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Volume 25,Issue 4,2018 Table of Contents
2018, 25(4):321-328.
DOI: 10.3872/j.issn.1007-385X.2018.04.001
Abstract:
[Abstract] Glioblastoma multiforme （GBM）is a malignant tumor of central nervous system with high incidence, aggressive and poor prognosis. Since temozolomide was approved by FDA in 2005, there is no new curative strategy with obvious improvement in therapeu- tic effect. With the developments in molecular biology, tumor immunology and immunotherapy technology, the discovery of new molec- ular targets, breakthrough in central immunization exemption theory, and advance in gene transduction and cell technology, GBM im- munotherapy ushered in a new breakthrough in immunotherapy. Cellular immunotherapy, presented by chimeric antigen receptor-modi- fied T cell (CAR-T) therapy, has exhibiteditsprominentapplicationprospectinGBMinvitro experimentstargetingEGFRvIII, IL-13Rα2, HER2, erythropoietin-producing hepatocellular carcinoma A2 (EphA2), animal models and early clinical trials. However, the GBM mo- lecular heterogeneity, immunosuppressive microenvironment and blood-brain barrier have presented challenge for CAR-T going into the first-line clinical treatment. Researchers are now exploring key antigens of oncogenic phenotype, designing optimal combination of target antigens to prevent the immune escape, improving CAR-T passing through blood-brain barrier and invading tumor tissue, finding the best route for cell deliver, and optimizing evaluation system for central nervous system (CNS) immunotherapy. It is believed that the breakthrough of CAR-T cell immunotherapy will finally help GBM patients pursuing a beautiful life.
[Abstract] Glioblastoma multiforme （GBM）is a malignant tumor of central nervous system with high incidence, aggressive and poor prognosis. Since temozolomide was approved by FDA in 2005, there is no new curative strategy with obvious improvement in therapeu- tic effect. With the developments in molecular biology, tumor immunology and immunotherapy technology, the discovery of new molec- ular targets, breakthrough in central immunization exemption theory, and advance in gene transduction and cell technology, GBM im- munotherapy ushered in a new breakthrough in immunotherapy. Cellular immunotherapy, presented by chimeric antigen receptor-modi- fied T cell (CAR-T) therapy, has exhibiteditsprominentapplicationprospectinGBMinvitro experimentstargetingEGFRvIII, IL-13Rα2, HER2, erythropoietin-producing hepatocellular carcinoma A2 (EphA2), animal models and early clinical trials. However, the GBM mo- lecular heterogeneity, immunosuppressive microenvironment and blood-brain barrier have presented challenge for CAR-T going into the first-line clinical treatment. Researchers are now exploring key antigens of oncogenic phenotype, designing optimal combination of target antigens to prevent the immune escape, improving CAR-T passing through blood-brain barrier and invading tumor tissue, finding the best route for cell deliver, and optimizing evaluation system for central nervous system (CNS) immunotherapy. It is believed that the breakthrough of CAR-T cell immunotherapy will finally help GBM patients pursuing a beautiful life.
2018, 25(4):329-333.
DOI: 10.3872/j.issn.1007-385X.2018.04.002
Abstract:
[Abstract] Glioblastoma multiforme (GBM) is the most common malignant tumor of the central nervous system. Despite advances in traditional treatment strategies that combine surgery with radiotherapy and chemotherapy, GBM remains one of the most lethal diseases with dismal prognosis. Epithelial-interstitial transformation (EMT) is an important biological process for the invasion and migration of malignant tumors derived from epithelial cells, which is closely related to the pathological behaviors of GBM including invasion, migra- tion, resistance to chemotherapy and radiotherapy. This review will introduce the concept of EMT and its pathophysiological process, especially the latest findings related to the GBM biology. Besides, gene regulation and signaling pathways of EMT (such as matrix me- talloproteinases [MMP], TGF-β, transcription factors Snail and Twist etc.) participated in GBM are also introduced, thereby providing a new insight into the fundamental researches and clinical treatments of GBM.
[Abstract] Glioblastoma multiforme (GBM) is the most common malignant tumor of the central nervous system. Despite advances in traditional treatment strategies that combine surgery with radiotherapy and chemotherapy, GBM remains one of the most lethal diseases with dismal prognosis. Epithelial-interstitial transformation (EMT) is an important biological process for the invasion and migration of malignant tumors derived from epithelial cells, which is closely related to the pathological behaviors of GBM including invasion, migra- tion, resistance to chemotherapy and radiotherapy. This review will introduce the concept of EMT and its pathophysiological process, especially the latest findings related to the GBM biology. Besides, gene regulation and signaling pathways of EMT (such as matrix me- talloproteinases [MMP], TGF-β, transcription factors Snail and Twist etc.) participated in GBM are also introduced, thereby providing a new insight into the fundamental researches and clinical treatments of GBM.
2018, 25(4):334-339.
DOI: 10.3872/j.issn.1007-385X.2018.04.003
Abstract:
Objective:To prepare the third generation CAR-T cells targeting EGFRvⅢ (EGFRvⅢCAR-T) and to detect its specific killing effect against EGFRvⅢ + U87 cells in vitro and in vivo. Methods: Human CD3 + T cells were transfected with lentiviral EGFRv Ⅲ/3CAR, which was generated by calcium phosphate co-precipitation of three plasmids. The expression of EGFRvⅢ/3CAR in T cells was detected by Western blotting and flow cytometry. In vitro killing effect of EGFRvⅢ/3CAR-T cells on EGFRvⅢ + U87 cells was de- tected by 51 Cr release assay. The secretion of cytokine IFN-γ of EGFRvⅢ/3CAR-T cells was detected by ELISA. Nude mouse xeno- graft model was constructed to detect the in vivo cytotoxicity of EGFRvⅢ/3CAR-T cells on xenograft tumor. Results: The EGFRvⅢ/ 3CAR lentivirus was successfully packaged with an average titer of 5×10 6 TU/ml. Western blotting showed that a protein band of ap- proximate 58 000 molecular weight was observed in EGFRvⅢ/3CAR-T cells but absent in untransfected T cells. Flow cytometry indi- cated the average transduction efficiency of EGFRvⅢ/3CAR was 52.3%. 51 Cr release assay showed that the specific killing effect of EGFRvⅢ/3CAR-T cells was positively correlated with E/T ratio (E∶T=4∶1, 8∶1, 16∶1, 32∶1). ELISA showed that cytokine IFN-γ se- cretion was (1 836±148.2) pg/ml, which was significantly different from that of NT T and GFP + T cells (P<0.01). The specific killing ac- tivity of EGFRvⅢ/3CAR-T cells and IFN-γ secretion were both dependent on the expression level of EGFRvⅢ in U87 cells. The tu-mor growth monitoring results showed that the tumor volume of EGFRvⅢ/3CAR-T cell group was significantly different from that of GFP + T cell group and PBS group around 3 weeks after injection (P<0.01). Conclusion: EGFRvⅢ/3CAR-T cells demonstrated specific antitumor effectagainstEGFRvⅢ + U87cellsbothinvitro and in vivo, providing basis for immunotherapyofgliomainfuture clinical use.
