and positively related to differentiation grade, depth of invasion, lymph node metastasis and TNM staging (all P<0.05). The positive expression rate of M2PK in gastric cancer tissues was significantly higher than in gastric ulcer (78.2% vs 391%, P<0.01), and positively related to differentiation grade (P<0.05), depth of invasion (all P<0.05). Expression of ENO1 was positively correlated to that of M2PK (r=0.5729, P<0.05). Conclusion: The upregulated expression of ENO1 and M2PK may participate in the oncogenesis and progression of gastric cancer. Combined detection of ENO1 and M2PK in gastric cancer tissues may be helpful in evaluating the prognosis of gastric cancer.
2011, 18(5):528-532.
DOI: 10.3872/j.issn.1007-385X.2011.5.012
Abstract:
Objective:To examine the expression of coinhibitory molecules B7H1, B7H4 and infiltration of T cell subsets in colorectal carcinoma tissues, and to explore their clinical significances. Methods: Fifty tumor tissue specimens and 5 paracancerous tissues of 50 colorectal carcinoma (CRC) patients (from Jan. 2003 to Dec. 2003) in Fourth Affiliated Hospital of Soochow University were collected in the study. Immunohistochemistry staining was used for the detection of B7H1, B7H4 expression and T cell subset infiltration in CRC tissues, correlation between B7H1, B7H4 expression and clinical parameters of CRC patients was further analyzed. Results: B7H1 and B7H4 expression was found strong in CRC tissues but negative in paracancerous tissues (P<0.05). B7H1 expression in colon cancer tissues was significantly higher than that in rectal cancer (P<0.05), B7H4 expression was positively correlated to the Duke’s stage (P<005). B7H1 but not B7H4 expression in CRC tissues was negatively correlated to the infiltration of CD3+T cell (P<0.05). B7H1 expression was reversely related to the patients’ prognosis (P<0.05), and patients with high levels of both B7H1 and B7H4 showed a lower overall survival rate than other patients (P<0.05). Conclusion: Coinhibitory molecules B7H1 and B7H4 were highly expressed in human CRC tissues, and correlated to patients’ overall survival, and the joint detection of these molecules may have clinical value for diagnosis and prognosis of colorectal carcinoma.
Objective:To examine the expression of coinhibitory molecules B7H1, B7H4 and infiltration of T cell subsets in colorectal carcinoma tissues, and to explore their clinical significances. Methods: Fifty tumor tissue specimens and 5 paracancerous tissues of 50 colorectal carcinoma (CRC) patients (from Jan. 2003 to Dec. 2003) in Fourth Affiliated Hospital of Soochow University were collected in the study. Immunohistochemistry staining was used for the detection of B7H1, B7H4 expression and T cell subset infiltration in CRC tissues, correlation between B7H1, B7H4 expression and clinical parameters of CRC patients was further analyzed. Results: B7H1 and B7H4 expression was found strong in CRC tissues but negative in paracancerous tissues (P<0.05). B7H1 expression in colon cancer tissues was significantly higher than that in rectal cancer (P<0.05), B7H4 expression was positively correlated to the Duke’s stage (P<005). B7H1 but not B7H4 expression in CRC tissues was negatively correlated to the infiltration of CD3+T cell (P<0.05). B7H1 expression was reversely related to the patients’ prognosis (P<0.05), and patients with high levels of both B7H1 and B7H4 showed a lower overall survival rate than other patients (P<0.05). Conclusion: Coinhibitory molecules B7H1 and B7H4 were highly expressed in human CRC tissues, and correlated to patients’ overall survival, and the joint detection of these molecules may have clinical value for diagnosis and prognosis of colorectal carcinoma.
2011, 18(5):533-538.
