巴佛洛霉素A1抑制胃癌SGC-7901细胞中VEGF-C的表达  

Bafilomycin A1 inhibits the expression of VEGF-C in gastric cancer SGC-7901 cells

作　　者：陈海姣[1] 吴淼淼 刘鹏飞[2] 张婷 朱琳 江柯炜 沈卫东[2] CHEN Haijiao;WU Miaomiao;LIU Pengfei;ZHANG Ting;ZHU Lin;JIANG Kewei;SHEN Weidong(Central Laboratory,Affiliated Jiangyin Hospital of Nantong University,Jiangyin 214400,Jiangsu Province,China;Department of Gastroenterology,Affiliated Jiangyin Hospital of Nantong University,Jiangyin 214400,Jiangsu Province,China)

机构地区：[1]南通大学附属江阴医院中心实验室,江苏江阴214400 [2]南通大学附属江阴医院消化内科,江苏江阴214400

出　　处：《肿瘤》2021年第2期91-99,共9页Tumor

基　　金：江苏省自然科学基金(编号:BK20171157);无锡市卫生健康委科技成果与适宜技术推广项目(编号:T201929)。

摘　　要：目的:研究巴佛洛霉素A1(Bafilomycin A1)对胃癌细胞株SGC-7901的增殖、迁移和血管生成的影响,并探讨可能的机制。方法:巴佛洛霉素A1(10 nmol/mL)处理SGC-7901细胞后,检测液泡-ATP酶(vacular-ATPases,V-ATPases)活性及培养液pH值,并分别用CCK-8法检测对细胞增殖的影响,Transwell小室实验检测对细胞迁移能力的影响,ELISA法检测对SGC-7901细胞分泌血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C)蛋白能力的影响,实时荧光定量PCR和蛋白质印迹法检测对细胞中VEGF-C mRNA和蛋白表达水平的影响。收集巴佛洛霉素A1处理后SGC-7901细胞的培养上清液与人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)共培养,观察血管成管情况。结果:与对照组相比,用巴佛洛霉素A1处理后,SGC-7901细胞的V-ATPase活性受到抑制(P<0.05),细胞外培养液的pH值升高(P<0.05),细胞的增殖和迁移能力被抑制(P值均<0.05),分泌到细胞外的VEGF-C蛋白明显减少(P<0.05),细胞中VEGF-C mRNA和蛋白的表达水平均明显下调(P值均<0.05);HUVECs的成管能力减弱(P<0.05)。结论:巴佛洛霉素A1能够通过抑制V-ATPase的活性来调控胃癌SGC-7901细胞外的酸性微环境,进而抑制肿瘤细胞的增殖,影响VEGF-C的表达、分泌和新生血管的形成。Objective:To investigate the effects of Bafromycin A1 on the proliferation,migration and neovascularization of gastric cancer cell line SGC-7901 and explore its possible mechanism.Methods:SGC-7901cells were treated with 10 nmol/mL Bafilomycin A1.Then the expression of vacuolar-ATPases(V-ATPases)was detected by V-ATPase detection kit,and the extracellular pH was measured with a pH meter.The proliferation of SGC-7901 cells was detected by CCK-8 assay.The migration ability of SGC-7901 cells was detected by Transwell chamber method.The expression levels of vascular endothelial growth factor-C(VEGF-C)mRNA and protein were detected by real-time fluorescent quantitative PCR and Weatern blotting,respectively.The expression of VEGF-C in the supernatant was detected by ELISA kit.Human umbilical vein endothelial cells(HUVECs)were cultured in the supernatant of SGC-7901 cells treated with different methods to observe the angiogenesis.Results:The V-ATPase activity of SGC-7901 cells was inhibited and the extracellular pH value was increased(both P<0.05)after treated with 10 nmol/L Bafromycin A1.The proliferation and migration abilities of SGC-901 cells in 10 nmol/L Bafromycin A1 treatment groups were significantly decreased(both P<0.05).The expression levels of VEGF-C mRNA and protein were significantly decreased as compared with the control group(both P<0.05).Compared with the control group,the secretion of VEGF-C was decreased(P<0.05),and the ability of cell tube formation was weakened(P<0.05).Conclusion:These results suggest that BafilomycinA1 can regulate the extracellular acidic microenvironment of gastric cancer SGC-7901 cells by inhibiting the activity of V-ATPase,then inhibit the proliferation of tumor cells,affect the expression and secretion of VEGF-C,and the formation of new blood vessels.

关 键 词：胃肿瘤 血管内皮生长因子C H(+)K(+)交换ATP酶 巴佛洛霉素A1 新生血管化 病理性 

分 类 号：R735.2[医药卫生—肿瘤]