Objective:To prepare the third generation CAR-T cells targeting EGFRvⅢ (EGFRvⅢCAR-T) and to detect its specific killing effect against EGFRvⅢ + U87 cells in vitro and in vivo. Methods: Human CD3 + T cells were transfected with lentiviral EGFRv Ⅲ/3CAR, which was generated by calcium phosphate co-precipitation of three plasmids. The expression of EGFRvⅢ/3CAR in T cells was detected by Western blotting and flow cytometry. In vitro killing effect of EGFRvⅢ/3CAR-T cells on EGFRvⅢ + U87 cells was de- tected by 51 Cr release assay. The secretion of cytokine IFN-γ of EGFRvⅢ/3CAR-T cells was detected by ELISA. Nude mouse xeno- graft model was constructed to detect the in vivo cytotoxicity of EGFRvⅢ/3CAR-T cells on xenograft tumor. Results: The EGFRvⅢ/ 3CAR lentivirus was successfully packaged with an average titer of 5×10 6 TU/ml. Western blotting showed that a protein band of ap- proximate 58 000 molecular weight was observed in EGFRvⅢ/3CAR-T cells but absent in untransfected T cells. Flow cytometry indi- cated the average transduction efficiency of EGFRvⅢ/3CAR was 52.3%. 51 Cr release assay showed that the specific killing effect of EGFRvⅢ/3CAR-T cells was positively correlated with E/T ratio (E∶T=4∶1, 8∶1, 16∶1, 32∶1). ELISA showed that cytokine IFN-γ se- cretion was (1 836±148.2) pg/ml, which was significantly different from that of NT T and GFP + T cells (P<0.01). The specific killing ac- tivity of EGFRvⅢ/3CAR-T cells and IFN-γ secretion were both dependent on the expression level of EGFRvⅢ in U87 cells. The tu-mor growth monitoring results showed that the tumor volume of EGFRvⅢ/3CAR-T cell group was significantly different from that of GFP + T cell group and PBS group around 3 weeks after injection (P<0.01). Conclusion: EGFRvⅢ/3CAR-T cells demonstrated specific antitumor effectagainstEGFRvⅢ + U87cellsbothinvitro and in vivo, providing basis for immunotherapyofgliomainfuture clinical use.
2018, 25(4):340-345.
DOI: 10.3872/j.issn.1007-385X.2018.04.004
Abstract:
Objective: To explore the inhibitive effect of asiatic acid (AA) on paclitaxel (PTX)-resistant glioma cells and its possible mechanism. Methods: The effects of AA on the proliferation and apoptosis of glioblastoma U87MG cells were detected by CCK-8 as- say, Real-time quantitative polymerase chain reaction (qPCR) and Western blotting. The drug-resistant glioma cell line PR-U87MG was established by culturing the cells in concentration-increasing PTX. With U87MG cells as control, the PTX-resistance of PR-U87MG cells was confirmed using CCK-8 assay, and the mRNA and protein levels of MDR1 and LRP were measured with qPCR and western blotting. PR-U87MG cells were treated with AA, PTX or AA+PTX, and then the cell viability and apoptosis of each group were mea- sured with CCK-8 assay, qPCR and Western blotting. Results: PTX-resistant PR-U87MG cell line was successfully established. AA in- hibited the viability of U87MG and PR-U87MG cells in a dose-dependent manner (P<0.01) and significantly promoted their apoptosis (P<0.01). Compared with the group treated with AA or PTX alone, the group treated with the combination of AA and PTX had signifi- cantly decreased protein levels of PARP1 (P<0.01), drug-resistant related proteins (Pgp-1 and LRP [lung resistance protein], all P< 0.01), and markedly increased caspase 3 (P<0.01). Conclusion: AA could effectively enhance the sensitivity of U87MG cells to PTX, and the mechanism may be related to the suppressed expression of drug efflux-associated proteins Pgp-1 and LRP.
Objective: To explore the inhibitive effect of asiatic acid (AA) on paclitaxel (PTX)-resistant glioma cells and its possible mechanism. Methods: The effects of AA on the proliferation and apoptosis of glioblastoma U87MG cells were detected by CCK-8 as- say, Real-time quantitative polymerase chain reaction (qPCR) and Western blotting. The drug-resistant glioma cell line PR-U87MG was established by culturing the cells in concentration-increasing PTX. With U87MG cells as control, the PTX-resistance of PR-U87MG cells was confirmed using CCK-8 assay, and the mRNA and protein levels of MDR1 and LRP were measured with qPCR and western blotting. PR-U87MG cells were treated with AA, PTX or AA+PTX, and then the cell viability and apoptosis of each group were mea- sured with CCK-8 assay, qPCR and Western blotting. Results: PTX-resistant PR-U87MG cell line was successfully established. AA in- hibited the viability of U87MG and PR-U87MG cells in a dose-dependent manner (P<0.01) and significantly promoted their apoptosis (P<0.01). Compared with the group treated with AA or PTX alone, the group treated with the combination of AA and PTX had signifi- cantly decreased protein levels of PARP1 (P<0.01), drug-resistant related proteins (Pgp-1 and LRP [lung resistance protein], all P< 0.01), and markedly increased caspase 3 (P<0.01). Conclusion: AA could effectively enhance the sensitivity of U87MG cells to PTX, and the mechanism may be related to the suppressed expression of drug efflux-associated proteins Pgp-1 and LRP.
2018, 25(4):346-350.
DOI: 10.3872/j.issn.1007-385X.2018.04.005
Abstract:
Objective: To detect the expression of zinc transporter 1 (ZnT1) gene in glioma tissue, and to explore its effect on the prolif- eration, migration and invasion of U87 cells. Methods: From October 2015 to January 2017, 20 patients with glioma, who received no chemoradiotherapy before operation, were collected from Department of Neurosurgery, the First Affiliated Hospital of China Medical University. The protein and mRNA content of ZnT1 in glioma tissues and adjacent tissues were detected by Western blotting and Real- time PCR, respectively. ZnT1 and si-ZnT1 plasmids were transfected into glioma U87 cell line respectively to construct ZnT1 over-ex- pression U87 cell line and ZnT1 knockdown U87 cell line. The effects of ZnT1 on proliferation, migration and invasion of U87 cells were detected by MTT and transwell assay. Results: Both mRNA and protein expressions of ZnT1 in glioma tissues was significantly higher than those in adjacent tissues (all P<0.05). U87 cell lines with ZnT1 over-expression and knockdown were successfully construct- ed. Compared with the control group and empty plasmid control group, the proliferation (0.54±0.01 vs 0.45±0.04, 0.43±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 over-expression were significantly increased at 12 h after transfection; how- ever, the proliferation (0.37±0.03 vs 0.45±0.01, 0.44±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 knock- down were decreased significantly. Conclusion: ZnT1 was highly expressed in glioma tissues, and promoted the proliferation, migra- tion and invasion of glioma U87 cells.