DOI: 10.3872/j.issn.1007-385X.2011.5.013
Abstract:
Objective:To investigate the infiltration of Foxp3+ cells and expression of Foxp3 protein in breast carcinoma tissues and their clinical significance. Methods: One hundred and sixty breast carcinoma and 22 paracancerous tissues (who had been diagnosed in Shanghai Huangpu Center Hospital from Jan. 2005 to Dec. 2005) were included in present study. The infiltration of Foxp3+ cells and expression of Foxp3 protein in breast carcinoma and paracancerous tissues were detected by immunohistochemistry, and their relationships to clinicopathologic features and expression of ER,PR,P53,Bcl2,HER2 in breast carcinoma were analyzed. Results: Both positive rate of Foxp3+ cell infiltration in mesenchyma tissues and Foxp3 protein in parenchyma tissues were significantly higher than those in normal paracancerous tissues (P<0.05), but there was no correlation between the Foxp3+ cell infiltration and Foxp3 protein expression in breast carcinoma tissues (P>0.05). Foxp3+ cell infiltration in mesenchyma tissues was positively correlated to lymph node metastasis, histological grade and P53 overexpression (P<0.05); Foxp3 protein expression in parenchyma tissues was positively correlated to lymph node metastasis, histological grade, pTNM stage and P53 overexpression (P<005), but not to overexpression of ER, PR, HER2 and Bcl2 (P>0.05). Univariate survival analysis indicated that Foxp3 expression in breast carcinoma was associated with 5year overall survival rate, while multivariate analysis indicated that Foxp3 expression was not an independent prognostic factor for 5year overall survival rate in breast carcinoma (P>005). Conclusion: Foxp3 protein is overexpressed in breast carcinoma tissues, which might be a potential biomarker but not an independent prognostic factor in breast carcinoma.
Objective:To investigate the infiltration of Foxp3+ cells and expression of Foxp3 protein in breast carcinoma tissues and their clinical significance. Methods: One hundred and sixty breast carcinoma and 22 paracancerous tissues (who had been diagnosed in Shanghai Huangpu Center Hospital from Jan. 2005 to Dec. 2005) were included in present study. The infiltration of Foxp3+ cells and expression of Foxp3 protein in breast carcinoma and paracancerous tissues were detected by immunohistochemistry, and their relationships to clinicopathologic features and expression of ER,PR,P53,Bcl2,HER2 in breast carcinoma were analyzed. Results: Both positive rate of Foxp3+ cell infiltration in mesenchyma tissues and Foxp3 protein in parenchyma tissues were significantly higher than those in normal paracancerous tissues (P<0.05), but there was no correlation between the Foxp3+ cell infiltration and Foxp3 protein expression in breast carcinoma tissues (P>0.05). Foxp3+ cell infiltration in mesenchyma tissues was positively correlated to lymph node metastasis, histological grade and P53 overexpression (P<0.05); Foxp3 protein expression in parenchyma tissues was positively correlated to lymph node metastasis, histological grade, pTNM stage and P53 overexpression (P<005), but not to overexpression of ER, PR, HER2 and Bcl2 (P>0.05). Univariate survival analysis indicated that Foxp3 expression in breast carcinoma was associated with 5year overall survival rate, while multivariate analysis indicated that Foxp3 expression was not an independent prognostic factor for 5year overall survival rate in breast carcinoma (P>005). Conclusion: Foxp3 protein is overexpressed in breast carcinoma tissues, which might be a potential biomarker but not an independent prognostic factor in breast carcinoma.
14 Expression of Hedgehog signaling pathway members in colon cancer and their clinical significances
2011, 18(5):539-543.
DOI: 10.3872/j.issn.1007-385X.2011.5.014
Abstract:
Objective:To study the aberrant activation of Hedgehog signaling pathway in colon cancer and its clinical significances. Methods:Sixtysix colon cancer and 20 paracancerous tissue samples (Mar. 2009 to Jun. 2010, Xinhua Hospital, Shanghai Jiaotong University School of Medicine) were included in the study. Expression of the Hedgehog signaling pathway members, SHH, PTCH1, Gli1 and SuFu, in colon cancer tissues were detected by immunohistochemistry, and their relationship with clinicopathologic characteristics of colon cancer was examined. Results: In colon cancer tissues, SHH and PTCH1 were highly expressed, while PTCH1 and SuFu were weakly expressed, with their expression rates being 64%, 36%, 72%, and 20%, respectively. In paracancerous tissues SHH and PTCH1 were weakly expressed, and Gli1 and SuFu were not expressed. PTCH1 and Gli1 exression were related to infiltration depth of colon cancer (P=0023, P=0.040), and showed no relationship with age, gender, pathology. SHH and SuFu showed no relationship with age, gender, pathology, infiltration depth, etc.Conclusion: Hedgehog pathway members are highly expressed in colon cancer, which may be involved in oncogenesis and development of colon cancer.
