CO2气腹及ABT-737对TAMs/胃癌细胞共培养体系VEGF表达的影响

摘要

目的:研究CO2气腹环境下胃癌细胞增殖、VEGF表达情况,及Bcl-2特异性抑制剂对共培养体系VEGF表达的影响.方法:体外诱导TAMs细胞,共培养TAMs/MKN-45细胞,体外模拟CO2气腹环境,MTT法检测胃癌细胞增殖活性,ELISA法检测细胞培养上清中VEGF浓度.结果:共培养体系分为CO2气腹组及对照组,5 mmHg、10 mmHg及15 mmHg气腹组MKN-45细胞增殖活性及VEGF表达与对照组无差异;25 mmHg气腹组细胞增殖活性及VEGF表达与对照组比较差异明显.共培养细胞对照组和CO2气腹培养组(15mmHg)分别加入ABT-737,共培养+ABT-737组VEGF表达较对照组明显降低(P=0.001).气腹共培养加ABT-737组VEGF表达明显低于不加抑制剂的同组对照(P=0.000).结论:正常腹腔镜气腹环境并无刺激肿瘤细胞过度增殖弊端.高于正常气腹压力条件可能会加重肿瘤间质微环境的缺氧状态,进一步促进肿瘤的恶性进展.肿瘤细胞与TAMs间相互作用可能通过Bcl-2-VEGF旁分泌环路来实现.%Objective:Study on proliferation of gastric cancer cells (GCCs) and expressions of VEGF under the CO2 pneumoperitoneum environment,and effect of Bcl-2 specific inhibitor on the expression of VEGF in GCCs/TAMs co-culture system.Methods:TAMs induced by PMA and IL-4 in vitro.TAMs and MKN-45 cells were co-cultured in Transwell chamber,in pseudo CO2 pneumoperitoneum environment.Proliferation activity of GCCs was detected by MTT assay,and ELISA method was used to detect VEGF concentration in the cell culture supernatant.Results:Co-culture system was divided into CO2 pneumoperitoneum group and control group.Proliferation activity of MKN-45 cell and expression of VEGF of 5mmHg,10mmHg and 15mmHg groups are no difference with the control group;25 mmHg group is opposite to the former.The cells in co-culture group and CO2 pneumoperitoneum group (15 mmHg) were added to ABT-737,VEGF expression of co-culture + ABT-737 group was significantly lower than that of in the control group(P=0.001).Co-cultured cells VEGF expression in pneumoperitoneum group was significantly lower than the control group without inhibitor (P=0.000).Conclusion:Normal laparoscopic pneumoperitoneum is no stimulation of tumor cell proliferation defects,And higher pneumoperitoneum pressure might increase the hypoxia status of tumor microenvironment,promote tumor malignant progression.The interaction between tumor cells and TAMs may be achieved by paracrine Bcl-2-VEGF loop.