Objective: To detect the expression of zinc transporter 1 (ZnT1) gene in glioma tissue, and to explore its effect on the prolif- eration, migration and invasion of U87 cells. Methods: From October 2015 to January 2017, 20 patients with glioma, who received no chemoradiotherapy before operation, were collected from Department of Neurosurgery, the First Affiliated Hospital of China Medical University. The protein and mRNA content of ZnT1 in glioma tissues and adjacent tissues were detected by Western blotting and Real- time PCR, respectively. ZnT1 and si-ZnT1 plasmids were transfected into glioma U87 cell line respectively to construct ZnT1 over-ex- pression U87 cell line and ZnT1 knockdown U87 cell line. The effects of ZnT1 on proliferation, migration and invasion of U87 cells were detected by MTT and transwell assay. Results: Both mRNA and protein expressions of ZnT1 in glioma tissues was significantly higher than those in adjacent tissues (all P<0.05). U87 cell lines with ZnT1 over-expression and knockdown were successfully construct- ed. Compared with the control group and empty plasmid control group, the proliferation (0.54±0.01 vs 0.45±0.04, 0.43±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 over-expression were significantly increased at 12 h after transfection; how- ever, the proliferation (0.37±0.03 vs 0.45±0.01, 0.44±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 knock- down were decreased significantly. Conclusion: ZnT1 was highly expressed in glioma tissues, and promoted the proliferation, migra- tion and invasion of glioma U87 cells.
2018, 25(4):351-356.
DOI: 10.3872/j.issn.1007-385X.2018.04.006
Abstract:
Objective: To evaluate the expression level of FOXD1 in glioma tissues of different grades, and to investigate the correla- tion between the expression of FOXD1 and the prognosis of glioma patients. Methods: The tumor tissues were collected from 40 glio- ma patients, who received surgical treatment in the neurosurgery department of the First Hospital of China Medical University from September 2014 to February 2015; Seven non-tumor tissues obtained from patients underwent internal decompression for traumatic brain injury were used as controls. The FOXD1 expression in glioma and non-tumor brain tissues was analyzed by qRT-PCR and IHC, and the correlations between clinical pathological features of glioma patients and FOXD1 expression level were analyzed. Furthermore, the Kaplan-Meier method was used to analyze the relationship between FOXD1 expression and survival time of patients. In addition, the expression of FOXD1 in glioma tissues and its relationship with patients’prognosis were confirmed by the data from GEO (GSE4290, GSE2223) and Rembrandt database. Results: qRT-PCR showed that the FOXD1 mRNA expression in glioma tissues of WHO grade IV was significantly higher than that of non-tumor brain tissues and glioma tissues of WHO grade II (P<0.01). German im- munohistochemical score (GIS) was used to evaluate the immunohistochemical staining intensity, and the relationship between FOXD1 expression and clinical pathological features was analyzed. The results showed that FOXD1 in glioma tissues was related to WHO pha- thological grade level (χ 2 =11.73，P<0.01). There was statistically significant difference between the survival time of FOXD1 high ex- pression group and FOXD1 low expression group (P=0.043). The data from GEO data base (GSE4290, GSE2223) and Rembrandt data- sets showed that glioma tissues have a higher FOXD1 mRNA expression level than normal brain tissues, and the elevated expression of FOXD1 mRNA was negatively associated with the survival time of glioma patients. Conclusion: FOXD1 was highly expressed in glio- ma tissues, and the expression level of FOXD1 was increased as the pathological grade increases. The elevated expression of FOXD1 was related with the poor survival of glioma patients.
Objective: To evaluate the expression level of FOXD1 in glioma tissues of different grades, and to investigate the correla- tion between the expression of FOXD1 and the prognosis of glioma patients. Methods: The tumor tissues were collected from 40 glio- ma patients, who received surgical treatment in the neurosurgery department of the First Hospital of China Medical University from September 2014 to February 2015; Seven non-tumor tissues obtained from patients underwent internal decompression for traumatic brain injury were used as controls. The FOXD1 expression in glioma and non-tumor brain tissues was analyzed by qRT-PCR and IHC, and the correlations between clinical pathological features of glioma patients and FOXD1 expression level were analyzed. Furthermore, the Kaplan-Meier method was used to analyze the relationship between FOXD1 expression and survival time of patients. In addition, the expression of FOXD1 in glioma tissues and its relationship with patients’prognosis were confirmed by the data from GEO (GSE4290, GSE2223) and Rembrandt database. Results: qRT-PCR showed that the FOXD1 mRNA expression in glioma tissues of WHO grade IV was significantly higher than that of non-tumor brain tissues and glioma tissues of WHO grade II (P<0.01). German im- munohistochemical score (GIS) was used to evaluate the immunohistochemical staining intensity, and the relationship between FOXD1 expression and clinical pathological features was analyzed. The results showed that FOXD1 in glioma tissues was related to WHO pha- thological grade level (χ 2 =11.73，P<0.01). There was statistically significant difference between the survival time of FOXD1 high ex- pression group and FOXD1 low expression group (P=0.043). The data from GEO data base (GSE4290, GSE2223) and Rembrandt data- sets showed that glioma tissues have a higher FOXD1 mRNA expression level than normal brain tissues, and the elevated expression of FOXD1 mRNA was negatively associated with the survival time of glioma patients. Conclusion: FOXD1 was highly expressed in glio- ma tissues, and the expression level of FOXD1 was increased as the pathological grade increases. The elevated expression of FOXD1 was related with the poor survival of glioma patients.
ZHANG Dongyong
,
WANG Yiwei
,
ZHANG Luyang
,
WANG Wei
,
LIU Qiang
,
LI Zhenhang
,
WANG Yunjie
,
QIU Bo
2018, 25(4):357-362.
DOI: 10.3872/j.issn.1007-385X.2018.04.007
Abstract:
Objective: To study the effect and possible mechanism of TGF-β2 on the invasion of glioma stem cells (GSCs). Methods: Tumor tissues of 8 patients with glioblastoma multiforme, who underwent resection at Department of Neurosurgery of the First Affiliat- ed Hospital of China Medical University during April 2016 to April 2017, were collected. The primary culture of glioma cells were con- ducted with trypsin digestion. Partial primary glioma cells were seeded into serum-free DMEM/F12 culture medium containing EGF, bFGF and B27 to obtain suspension of tumor spheres. Immunoflurenscent staining and differentiation assay were used to detect whether the tumor spheres were GSCs. TGF-β2 secretion ability of GSCs was determined by ELISA assay. After transfection of TGF-β2 siRNA, the invasion ability of glioma stem cells was determined by Transwell assay. Western blotting was used to examine the effect of TGF-β2 on expression of matrix metalloproteinases (MMP) in glioma stem cells. Results: The suspended tumor spheres were proved to be GSCs by immunofluorescent staining and differentiation assay; the tumor spheres expressed the marker of GSCs（CD133）and had the ability to multi-differentiate (glia and neuronal cells). Compared with the primary glioma cells, Glioma stem cells exerted significantly improved TGF-β2 secretion ability ([74.13±3.63] vs [46.13±2.61] pg/ml, P<0.05); and TGF-β2 silencing significantly reduced the inva- sion ability of glioma stem cells ([105.71±8.69] vs [63.67±5.93], P<0.05) and inhibited MMP-2 and MMP-9 expressions. Conclusion: TGF-β2 can promote the invasiveness of glioma stem cells through MMP-2 and MMP-9 pathway.