Objective:To study the aberrant activation of Hedgehog signaling pathway in colon cancer and its clinical significances. Methods:Sixtysix colon cancer and 20 paracancerous tissue samples (Mar. 2009 to Jun. 2010, Xinhua Hospital, Shanghai Jiaotong University School of Medicine) were included in the study. Expression of the Hedgehog signaling pathway members, SHH, PTCH1, Gli1 and SuFu, in colon cancer tissues were detected by immunohistochemistry, and their relationship with clinicopathologic characteristics of colon cancer was examined. Results: In colon cancer tissues, SHH and PTCH1 were highly expressed, while PTCH1 and SuFu were weakly expressed, with their expression rates being 64%, 36%, 72%, and 20%, respectively. In paracancerous tissues SHH and PTCH1 were weakly expressed, and Gli1 and SuFu were not expressed. PTCH1 and Gli1 exression were related to infiltration depth of colon cancer (P=0023, P=0.040), and showed no relationship with age, gender, pathology. SHH and SuFu showed no relationship with age, gender, pathology, infiltration depth, etc.Conclusion: Hedgehog pathway members are highly expressed in colon cancer, which may be involved in oncogenesis and development of colon cancer.
2011, 18(5):544-547.
DOI: 10.3872/j.issn.1007-385X.2011.5.015
Abstract:
Objective:To evaluate the propotion of CD4+CD25+CD127low/- regulatory T cells in peripheral blood of papillary thyroid carcinoma (PTC) patients and its clinical significance. Methods: Forty patients with PTC and 30 patients with thyroid adenoma were included in this study. They were all treated in Renji Hospital, Medical School of Shanghai Jiaotong University between August 2010 to February 2011. The proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC and thyroid adenoma patients was evaluated by flow cytometry, and their relationship to clinicopathological features of PTC was studied. Results: Compared to thyroid adenoma patients, the proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC patients was significantly increased (\[7.836±1.668\]% vs \[5.365±1.156\]%, P<005). The proportion of CD4+CD25+CD127low/-Treg was related to clinical tumor stage and cervical lymph node metastasis of PTC patients (all P<0.05), but not related to age, gender, and tumor size (P>0.05). Conclusion: The proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC patients was significantly increased, and related to clinical tumor stage and cervical lymph node metastasis of PTC patients.
Objective:To evaluate the propotion of CD4+CD25+CD127low/- regulatory T cells in peripheral blood of papillary thyroid carcinoma (PTC) patients and its clinical significance. Methods: Forty patients with PTC and 30 patients with thyroid adenoma were included in this study. They were all treated in Renji Hospital, Medical School of Shanghai Jiaotong University between August 2010 to February 2011. The proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC and thyroid adenoma patients was evaluated by flow cytometry, and their relationship to clinicopathological features of PTC was studied. Results: Compared to thyroid adenoma patients, the proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC patients was significantly increased (\[7.836±1.668\]% vs \[5.365±1.156\]%, P<005). The proportion of CD4+CD25+CD127low/-Treg was related to clinical tumor stage and cervical lymph node metastasis of PTC patients (all P<0.05), but not related to age, gender, and tumor size (P>0.05). Conclusion: The proportion of CD4+CD25+CD127low/-Treg in peripheral blood of PTC patients was significantly increased, and related to clinical tumor stage and cervical lymph node metastasis of PTC patients.
2011, 18(5):548-551.
