整合素连接激酶对幽门螺杆菌所致细胞凋亡的影响

Role of integrin-linked kinase in the effect of Helicobacter pylori on gastric cancer cell apoptosis

目的探讨整合素连接激酶对幽门螺杆菌(Hp)致胃癌细胞MGC-803凋亡的影响。方法实验分为空白对照组(胃癌细胞MGC-803正常条件培养),激动剂作用组(MGC803细胞与0.4μmol/L的整合素连接激酶激动剂LPTP共培养),Hp作用组(MGC803细胞与Hp(MOI值为100∶1)共培养)和激动剂加Hp作用组(MGC803细胞与0.4μmol/L的LPTP共培养1h,与Hp(MOI值为100∶1)共培养)。24h后流式细胞术检测细胞凋亡率,Western blot检测细胞凋亡相关蛋白PARP、Casepase-3的表达。结果激动剂作用组、Hp作用组、激动剂加Hp作用组细胞凋亡率分别为(23.45±1.484 92)%、(30.7±3.111 27)%、(16.15±1.202 08)%,与空白对照组(10.1±0.707 11)%比较,差异均有统计学意义(t值分别为-11.478,-9.131,-6.135,P<0.05);激动剂加Hp作用组与Hp作用组比较差异有统计学意义(t=6.169,P<0.05)。激动剂作用组、Hp作用组、激动剂加Hp作用组PARP蛋白相对表达量分别为0.89±0.57、1.245±0.21、0.87±0.057,与空白对照组0.6±0.06比较差异均有统计学意义(t值分别为-4.734,-13.492,-4.401,P<0.05);Casepase-3蛋白表达分别为0.78±0.35、1.32±0.04、0.87±0.45;与空白对照组0.47±0.07比较差异有统计学意义(t值分别为-5.635,-14.577,-8.037,P<0.05)。结论整合素连接激酶在Hp感染MGC-803细胞中起抗凋亡的作用。

Objective To examine the effect of integrin-linked kinase on apoptosis of the MGC-803 gastric cancer cell line induced by H.pylori. Methods MGC-803 gastric cancer cells were divided into a blank control group（cultured under normal conditions）,cells cultured in the presence of LPTP（MGC803 cells cultured in the presence of 0.4μmol/L LPTP,an integrin-linked kinase agonist）,cells co-cultured with H.pylori（MGC803 cells cultured with H.pylori（MOI=100：1））,cells co-cultured with H.pylori in the presence of LPTP（MGC803 cells cultured with H.pylori for1 hin the presence of 0.4μmol/L LPTP（MOI= 100：1）co-culture）.After 24 h,the rate of apoptosis was detected using flow cytometry,and the level of PARP and Casepase-3 expression was detected using Western blotting. ResultsThe rate of apoptosis was（23.45.±1.48492）%in cells cultured in the presence of LPTP,（30.7±3.11127）%in cells co-cultured with H.pylori,（16.15±1.20208）%in cells co-cultured with H.pylori in the presence of LPTP.The rate of apoptosis differed significantly from that（10.1±0.70711%）in the blank control group（t=-11.478,-9.131,and-6.135;P〈0.05）.The rate of apoptosis differed significantly in cells co-cultured with H.pylori in the presence of LPTP and cells co-cultured with H.pylori（t=6.169,P〈0.05）.The level of PARP protein expression was 0.89±0.57 in cells cultured in the presence of LPTP,1.245±0.21 in cells co-cultured with H.pylori,and 0.87±0.057 in cells co-cultured with H.pylori in the presence of LPTP.The level of PARP protein expression differed significantly from that（0.6±0.06）in the blank control group（t=-4.734,-13.492,and-4.401;P〈0.05）.The level of Casepase-3 protein expression was 0.78±0.35 in cells cultured in the presence of LPTP,1.32±0.04 in cells co-cultured with H.pylori,and 0.87±0.45 in cells co-cultured with H.pylori in the presence of LPTP.The level of Casepase-3 protein expression differed significantly from that（0.47±0.07）in the blank control group（t=-5.663,-14.577,and-8.037;P〈0.05）. Conclusion Integrin-linked kinase plays an anti-aging role in MGC-803 cells infected with H.pylori.