Objective: To study the effect and possible mechanism of TGF-β2 on the invasion of glioma stem cells (GSCs). Methods: Tumor tissues of 8 patients with glioblastoma multiforme, who underwent resection at Department of Neurosurgery of the First Affiliat- ed Hospital of China Medical University during April 2016 to April 2017, were collected. The primary culture of glioma cells were con- ducted with trypsin digestion. Partial primary glioma cells were seeded into serum-free DMEM/F12 culture medium containing EGF, bFGF and B27 to obtain suspension of tumor spheres. Immunoflurenscent staining and differentiation assay were used to detect whether the tumor spheres were GSCs. TGF-β2 secretion ability of GSCs was determined by ELISA assay. After transfection of TGF-β2 siRNA, the invasion ability of glioma stem cells was determined by Transwell assay. Western blotting was used to examine the effect of TGF-β2 on expression of matrix metalloproteinases (MMP) in glioma stem cells. Results: The suspended tumor spheres were proved to be GSCs by immunofluorescent staining and differentiation assay; the tumor spheres expressed the marker of GSCs（CD133）and had the ability to multi-differentiate (glia and neuronal cells). Compared with the primary glioma cells, Glioma stem cells exerted significantly improved TGF-β2 secretion ability ([74.13±3.63] vs [46.13±2.61] pg/ml, P<0.05); and TGF-β2 silencing significantly reduced the inva- sion ability of glioma stem cells ([105.71±8.69] vs [63.67±5.93], P<0.05) and inhibited MMP-2 and MMP-9 expressions. Conclusion: TGF-β2 can promote the invasiveness of glioma stem cells through MMP-2 and MMP-9 pathway.
2018, 25(4):363-369.
DOI: 10.3872/j.issn.1007-385X.2018.04.008
Abstract:
Objective: Toevaluatetheexpressionof6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3(PFKFB3) in malignant glioma tissues and the effects of inhibitor of PFKFB3(PFK15) on the proliferation, migration, invasion, clone formation and tumorigenesis of H4 cells. Methods: Malignant brain glioma tissues and corresponding paratumor tissues from 31 patients, who were hospitalized in Department of Neurosurgery, Ankang Hospital of Traditional Chinese Medicine during February 1, 2015 to January 31, 2016 for operative treatment, were collected for this study. Immunohistochemistry and western blotting assays were applied to detect the expression of PFKFB3 in collected tissues. PFKFB3 in H4 cells were blocked by PFK15 (1.25, 2.5, 5.0 μmol/L). The effect of PFK15 on proliferation, migration, clone formation and tumorigenesis of H4 cells were determined by MTT assay, EdU incorporation assay, wound healing assay, Transwell assay, colone formation assay and in vivo xenograft bearing nude mice model respectively. Results: Positive expression rate of PFKFB3 was significantly higher in malignant glioma tissues compared with normal adjacent tissues[(80.60±8.98)% vs (41.57±10.16)%, P<0.05]. The results of MTT assay and EdU incorporation assay indicated that PEK15 significantly inhibited the proliferation of H4 cells in a concentration dependent manner. The migration, invasion and clone formation activity of H4 cells were significantly reduced by treatment with PFK15 (all P<0.05). In tumor bearing nude mice, the tumor volume of mice treated with PFK15 was significantly smaller than that of mice from control group ([254.15±154.25] vs [801.52±224.25] mm 3 , P<0.05). Conclusion: PFKFB3 was highly expressed in malignant glioma tissues. Blocking of PFKFB3 by PFK15 significantly reduced the malignant biological behaviors and tumorigenesis of H4 cells in vitro and in vivo, which may
Objective: Toevaluatetheexpressionof6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3(PFKFB3) in malignant glioma tissues and the effects of inhibitor of PFKFB3(PFK15) on the proliferation, migration, invasion, clone formation and tumorigenesis of H4 cells. Methods: Malignant brain glioma tissues and corresponding paratumor tissues from 31 patients, who were hospitalized in Department of Neurosurgery, Ankang Hospital of Traditional Chinese Medicine during February 1, 2015 to January 31, 2016 for operative treatment, were collected for this study. Immunohistochemistry and western blotting assays were applied to detect the expression of PFKFB3 in collected tissues. PFKFB3 in H4 cells were blocked by PFK15 (1.25, 2.5, 5.0 μmol/L). The effect of PFK15 on proliferation, migration, clone formation and tumorigenesis of H4 cells were determined by MTT assay, EdU incorporation assay, wound healing assay, Transwell assay, colone formation assay and in vivo xenograft bearing nude mice model respectively. Results: Positive expression rate of PFKFB3 was significantly higher in malignant glioma tissues compared with normal adjacent tissues[(80.60±8.98)% vs (41.57±10.16)%, P<0.05]. The results of MTT assay and EdU incorporation assay indicated that PEK15 significantly inhibited the proliferation of H4 cells in a concentration dependent manner. The migration, invasion and clone formation activity of H4 cells were significantly reduced by treatment with PFK15 (all P<0.05). In tumor bearing nude mice, the tumor volume of mice treated with PFK15 was significantly smaller than that of mice from control group ([254.15±154.25] vs [801.52±224.25] mm 3 , P<0.05). Conclusion: PFKFB3 was highly expressed in malignant glioma tissues. Blocking of PFKFB3 by PFK15 significantly reduced the malignant biological behaviors and tumorigenesis of H4 cells in vitro and in vivo, which may
2018, 25(4):370-375.
DOI: 10.3872/j.issn.1007-385X.2018.04.009
Abstract:
Objective: To investigate the expression and clinical significance of lncRNAANRIL in glioma tissues. Methods: 129 cas- es of glioma tissues and 25 cases of normal brain tissues as control were collected from patients treated in Sichuan Provincial People’s Hospital from January 01, 2012 to December 30, 2016. Real-time quantitative PCR was used to detect the mRNA expression of ln- cRNAANRIL; and the relationship between lncRNAANRIL expression and sensitivity of patients to temozolomide as well as the clini- cal prognosis of glioma patients were analyzed. Results: Compared with control group, the expression of lncRNAANRIL in 129 cases of glioma tissues was significantly increased ([8.730±0.336] vs [1.090±0.137], t=9.957, P<0.01). The expression of lncRNAANRIL in WHO Ⅰ-Ⅱ grade patients was significantly lower than that of patients at grade Ⅲ-Ⅳ ([4.198±0.260] vs [10.550±0.291], t=13.03, P< 0.01). lncRNA ANRIL expression was significantly correlated with WHO stage，the sensitivity to temozolomide（TMZ）and survival status（all P<0.05）, but not associated with gender, age, KPS score and tumor size (all P>0.05). Moreover, Kaplan-Meier analysis dem- onstrated that decreased lncRNA ANRIL expression contributed to significantly longer overall survival ([29.17±0.64] vs [13.54±0.74] months, P<0.01) and recurrence-free survival time ([9.08±0.56] vs [15.88±0.83] months, P<0.01). Univariate and multivariate analysis also indicated that lncRNA ANRIL expression, WHO stage and chemosensitivity could be independent prognostic markers for glioma (P<0.05). Conclusion: Higher pathological grade of glioma patients indicates higher lncRNA ANRIL expression and shorter survival time. lncRNAANRIL is involved in the occurrence and development of glioma, and can be used as a molecular marker for the diagno- sis and prognosis of glioma.