DOI: 10.3872/j.issn.1007-385X.2011.5.016
Abstract:
Objective:To investigate the clinical significance of circulating endothelial progenitor cells (EPCs) counting in diagnosis and prognosis of hepatocellular carcinoma (HCC). Methods: Tirtynine patients with HCC and 20 healthy individuals were recruited from Shanghai First People’s Hospital Baoshan Branch (Jan. 2008 to Dec. 2010) in this study. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation, then stained with antibodies against CD34, CD133, VEGFR2 which were conjugated to FITC, PE and APC respectively. Circulating EPCs were characterized as CD34+CD133+VEGFR2+ cells and its proportion in peripheral blood mononuclear cells was detected by flow cytometry. Relationship between the proportion of CD34+CD133+VEGFR2+ EPC and clinical stage, etc. of HCC was analyzed. Results:The level of circulating CD34+CD133+VEGFR2+ EPCs in HCC patients was significantly higher than that in healthy control(0.138±0.05)%,vs (0.027±0.013)% (P<0.05). Furthermore, the level of circulating EPCs in HCC patients was positively correlated to clinical stages. Conclusion: Circulating CD34+CD133+VEGFR2+ EPCs proportion is upregulated in HCC patients, and related to clinical stage of HCC, which may be significant in diagnosis and prognosis of HCC.
Objective:To investigate the clinical significance of circulating endothelial progenitor cells (EPCs) counting in diagnosis and prognosis of hepatocellular carcinoma (HCC). Methods: Tirtynine patients with HCC and 20 healthy individuals were recruited from Shanghai First People’s Hospital Baoshan Branch (Jan. 2008 to Dec. 2010) in this study. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation, then stained with antibodies against CD34, CD133, VEGFR2 which were conjugated to FITC, PE and APC respectively. Circulating EPCs were characterized as CD34+CD133+VEGFR2+ cells and its proportion in peripheral blood mononuclear cells was detected by flow cytometry. Relationship between the proportion of CD34+CD133+VEGFR2+ EPC and clinical stage, etc. of HCC was analyzed. Results:The level of circulating CD34+CD133+VEGFR2+ EPCs in HCC patients was significantly higher than that in healthy control(0.138±0.05)%,vs (0.027±0.013)% (P<0.05). Furthermore, the level of circulating EPCs in HCC patients was positively correlated to clinical stages. Conclusion: Circulating CD34+CD133+VEGFR2+ EPCs proportion is upregulated in HCC patients, and related to clinical stage of HCC, which may be significant in diagnosis and prognosis of HCC.
2011, 18(5):552-554.
DOI: 10.3872/j.issn.1007-385X.2011.5.017
Abstract:
目的：研究食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）及癌旁黏膜组织中的细胞外信号调节激酶（extracellular signal ragulated kinase，ERK）mRNA的表达，探讨ERK在ESCC发生、发展中的作用及其意义。方法：收集河北医科大学第四医院胸外科40例ESCC患者癌组织标本及25例患者的癌旁组织标本，RTPCR检测ESCC与癌旁组织中ERK mRNA的表达情况，分析其与ESCC临床病理特征的关系。结果：ESCC组织中ERK mRNA的表达明显增高，而在癌旁组织中呈现低表达（0.656±0.055 vs 0.450±0.070，P<0.01）。ESCC组织中ERK mRNA的表达与ESCC患者的年龄、性别和组织分化程度无关（P>0.05），而与临床分期、淋巴结转移显著相关（P<0.05）。结论：ESCC组织高表达ERK mRNA，ERK可能在ESCC发生、发展中起重要的作用。
目的：研究食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）及癌旁黏膜组织中的细胞外信号调节激酶（extracellular signal ragulated kinase，ERK）mRNA的表达，探讨ERK在ESCC发生、发展中的作用及其意义。方法：收集河北医科大学第四医院胸外科40例ESCC患者癌组织标本及25例患者的癌旁组织标本，RTPCR检测ESCC与癌旁组织中ERK mRNA的表达情况，分析其与ESCC临床病理特征的关系。结果：ESCC组织中ERK mRNA的表达明显增高，而在癌旁组织中呈现低表达（0.656±0.055 vs 0.450±0.070，P<0.01）。ESCC组织中ERK mRNA的表达与ESCC患者的年龄、性别和组织分化程度无关（P>0.05），而与临床分期、淋巴结转移显著相关（P<0.05）。结论：ESCC组织高表达ERK mRNA，ERK可能在ESCC发生、发展中起重要的作用。
2011, 18(5):555-560.
