高表达TIP30对胃癌细胞SGC-7901增殖和凋亡影响及机制  

Effect of overexpressed TIP30 on proliferation and apoptosis of human gastric cancer cell line SGC-7901 and its mechanism

作　　者：刘庆伟[1] 李勇[1] 檀碧波[1] 范立侨[1] 赵群[1] 苑新宇 LIU Qing-wei;LI Yong;TAN Bi-bo;FAN Li-qiao;ZHAO Qun;YUAN Xin-yu(Third Department of General Surgery,Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北医科大学第四医院外三科,河北石家庄050011

出　　处：《中华肿瘤防治杂志》2021年第14期1080-1085,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：河北省科技支撑项目(172777119);河北省卫生厅重点科技研究项目(20190701)。

摘　　要：目的观察Tat结合蛋白30(TIP30)在胃癌组织及细胞中的表达,探讨其对胃癌细胞增殖、凋亡的影响和作用机制。方法选取2018-02-01-2019-02-01河北医科大学第四医院80例胃癌患者癌及癌旁组织(距癌周>5cm)。人胃癌细胞株MGC-803、SGC-7901、BGC-823及正常胃黏膜细胞GES-1均购自中国科学院上海细胞库。采用qRT-PCR RT-PCR检测癌及癌旁组织TIP30 mRNA表达情况,分析其与临床病理参数的关系;检测TIP30在胃癌细胞株MGC-803、SGC-7901、BGC-823及正常胃黏膜细胞GES-1的表达;采用pcDNA3.1-TIP30质粒转染SGC-7901细胞使其高表达TIP30;CCK-8法检测细胞的增殖变化;流式细胞仪检测其凋亡率;蛋白质印迹法检测Ki-67、PCNA、cleaved Caspase-3、PI3K、p-AKT和AKT蛋白表达的改变;酶标仪检测Caspase酶活性。结果TIP30 mRNA在胃癌组织(0.15±0.09)相对表达水平较癌旁组织(0.43±0.19)降低,差异有统计学意义(t=-14.228,P<0.001),且其表达与肿瘤浸润深度、有无淋巴结转移及TNM分期有关,t值分别为8.265、6.826和3.801,均P<0.001;胃癌细胞株MGC-803(0.75±0.11)、SGC-7901(0.36±0.12)和BGC-823(0.55±0.17)中的TIP30蛋白表达水平均低于GES-1细胞(1.24±0.16),差异有统计学意义,F=76.89,P<0.05;高表达TIP30后胃癌细胞株SGC-7901的增殖能力降低,F=89.12,P<0.05;凋亡率较阴性对照组明显升高,F=10^(7).10,P<0.05;高表达TIP30后可下调Ki-67、PCNA、PI3K和p-AKT蛋白的表达,F值分别为197.71、142.70、215.60和189.83,均P<0.05,上调cleaved Caspase-3蛋白的表达(F=267.50,P<0.05),AKT蛋白表达无明显变化(F=56.78,P=0.84),且Caspase-3酶的活性明显增加,F=117.39,P<0.05。结论 TIP30在胃癌组织和胃癌细胞中呈低表达,高表达TIP30可能抑制PI3K/AKT信号通路激活,促进凋亡,抑制胃癌细胞SGC-7901的增殖。Objective To investigate the expression of TIP30 in gastric cancer tissues and cell lines as well as its effect on proliferation and apoptosis of human gastric cancer cells SGC-7901 and its possible mechanism.Methods The expression of TIP30 mRNA in 80 cases of gastric cancer tissues and their adjacent tissues was detected by quantitative real-time polymerase chain reaction(qRT-PCR).The re1 ationship between TIP30 expression and clinicopathological factors of gastric carcinoma was analyzed.The expression of TIP30 was detected in gastric cancer cells MGC-803,SGC-7901,BGC-823 and the normal gastric mucosa cell line GES-1.Previously constructed pcDNA3.1-TIP30 plasmid were transiently transfected into SGC-7901 cells.The proliferation of cells were detected by using CCK-8 assay.Flow cytometry was used to detect the cell apoptosis.Western blot was used to examine the changes of concentration of Ki-67,PCNA,cleaved Caspase-3,PI3 K,p-AKT,AKT proteins.The activity of Caspase-3 enzyme was detected by spectrophotometry.Results The relative expression of TIP30 in gastric carcinoma tissues was lower than that in the adjacent tissues,the difference was statistically significant(t=-14.228,P<0.001).Its expression was related to depth of invasion,lymph node metastasis and TNM clinical stage(t=8.265,6.826,3.801,all P<0.001).The relative expression of TIP30 in gastric cancer cells SGC-7901 was lower than that in GES-1 cells,the difference was statistically significant(F=76.89,P<0.05).As compared with negative control group,overexpressed TIP30 inhibited the proliferation of SGC-7901(F=89.12,P<0.05);overexpressed TIP30 promoted the apoptosis of SGC-7901(F=107.10,P<0.05).The expression level of Ki-67,PCNA,PI3 K,p-AKT in TIP30 overexpressing cells group were significantly decreased,the difference was statistically significant(F=197.71,142.70,215.60,189.83,all P<0.05),meanwhile the expression level of cleaved Caspase-3 was significantly higher than before,the difference was statistically significant(F=267.50,P<0.05).The expression of AKT

关 键 词：胃癌 TIP30 细胞增殖 细胞凋亡 PI3K/AKT 

分 类 号：R735.2[医药卫生—肿瘤]