Objective: To investigate the expression and clinical significance of lncRNAANRIL in glioma tissues. Methods: 129 cas- es of glioma tissues and 25 cases of normal brain tissues as control were collected from patients treated in Sichuan Provincial People’s Hospital from January 01, 2012 to December 30, 2016. Real-time quantitative PCR was used to detect the mRNA expression of ln- cRNAANRIL; and the relationship between lncRNAANRIL expression and sensitivity of patients to temozolomide as well as the clini- cal prognosis of glioma patients were analyzed. Results: Compared with control group, the expression of lncRNAANRIL in 129 cases of glioma tissues was significantly increased ([8.730±0.336] vs [1.090±0.137], t=9.957, P<0.01). The expression of lncRNAANRIL in WHO Ⅰ-Ⅱ grade patients was significantly lower than that of patients at grade Ⅲ-Ⅳ ([4.198±0.260] vs [10.550±0.291], t=13.03, P< 0.01). lncRNA ANRIL expression was significantly correlated with WHO stage，the sensitivity to temozolomide（TMZ）and survival status（all P<0.05）, but not associated with gender, age, KPS score and tumor size (all P>0.05). Moreover, Kaplan-Meier analysis dem- onstrated that decreased lncRNA ANRIL expression contributed to significantly longer overall survival ([29.17±0.64] vs [13.54±0.74] months, P<0.01) and recurrence-free survival time ([9.08±0.56] vs [15.88±0.83] months, P<0.01). Univariate and multivariate analysis also indicated that lncRNA ANRIL expression, WHO stage and chemosensitivity could be independent prognostic markers for glioma (P<0.05). Conclusion: Higher pathological grade of glioma patients indicates higher lncRNA ANRIL expression and shorter survival time. lncRNAANRIL is involved in the occurrence and development of glioma, and can be used as a molecular marker for the diagno- sis and prognosis of glioma.
2018, 25(4):376-381.
DOI: 10.3872/j.issn.1007-385X.2018.04.010
Abstract:
脑胶质瘤（胶质瘤）具有高发病率、高病死率、高复发率及低治愈率的特点，胶质瘤细胞的无限增殖能力和高侵袭迁移 能力是胶质瘤治疗的难点，近年来微小核糖核酸（microRNA，miRNA）的出现为研究胶质瘤的发生及侵袭迁移机制提供了新的思 路。miRNA是一类内生的、长约20~24个核苷酸的非编码小RNA，可通过与其靶基因mRNA的3’UTR区域互补结合，从而在转 录后水平调控基因的表达。多项研究证实，miR-10b在胶质瘤组织和胶质瘤患者血清中高表达，并影响患者预后情况。miR-10b 可能通过影响其靶基因如PTEN、CDKN、P53、HOXD10等的表达，参与胶质瘤细胞的增殖、侵袭和迁移等过程。深入研究miR- 10b对胶质瘤的调控作用及其机制，对该病的诊断、治疗和预后评估等均具有重要意义。
脑胶质瘤（胶质瘤）具有高发病率、高病死率、高复发率及低治愈率的特点，胶质瘤细胞的无限增殖能力和高侵袭迁移 能力是胶质瘤治疗的难点，近年来微小核糖核酸（microRNA，miRNA）的出现为研究胶质瘤的发生及侵袭迁移机制提供了新的思 路。miRNA是一类内生的、长约20~24个核苷酸的非编码小RNA，可通过与其靶基因mRNA的3’UTR区域互补结合，从而在转 录后水平调控基因的表达。多项研究证实，miR-10b在胶质瘤组织和胶质瘤患者血清中高表达，并影响患者预后情况。miR-10b 可能通过影响其靶基因如PTEN、CDKN、P53、HOXD10等的表达，参与胶质瘤细胞的增殖、侵袭和迁移等过程。深入研究miR- 10b对胶质瘤的调控作用及其机制，对该病的诊断、治疗和预后评估等均具有重要意义。
2018, 25(4):382-388.
DOI: 10.3872/j.issn.1007-385X.2018.04.011
Abstract:
Objective: To quantify the expression of growth arrest and DNA damage inducible protein 45g (GADD45g) gene in the bone marrow samples of patients with AML (acute myeloid leukemia) and in AML cell lines, as well as to study the correlation between the GADD45g expression and prognostic outcome in patients with AML and investigate the role of GADD45g over-expression in prolif- eration, apoptosis, senescence, differentiation, cell cycle arrest, and drug sensitivity in AML cell lines. Methods: In the study, a total of 27 cases of bone marrow specimens were selected from patients initially diagnosed as AML in Hospital of Blood Diseases affiliated to Chinese Academy of Medical Sciences from January 2013 to December 2016. mRNA and protein expression levels of GADD45g in BMMNCs (Bone marrow mononuclear cells) from patients with AML and healthy donors and in AML cell lines were quantified by quantitative real-time PCR and Western blotting. The correlation between GADD45g expression and overall survival (OS), coupled with event-free survival (EFS) in patients withAML was analyzed in two gene expression datasets (GSE10358, GSE425-GPL317). Len- tiviral vectors over-expressing GADD45g were constructed and transfected into AML cell lines (U937, THP-1 and Molm-13 cell lines). The role of GADD45g over-expression in cell proliferation, colony formation, senescence, apoptosis, cell cycle arrest, differentiation and drug sensitivity of U937, THP-1 and Molm-13 cells were detected by cell counting, colony-forming assay, β-galactosidase staining and flow cytometric analysis of Annexin V/7AAD staining, PI staining and so on, respectively. Results: Expression of GADD45g was dramatically down-regulated in BMMNCs in AML patients and AML cell lines compared to that from healthy donors (all P<0.01). The OS (P<0.05) and EFS (P<0.05) of AML patients with low GADD45g expression were significantly shorter that those of AML patients with higher GADD45g level. Enforced expression of GADD45g could inhibit cell growth and colony formation, promote senescence and apoptosis, induce cell cycle arrest and differentiation and enhance drug sensitivity in AML cell lines (P<0.05 or P<0.01). Conclu- sion: GADD45g expression was remarkably silenced in marrow tissues of patients with AML and AML cell lines; it showed remarkable and all-around inhibiting effect onAML cell lines, indicating that GADD45g expression has prognostic value inAML.