DOI: 10.3872/j.issn.1007-385X.2011.5.018
Abstract:
条件复制型腺病毒（conditionally replicating adenovirus，CRAd）基因治疗肿瘤的研究主要集中于提高其靶向性、增强肿瘤杀伤特异性，以及降低毒性作用三个方面。在提高CRAd靶向性的研究中，早期策略主要集中在腺病毒早期复制必需基因 E1a/E1b 突变或缺失的调控以及肿瘤或组织特异性启动子对其转录的调控；近年来利用组织特异性microRNA对病毒早期复制必需基因的转录后调控以及病毒外壳蛋白修饰的转导调控已逐渐成为研究的热点。在提高CRAd的肿瘤杀伤方面，除删除病毒自身凋亡抑制基因与导入外源性治疗基因两种方法以外，改造外壳纤毛提高病毒对靶细胞的亲嗜性及构建靶向肿瘤干细胞的CRAd成为新的关注点。同时，随着对腺病毒结构认识的逐步加深，修饰其外壳六邻体蛋白及其他外壳蛋白以降低CRAd的肝嗜性与免疫原性也正成为降低CRAd毒性作用的研究热点。
条件复制型腺病毒（conditionally replicating adenovirus，CRAd）基因治疗肿瘤的研究主要集中于提高其靶向性、增强肿瘤杀伤特异性，以及降低毒性作用三个方面。在提高CRAd靶向性的研究中，早期策略主要集中在腺病毒早期复制必需基因 E1a/E1b 突变或缺失的调控以及肿瘤或组织特异性启动子对其转录的调控；近年来利用组织特异性microRNA对病毒早期复制必需基因的转录后调控以及病毒外壳蛋白修饰的转导调控已逐渐成为研究的热点。在提高CRAd的肿瘤杀伤方面，除删除病毒自身凋亡抑制基因与导入外源性治疗基因两种方法以外，改造外壳纤毛提高病毒对靶细胞的亲嗜性及构建靶向肿瘤干细胞的CRAd成为新的关注点。同时，随着对腺病毒结构认识的逐步加深，修饰其外壳六邻体蛋白及其他外壳蛋白以降低CRAd的肝嗜性与免疫原性也正成为降低CRAd毒性作用的研究热点。
2011, 18(5):561-564.
DOI: 10.3872/j.issn.1007-385X.2011.5.019
Abstract:
指数富集的配基系统进化技术（systematic evolution of ligands by exponential enrichment，SELEX）应用人工合成的随机寡核苷酸文库，通过筛选、分离、富集获得能与各种配体特异结合的寡核苷酸适配子(aptamer)。适配子能特异性结合多肽、蛋白质等多种靶分子，且具有高亲和力。细胞SELEX技术在SELEX基础上发展起来，它以活细胞为基础，可以在对肿瘤细胞分子标志物一无所知的情况下有效地筛选出肿瘤细胞特异性适配子，是一个潜在的发现肿瘤细胞新标志物的理想策略。肿瘤细胞特异性适配子作为分子探针，在肿瘤基础研究、早期诊断，以及靶向治疗方面展示了巨大的应用前景。目前已筛选出淋巴细胞白血病、肝癌等多种肿瘤细胞的适配子，这些适配子已应用在肿瘤细胞检测，肿瘤早期发现、诊断、游离肿瘤细胞的分选和富集，以及肿瘤细胞靶向药物递送等方面，并且由于适配子适用范围广泛、易于修饰等特点，显示出良好的敏感性、特异性和其他独特的优越性。适配子在肿瘤研究中受到越来越多的关注，在肿瘤检测、诊断和治疗等方面发挥重要作用。
指数富集的配基系统进化技术（systematic evolution of ligands by exponential enrichment，SELEX）应用人工合成的随机寡核苷酸文库，通过筛选、分离、富集获得能与各种配体特异结合的寡核苷酸适配子(aptamer)。适配子能特异性结合多肽、蛋白质等多种靶分子，且具有高亲和力。细胞SELEX技术在SELEX基础上发展起来，它以活细胞为基础，可以在对肿瘤细胞分子标志物一无所知的情况下有效地筛选出肿瘤细胞特异性适配子，是一个潜在的发现肿瘤细胞新标志物的理想策略。肿瘤细胞特异性适配子作为分子探针，在肿瘤基础研究、早期诊断，以及靶向治疗方面展示了巨大的应用前景。目前已筛选出淋巴细胞白血病、肝癌等多种肿瘤细胞的适配子，这些适配子已应用在肿瘤细胞检测，肿瘤早期发现、诊断、游离肿瘤细胞的分选和富集，以及肿瘤细胞靶向药物递送等方面，并且由于适配子适用范围广泛、易于修饰等特点，显示出良好的敏感性、特异性和其他独特的优越性。适配子在肿瘤研究中受到越来越多的关注，在肿瘤检测、诊断和治疗等方面发挥重要作用。
2011, 18(5):565-568.
