整合素β1亚基对幽门螺杆菌感染胃上皮细胞GES-1凋亡及增殖的影响

Effect of integrin β1 on gastric epithelial cells (GES-1) infected with Helicobacter pylori

目的分析幽门螺杆菌感染胃上皮细胞过程中整合素β1亚基表达的变化,观察整合素β1对幽门螺杆菌感染胃上皮细胞GES-1凋亡及增殖的影响。方法将幽门螺杆菌与胃上皮细胞GES-1共培养,采用流式细胞术检测整合素β1的表达量变化;应用iRNA技术降低胃上皮细胞GES-1中β1亚基的表达;幽门螺杆菌感染胃上皮细胞及iRNA降低β1的表达后,采用流式细胞术检测细胞的凋亡,采用CCK-8法检测细胞增殖情况。结果幽门螺杆菌感染胃上皮细胞12、24、48h后空白对照组凋亡率分别为(10.80±0.71)%、(12.23±0.06)%和(12.30±2.12)%,Hp感染组凋亡率分别为(24.20±0.14)%、(30.05±2.47)%和(26.40±1.91)%,差异均有统计学意义(t值分别为-26.28、-7.701和-12.855,P<0.05);与幽门螺杆菌共培养6、24、48h胃上皮细胞增殖受到抑制,抑制率分别为(35.00±3.22)%、(40.96±2.45)%和(8.00±3.33)%;幽门螺杆菌与胃上皮细胞GES-1共培养24、48h流式细胞术检测整合素β1表达,空白对照组平均荧光强度(MFI)分别为(1616.33±24.70)和(1834.67±17.01),Hp感染组为(1484.00±60.89)和(1376.00±14.11),差异有统计学意义(t值分别为3.488和35.95,P<0.05);应用iRNA技术降低胃上皮细胞GES-1整合素β1亚基表达24、48h,对照组凋亡率分别为(18.75±3.59)%和(39.75±3.68)%,iRNA组凋亡率分别为(26.25±4.11)%和(55.30±5.57)%,差异有统计学意义(t值为-2.746和-4.656,P<0.05);胃上皮细胞中β1亚基的表达降低后细胞增殖受到抑制,24、36、48、72h抑制率分别为(6.16±1.07)%、(17.15±2.76)%、(10.84±1.34)%和(21.87±1.72)%。结论幽门螺杆菌能降低胃上皮细胞GES-1整合素β1亚基的表达,而且可能通过降低整合素β1亚基的表达促进上皮细胞GES-1的凋亡并抑制其增殖。

Objectives To analyze the expression of the integrin β1subunit in gastric epithelial cells（GES-1）co-cultured with H.pylori and to explore the effects of β1in those cells. Methods Small interference RNA was used to decrease the expression of integrin β1in GES-1cells.The expression of integrin β1on GES-1and the rate of cell apoptosis were detected with flow cytometry.CCK-8was used to test cell proliferation by detecting absorbance of 450 nm. Results GES-1cells co-cultured with H.pylori for 12 h,24h,and 48 h.The rate of apoptosis in blank controls was 10.80 0.71%,12.23 0.06%,and 12.30 2.12%,respectively.The rate of apoptosis in cells that were positive for H.pylori was24.20 0.14%,30.05 2.47%,and 26.40 1.91%.The rate of apoptosis in cells that were positive for H.pylori was significantly higher than that in blank controls（t=26.28,7.701,and 7.701,respectively,P0.05）.GES-1cells co-cultured with H.pylori for 6h,24 h,and 48 h.The rate of inhibition of GES-1cell proliferation was 35.00 3.22%,40.96 2.45%,and 8.00 3.33%,respectively.H.pylori markedly inhibited the proliferation of GES-1cells.GES-1cells were co-cultured with H.pylori for 24 hand 48h,and the expression of integrin β1was detected with flow cytometry.The mean fluorescence intensity（MFI）in blank controls was 1616.33 24.70 and 1834.67 17.01,and the MFI in cells that were positive for H.pylori was 1484.00 60.89 and 1376.00 14.11.The level of integrin β1expression in cells that were positive for H.pylori was significantly lower than that in blank controls（t=3.488,3.488,P〈0.05）.Small interference RNA was used to decrease the expression of integrin β1in GES-1cells for 24 hand 48h.The rate of apoptosis in controls was 18.75 3.59% and 39.75 3.68%.The rate of apoptosis in cells transfected with iRNA was 26.25 4.11%and 55.30 5.57%.The rate of apoptosis in cells transfected with iRNA was significantly higher than that in the controls（t=2.746,4.656,P〈0.05）.When small iRNA was transfected for 24 h,36h,48 h,and 72 h,the rate of inhibition of cell proliferation was 6.16 1.07%,17.15 2.76%,10.84 1.34%,and 21.87 1.72%,respectively.Cell proliferation decreased with a decrease in integrin β1. Conclusion H.pylori may regulate the expression of integrin β1 in GES-1 cells.H.pylori may increase the apoptosis and decrease the proliferation of GES-1 cells by decreasing the expression of the integrin β1 subunit.