Objective: To quantify the expression of growth arrest and DNA damage inducible protein 45g (GADD45g) gene in the bone marrow samples of patients with AML (acute myeloid leukemia) and in AML cell lines, as well as to study the correlation between the GADD45g expression and prognostic outcome in patients with AML and investigate the role of GADD45g over-expression in prolif- eration, apoptosis, senescence, differentiation, cell cycle arrest, and drug sensitivity in AML cell lines. Methods: In the study, a total of 27 cases of bone marrow specimens were selected from patients initially diagnosed as AML in Hospital of Blood Diseases affiliated to Chinese Academy of Medical Sciences from January 2013 to December 2016. mRNA and protein expression levels of GADD45g in BMMNCs (Bone marrow mononuclear cells) from patients with AML and healthy donors and in AML cell lines were quantified by quantitative real-time PCR and Western blotting. The correlation between GADD45g expression and overall survival (OS), coupled with event-free survival (EFS) in patients withAML was analyzed in two gene expression datasets (GSE10358, GSE425-GPL317). Len- tiviral vectors over-expressing GADD45g were constructed and transfected into AML cell lines (U937, THP-1 and Molm-13 cell lines). The role of GADD45g over-expression in cell proliferation, colony formation, senescence, apoptosis, cell cycle arrest, differentiation and drug sensitivity of U937, THP-1 and Molm-13 cells were detected by cell counting, colony-forming assay, β-galactosidase staining and flow cytometric analysis of Annexin V/7AAD staining, PI staining and so on, respectively. Results: Expression of GADD45g was dramatically down-regulated in BMMNCs in AML patients and AML cell lines compared to that from healthy donors (all P<0.01). The OS (P<0.05) and EFS (P<0.05) of AML patients with low GADD45g expression were significantly shorter that those of AML patients with higher GADD45g level. Enforced expression of GADD45g could inhibit cell growth and colony formation, promote senescence and apoptosis, induce cell cycle arrest and differentiation and enhance drug sensitivity in AML cell lines (P<0.05 or P<0.01). Conclu- sion: GADD45g expression was remarkably silenced in marrow tissues of patients with AML and AML cell lines; it showed remarkable and all-around inhibiting effect onAML cell lines, indicating that GADD45g expression has prognostic value inAML.
LI Jian a
,
TIAN Fang b
,
JIANG Pengjun a
,
KONG Xiangtu a
,
WU Jian b
,
YIN Tingting a
,
XING Yun
,
JIN Liang
,
HAO Ruidong
,
LIU
Gentao △
,
ZHU Xuejun
2018, 25(4):389-393.
DOI: 10.3872/j.issn.1007-385X.2018.04.012
Abstract:
Objective: To establish a chimeric antigen receptor（CAR）modified T cells specifically targeting CD19 molecule (CD19- CAR-T cells) and to testify their in vitro killing effect on target cells. Methods: CD19-CAR fragments yielded by PCR were construct- ed into pCDH-GFP lentiviral vectors by molecular cloning technology. The packaged lentiviral particles were transducted into CD3 + T cells of donors. Transduction efficiency was measured by flow cytometry and PCR. The in vitro cytotoxicity of obtained CD19- CAR-T cells against CD19 + Ramos cells was tested by 7-AAD staining. Results: The amplification folds of CD3 + T cells increased to (78.8± 23.2) folds after in vitro culture for 10 days, and about (58.3±5.4)% cells expressing GFP. About (57.4±9.3)% CD19 + Ramos cells were specifically killed by the CD19-CAR-T cells in vitro at the E∶T ratio of 5∶1. Conclusion: This study successfully established an effective method for constructing and amplifying CD19-CAR-T cells in vitro, which showed profound efficiency and specific cyto- toxity against CD19 + Ramos cells. And this report might provide an experimental evidence for clinical treatment of CD19 + B cell neo- plasmas.
Objective: To establish a chimeric antigen receptor（CAR）modified T cells specifically targeting CD19 molecule (CD19- CAR-T cells) and to testify their in vitro killing effect on target cells. Methods: CD19-CAR fragments yielded by PCR were construct- ed into pCDH-GFP lentiviral vectors by molecular cloning technology. The packaged lentiviral particles were transducted into CD3 + T cells of donors. Transduction efficiency was measured by flow cytometry and PCR. The in vitro cytotoxicity of obtained CD19- CAR-T cells against CD19 + Ramos cells was tested by 7-AAD staining. Results: The amplification folds of CD3 + T cells increased to (78.8± 23.2) folds after in vitro culture for 10 days, and about (58.3±5.4)% cells expressing GFP. About (57.4±9.3)% CD19 + Ramos cells were specifically killed by the CD19-CAR-T cells in vitro at the E∶T ratio of 5∶1. Conclusion: This study successfully established an effective method for constructing and amplifying CD19-CAR-T cells in vitro, which showed profound efficiency and specific cyto- toxity against CD19 + Ramos cells. And this report might provide an experimental evidence for clinical treatment of CD19 + B cell neo- plasmas.
2018, 25(4):394-400.
DOI: 10.3872/j.issn.1007-385X.2018.04.013
Abstract:
Objective: To prepare GNS (gold nanostars) loading photosensitizer chlorin e6 (Ce6) and to investigate its photodynamic effects on lung cancer A549 cells. Methods: GNS was firstly modified by SH-PEG-NH 2 and then mixed with Ce6 and shaken overnight to prepare GNS-PEG@Ce6, which had photodynamic therapy effects. The characterization, morphology and encapsulation rate were de- tected. The difference between the phagocytosis of Ce6 and GNS-PEG@Ce6 by A549 cells were observed with a Leical TCS SP8 con- focal laser scanning microscope. MTT assay was used to examine the inhibitory effect of GNS-PEG@Ce6 on the proliferation of A549 cells while FCM was used to detect the effect of probe GNS-PEG@Ce6 on the apoptosis of A549 cells. Results: The particle size of the GNS-PEG@Ce6 was about 100 nm. The prepared GNS-PEG@Ce6 nanoparticles exhibited good dispersion and stability and the encap- sulation rate of Ce6 was about 50%. GNS-PEG@Ce6 entered the cells by endocytosis and mainly distributed in the cytoplasm; com- pared with Ce6, GNS-PEG@Ce6 could enter the cells more effectively. The proliferation-suppression effect of GNS-PEG@Ce6 on A549 cells was significantly stronger than that of Ce6 (P<0.05). The results of flow cytometry showed that the probe exhibited strong apoptotic effect on A549 cells. Conclusion: GNS, as the drug carrier, could effectively increase the Ce6 uptake efficacy in A549 cells, thus further enhancing the killing effects of Ce6 on lung cancerA549 cells.
Objective: To prepare GNS (gold nanostars) loading photosensitizer chlorin e6 (Ce6) and to investigate its photodynamic effects on lung cancer A549 cells. Methods: GNS was firstly modified by SH-PEG-NH 2 and then mixed with Ce6 and shaken overnight to prepare GNS-PEG@Ce6, which had photodynamic therapy effects. The characterization, morphology and encapsulation rate were de- tected. The difference between the phagocytosis of Ce6 and GNS-PEG@Ce6 by A549 cells were observed with a Leical TCS SP8 con- focal laser scanning microscope. MTT assay was used to examine the inhibitory effect of GNS-PEG@Ce6 on the proliferation of A549 cells while FCM was used to detect the effect of probe GNS-PEG@Ce6 on the apoptosis of A549 cells. Results: The particle size of the GNS-PEG@Ce6 was about 100 nm. The prepared GNS-PEG@Ce6 nanoparticles exhibited good dispersion and stability and the encap- sulation rate of Ce6 was about 50%. GNS-PEG@Ce6 entered the cells by endocytosis and mainly distributed in the cytoplasm; com- pared with Ce6, GNS-PEG@Ce6 could enter the cells more effectively. The proliferation-suppression effect of GNS-PEG@Ce6 on A549 cells was significantly stronger than that of Ce6 (P<0.05). The results of flow cytometry showed that the probe exhibited strong apoptotic effect on A549 cells. Conclusion: GNS, as the drug carrier, could effectively increase the Ce6 uptake efficacy in A549 cells, thus further enhancing the killing effects of Ce6 on lung cancerA549 cells.