DOI: 10.3872/j.issn.1007-385X.2011.5.020
Abstract:
热激蛋白（heat shock proteins, HSP，曾称热休克蛋白）是一类广泛存在于原核、真核生物细胞内的热应激蛋白质，和HSP受体结合发挥应激、抗感染、抗肿瘤作用。与肿瘤免疫相关的HSP受体主要包括TLR2/4、CD91、SR、CCR5等。TLR2/4一方面可与CD14 形成受体复合物结合HSP，促进树突状细胞（DC）和免疫效应细胞聚集，激活抗肿瘤免疫应答；另一方面也可在肿瘤细胞表面高表达，促进肿瘤生长分化。CD91能够发挥肿瘤抗原提呈作用，作为信号性受体启动抗肿瘤免疫应答；但表达于肿瘤细胞上的CD91往往会促进肿瘤的迁移和侵袭。SR参与内源性抗原识别和交叉提呈，可以识别肿瘤细胞，进而激活大量CD8+T细胞发挥抗肿瘤作用。CCR5参与致癌机制，是多种肿瘤发生、发展和预后判断的新型生物学标志。随着研究的深入HSP受体有望成为未来肿瘤防治的新突破口。
热激蛋白（heat shock proteins, HSP，曾称热休克蛋白）是一类广泛存在于原核、真核生物细胞内的热应激蛋白质，和HSP受体结合发挥应激、抗感染、抗肿瘤作用。与肿瘤免疫相关的HSP受体主要包括TLR2/4、CD91、SR、CCR5等。TLR2/4一方面可与CD14 形成受体复合物结合HSP，促进树突状细胞（DC）和免疫效应细胞聚集，激活抗肿瘤免疫应答；另一方面也可在肿瘤细胞表面高表达，促进肿瘤生长分化。CD91能够发挥肿瘤抗原提呈作用，作为信号性受体启动抗肿瘤免疫应答；但表达于肿瘤细胞上的CD91往往会促进肿瘤的迁移和侵袭。SR参与内源性抗原识别和交叉提呈，可以识别肿瘤细胞，进而激活大量CD8+T细胞发挥抗肿瘤作用。CCR5参与致癌机制，是多种肿瘤发生、发展和预后判断的新型生物学标志。随着研究的深入HSP受体有望成为未来肿瘤防治的新突破口。
2011, 18(5):569-573.