2018, 25(4):401-406.
DOI: 10.3872/j.issn.1007-385X.2018.04.014
Abstract:
Objective: To analyze and compare the clinical efficacy and safety of dendritic cell cytokine-induced killer cells (DC- CIK) combined with palliative therapy or chemotherapy in the treatment of advanced pancreatic carcinoma. Methods: A retrospective study was carried on 50 patients with advanced pancreatic carcinoma who were hospitalized in department of oncology of Shanxi Dayi Hospital during September 2012 to February 2016. The patients were divided into four groups according to the therapy they received (palliative treatment group, palliative+DC-CIK treatment group, chemotherapy group and chemotherapy+DC-CIK treatment group); the immunological function, quality of life and survival time of patients were analyzed; and the efficacy and safety of DC-CIK cell ther- apy was also evaluated. Results: The percentages of CD8 + T cells and NKT cells in DC-CIK combined therapy groups were significant- ly improved compared with that of pre-treatment, and the percentages of CD3 + , CD8 + , NK, NKT cells were increased compared with control groups (P<0.05). The quality of life of patients was significantly improved (P<0.05), while median PFS and median OS were improved but without statistical significance (P>0.05). Conclusion: Compared with palliative therapy and chemotherapy alone, com- bined DC-CIK immunotherapy can effectively improve the cellular immunity function and quality of life in patients with advanced pan- creatic cancer. However, there was no significant extension in overall survival.
Objective: To analyze and compare the clinical efficacy and safety of dendritic cell cytokine-induced killer cells (DC- CIK) combined with palliative therapy or chemotherapy in the treatment of advanced pancreatic carcinoma. Methods: A retrospective study was carried on 50 patients with advanced pancreatic carcinoma who were hospitalized in department of oncology of Shanxi Dayi Hospital during September 2012 to February 2016. The patients were divided into four groups according to the therapy they received (palliative treatment group, palliative+DC-CIK treatment group, chemotherapy group and chemotherapy+DC-CIK treatment group); the immunological function, quality of life and survival time of patients were analyzed; and the efficacy and safety of DC-CIK cell ther- apy was also evaluated. Results: The percentages of CD8 + T cells and NKT cells in DC-CIK combined therapy groups were significant- ly improved compared with that of pre-treatment, and the percentages of CD3 + , CD8 + , NK, NKT cells were increased compared with control groups (P<0.05). The quality of life of patients was significantly improved (P<0.05), while median PFS and median OS were improved but without statistical significance (P>0.05). Conclusion: Compared with palliative therapy and chemotherapy alone, com- bined DC-CIK immunotherapy can effectively improve the cellular immunity function and quality of life in patients with advanced pan- creatic cancer. However, there was no significant extension in overall survival.
2018, 25(4):407-413.
DOI: 10.3872/j.issn.1007-385X.2018.04.015
Abstract:
Objective: To establish a method for in vitro isolation and culture of T lymphocytes from peripheral blood of cynomolgus monkeys that induced by human CD3 antibody based on the foundation of protein homology of CD3 from human, cynomolgus mon- key and porcine. Methods: The amino acid sequences of human, cynomolgus monkeys and porcine CD3 proteins were obtained from NCBI, and the sequence, homology and phylogenetic tree were analyzed by DNAMAN software. Western blotting was used to detect the expression of CD3 protein on T cell membranes from the three species. PBMCs of healthy cynomolgus were isolated and divided in- to three groups: group A was stimulated with anti-human CD3Ab alone, group B was stimulated with IL-2 alone, and group C was co- stimulated with human CD3Ab and IL-2. Cell morphology and growth status were observed under inverted microscope and the cell growth curve was plotted. Cell viability was detected by trypan blue staining and the expressions of CD3, CD4 and CD8 on T cell sur- face were detected by flow cytometry. Results: The homology of the amino acid sequence of human CD3 protein to cynomolgus mon- key and porcine were 86.9% and 65.6% respectively. The expression levels of CD3 protein on cynomolgus and porcine T cell mem- brane were 79% and 17% contrast to human, respectively. Cells of group A did not proliferate. Proliferation, viability and CD3 expres- sion [(93.8±3.6)% vs (70.3±4.7)%, P<0.01] in T cells of group C were significantly higher than those in group B. Growth curve of T cells in group C showed an S-shape, which is consistent with Logistic growth curve. T cells in group C exhibited high purity and expressed high level CD3; moreover, the CD8 + T cell took a high proportion. Conclusion: The membrane of T lymphocytes from periph- eral blood of cynomolgus can express CD3 protein that highly homological to human. Co-stimulation of human CD3Ab, IL-2 and 1% PHA can induce the proliferation and differentiation of T lymphocytes of cynomolgus, and obtain T lymphocytes with good growth sta- tus, high proliferation ability and high purity.
Objective: To establish a method for in vitro isolation and culture of T lymphocytes from peripheral blood of cynomolgus monkeys that induced by human CD3 antibody based on the foundation of protein homology of CD3 from human, cynomolgus mon- key and porcine. Methods: The amino acid sequences of human, cynomolgus monkeys and porcine CD3 proteins were obtained from NCBI, and the sequence, homology and phylogenetic tree were analyzed by DNAMAN software. Western blotting was used to detect the expression of CD3 protein on T cell membranes from the three species. PBMCs of healthy cynomolgus were isolated and divided in- to three groups: group A was stimulated with anti-human CD3Ab alone, group B was stimulated with IL-2 alone, and group C was co- stimulated with human CD3Ab and IL-2. Cell morphology and growth status were observed under inverted microscope and the cell growth curve was plotted. Cell viability was detected by trypan blue staining and the expressions of CD3, CD4 and CD8 on T cell sur- face were detected by flow cytometry. Results: The homology of the amino acid sequence of human CD3 protein to cynomolgus mon- key and porcine were 86.9% and 65.6% respectively. The expression levels of CD3 protein on cynomolgus and porcine T cell mem- brane were 79% and 17% contrast to human, respectively. Cells of group A did not proliferate. Proliferation, viability and CD3 expres- sion [(93.8±3.6)% vs (70.3±4.7)%, P<0.01] in T cells of group C were significantly higher than those in group B. Growth curve of T cells in group C showed an S-shape, which is consistent with Logistic growth curve. T cells in group C exhibited high purity and expressed high level CD3; moreover, the CD8 + T cell took a high proportion. Conclusion: The membrane of T lymphocytes from periph- eral blood of cynomolgus can express CD3 protein that highly homological to human. Co-stimulation of human CD3Ab, IL-2 and 1% PHA can induce the proliferation and differentiation of T lymphocytes of cynomolgus, and obtain T lymphocytes with good growth sta- tus, high proliferation ability and high purity.