DOI: 10.3872/j.issn.1007-385X.2011.5.021
Abstract:
肿瘤转移与肿瘤微环境中成纤维细胞、转化生长因子、肿瘤相关巨噬细胞、趋化因子及其受体、凝血酶等多种因素密切相关。成纤维细胞通过促进肿瘤血管生成、促进癌细胞与细胞外基质黏附、促进细胞外基质降解等环节参与肿瘤的转移。TGFβ是由巨噬细胞、间质细胞和肿瘤细胞产生，它能对抗血管内皮的紧密连接和黏附连接，使毛细血管壁完整性受到破坏，从而导致毛细血管通透性增加，使肿瘤细胞从血管中游出进入器官组织中形成种植转移。肿瘤相关性巨噬细胞可合成和分泌EGF等细胞因子，引导肿瘤细胞穿越血管壁，促进肿瘤的转移。趋化因子及其受体对肿瘤细胞的迁移起着决定性的作用。凝血酶能通过影响微环境中其他细胞的行为而为肿瘤转移提供一个相容的环境。明晰肿瘤转移与肿瘤微环境的关系，进而明确在肿瘤发生、发展、转移过程中发挥重要作用的关键分子，寻找其相对应的靶点，对于肿瘤的诊断及治疗具有重要的作用。
肿瘤转移与肿瘤微环境中成纤维细胞、转化生长因子、肿瘤相关巨噬细胞、趋化因子及其受体、凝血酶等多种因素密切相关。成纤维细胞通过促进肿瘤血管生成、促进癌细胞与细胞外基质黏附、促进细胞外基质降解等环节参与肿瘤的转移。TGFβ是由巨噬细胞、间质细胞和肿瘤细胞产生，它能对抗血管内皮的紧密连接和黏附连接，使毛细血管壁完整性受到破坏，从而导致毛细血管通透性增加，使肿瘤细胞从血管中游出进入器官组织中形成种植转移。肿瘤相关性巨噬细胞可合成和分泌EGF等细胞因子，引导肿瘤细胞穿越血管壁，促进肿瘤的转移。趋化因子及其受体对肿瘤细胞的迁移起着决定性的作用。凝血酶能通过影响微环境中其他细胞的行为而为肿瘤转移提供一个相容的环境。明晰肿瘤转移与肿瘤微环境的关系，进而明确在肿瘤发生、发展、转移过程中发挥重要作用的关键分子，寻找其相对应的靶点，对于肿瘤的诊断及治疗具有重要的作用。
2011, 18(5):574-580.
DOI: 10.3872/j.issn.1007-385X.2011.5.022
Abstract:
p57Kip2属于细胞周期依赖激酶抑制因子（cyclin dependent kinases inhibitor，CDKI）Cip/Kip家族的成员之一，CKI能竞争性地与细胞周期蛋白依赖性激酶（cyclin dependent kinase，CDK）结合抑制其活性，从而调整细胞周期。与家族其它成员相比，p57Kip2在结构和功能有其特殊性。p57Kip2通过多种机制参与肿瘤的发生、侵袭和转移过程，Cip/Kip蛋白能在不同水平抑制Rho/ROCK/LIM/coffilin信号通路而参与肿瘤的形成、侵袭和转移，但在不同的细胞定位和调控下，p57Kip2在肿瘤的侵袭和转移中扮演双重角色。p57Kip2在细胞分化、凋亡上也具有重要作用，p57Kip2的表达异常使细胞不分化和过度增殖而形成肿瘤，p57Kip2在细胞凋亡中的作用也许可以作为肿瘤治疗的靶点。多功能的p57Kip2通过印记丢失、杂合性缺失、启动子甲基化、组蛋白去乙酰化和microRNA的调控等参与肿瘤形成的多个过程。本文就p57Kip2在以上几个方面的研究进展做一综述。
p57Kip2属于细胞周期依赖激酶抑制因子（cyclin dependent kinases inhibitor，CDKI）Cip/Kip家族的成员之一，CKI能竞争性地与细胞周期蛋白依赖性激酶（cyclin dependent kinase，CDK）结合抑制其活性，从而调整细胞周期。与家族其它成员相比，p57Kip2在结构和功能有其特殊性。p57Kip2通过多种机制参与肿瘤的发生、侵袭和转移过程，Cip/Kip蛋白能在不同水平抑制Rho/ROCK/LIM/coffilin信号通路而参与肿瘤的形成、侵袭和转移，但在不同的细胞定位和调控下，p57Kip2在肿瘤的侵袭和转移中扮演双重角色。p57Kip2在细胞分化、凋亡上也具有重要作用，p57Kip2的表达异常使细胞不分化和过度增殖而形成肿瘤，p57Kip2在细胞凋亡中的作用也许可以作为肿瘤治疗的靶点。多功能的p57Kip2通过印记丢失、杂合性缺失、启动子甲基化、组蛋白去乙酰化和microRNA的调控等参与肿瘤形成的多个过程。本文就p57Kip2在以上几个方面的研究进展做一综述。
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.