2018, 25(4):414-420.
DOI: 10.3872/j.issn.1007-385X.2018.04.016
Abstract:
CAR-T是一种基因改造后的细胞免疫治疗手段，T细胞输入体内后可持续活化增殖，非限制性识别并杀伤肿瘤细 胞。嵌合CD19受体的CAR-T在急性淋巴细胞白血病中的平均有效率接近80%，但在实体肿瘤中却未观察到明显疗效。肿瘤抗 原识别不佳及肿瘤微环境抑制是影响疗效的主要原因，提升抗原的特异性既能增加疗效也能避免脱靶效应的发生。本文主要综 述肿瘤抗原的特点及其在CAR-T治疗实体瘤中的应用现状，重点分析神经节苷酯（disialoganglioside，GD2）、EGFRvⅢ、CEA抗原 特点及应用情况，探讨如何对抗原进行精准选择，其中基因突变后产生的新抗原最有成为特异性抗原的潜力。
CAR-T是一种基因改造后的细胞免疫治疗手段，T细胞输入体内后可持续活化增殖，非限制性识别并杀伤肿瘤细 胞。嵌合CD19受体的CAR-T在急性淋巴细胞白血病中的平均有效率接近80%，但在实体肿瘤中却未观察到明显疗效。肿瘤抗 原识别不佳及肿瘤微环境抑制是影响疗效的主要原因，提升抗原的特异性既能增加疗效也能避免脱靶效应的发生。本文主要综 述肿瘤抗原的特点及其在CAR-T治疗实体瘤中的应用现状，重点分析神经节苷酯（disialoganglioside，GD2）、EGFRvⅢ、CEA抗原 特点及应用情况，探讨如何对抗原进行精准选择，其中基因突变后产生的新抗原最有成为特异性抗原的潜力。
2018, 25(4):421-425.
DOI: 10.3872/j.issn.1007-385X.2018.04.017
Abstract:
间充质干细胞（mesenchymal stem cell，MSC）是一类具有多向分化潜能的细胞，广泛存在于成体结缔组织和器官中， 可向损伤组织和包括肿瘤的炎症部位迁移，并参与肿瘤微环境的构成，在肿瘤发生发展的各个阶段发挥重要作用。随着研究不 断深入，间充质干细胞对肿瘤影响逐步得到认识并成为当今肿瘤领域的研究热点之一。本文主要从间充质干细胞的特性、对肿 瘤的影响、对肿瘤微环境和免疫细胞的作用、参与血管生成以及相关临床试验等方面来进行综述。
间充质干细胞（mesenchymal stem cell，MSC）是一类具有多向分化潜能的细胞，广泛存在于成体结缔组织和器官中， 可向损伤组织和包括肿瘤的炎症部位迁移，并参与肿瘤微环境的构成，在肿瘤发生发展的各个阶段发挥重要作用。随着研究不 断深入，间充质干细胞对肿瘤影响逐步得到认识并成为当今肿瘤领域的研究热点之一。本文主要从间充质干细胞的特性、对肿 瘤的影响、对肿瘤微环境和免疫细胞的作用、参与血管生成以及相关临床试验等方面来进行综述。
2018, 25(4):426-430.
DOI: 10.3872/j.issn.1007-385X.2018.04.018
Abstract:
随着对肿瘤免疫逃逸机制研究的不断深入，通过阻断程序性死亡因子1（programmed cell death 1，PD-1）及其配体l （PD-1 ligand，PD-L1）构成的 PD-1/PD-L1 通路，证实了 PD-1/PD-L1 抑制剂在晚期非小细胞肺癌（non small cell lung cancer， NSCLC）患者生存的相关性，以及PD-L1作为疗效预测标志物的价值。在NSCLC的局部晚期维持治疗、二线治疗及部分一线治 疗的临床试验中，PD-1/PD-L1抑制剂治疗均获得了较好的治疗效果；PD-1/PD-L1抑制剂联合放化疗、联合细胞因子与其他免疫 抑制剂及联合细胞外调节蛋白激酶(extracel lular regulated protein kinase，ERK)通路靶向治疗也有一定获益。本文就PD-1/PD-L1 抑制剂用于NSCLC治疗现状及其影响因素作一综述。
随着对肿瘤免疫逃逸机制研究的不断深入，通过阻断程序性死亡因子1（programmed cell death 1，PD-1）及其配体l （PD-1 ligand，PD-L1）构成的 PD-1/PD-L1 通路，证实了 PD-1/PD-L1 抑制剂在晚期非小细胞肺癌（non small cell lung cancer， NSCLC）患者生存的相关性，以及PD-L1作为疗效预测标志物的价值。在NSCLC的局部晚期维持治疗、二线治疗及部分一线治 疗的临床试验中，PD-1/PD-L1抑制剂治疗均获得了较好的治疗效果；PD-1/PD-L1抑制剂联合放化疗、联合细胞因子与其他免疫 抑制剂及联合细胞外调节蛋白激酶(extracel lular regulated protein kinase，ERK)通路靶向治疗也有一定获益。本文就PD-1/PD-L1 抑制剂用于NSCLC治疗现状及其影响因素作一综述。
2018, 25(4):431-436.
DOI: 10.3872/j.issn.1007-385X.2018.04.019
Abstract:
间变性淋巴瘤激酶（anaplastic lymphoma kinase，ALK）基因重排是发生非小细胞肺癌（non-small cell lung cancer， NSCLC）的重要致癌驱动因素之一，ALK阳性的NSCLC患者易发生脑转移，而ALK靶向抑制剂相对于一线化疗对脑转移有更好 的疗效。但经第一代ALK抑制剂治疗后患者易耐药，进而出现颅内进展，第二代和第三代ALK抑制剂可增强其对中枢神经系统 的渗透性、提高其到达靶点后的结合力，对脑转移癌有较好的治疗效果。本文回顾靶向治疗在ALK阳性NSCLC脑转移患者治疗 方面取得的进展，并对目前存在的问题及未来发展方向进行探讨。
间变性淋巴瘤激酶（anaplastic lymphoma kinase，ALK）基因重排是发生非小细胞肺癌（non-small cell lung cancer， NSCLC）的重要致癌驱动因素之一，ALK阳性的NSCLC患者易发生脑转移，而ALK靶向抑制剂相对于一线化疗对脑转移有更好 的疗效。但经第一代ALK抑制剂治疗后患者易耐药，进而出现颅内进展，第二代和第三代ALK抑制剂可增强其对中枢神经系统 的渗透性、提高其到达靶点后的结合力，对脑转移癌有较好的治疗效果。本文回顾靶向治疗在ALK阳性NSCLC脑转移患者治疗 方面取得的进展，并对目前存在的问题及未来发展方向进行探讨。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.