幽门螺杆菌感染诱导微小RNA?181c对胃癌细胞增殖和侵袭的影响

目的探讨幽门螺杆菌(Hp)感染诱导微小RNA-181c(miR-181c)对胃癌细胞增殖、侵袭和迁移的影响。方法采用实时定量PCR(QPCR)检测Hp感染人正常胃黏膜GES-1细胞和胃癌细胞(BGC-823、SGC-7901、AGS)的miR-181c水平。采用脂质体向SGC-7901细胞转染miR-181c抑制物(Inhibitor组)或阴性对照序列(NC组),另取未转染的细胞为对照组;转染48 h后采用QPCR检测miR-181c水平,活细胞计数(CCK-8)法、划痕实验和Transwell小室实验检测细胞增殖活力、划痕愈合率和穿膜细胞数以评估细胞增殖、迁移和侵袭能力,QPCR和Western blotting检测Bcl-2、基质金属蛋白酶(MMP)-9和神经型钙黏蛋白(N-cad)水平。结果QPCR检测结果显示Hp感染后各细胞的miR-181水平较感染前均升高(P<0.05),GES-1、BGC-823、SGC-7901和AGS细胞Hp感染后的miR-181c水平分别为感染前的4.37、1.63、3.25和2.09倍。与对照组和NC组相比,Inhibitor组SGC-7901细胞的miR-181c水平和转染48、72 h后的增殖活力降低(P<0.05);Inhibitor组SGC-7901细胞的划痕愈合率和穿膜细胞数量分别为(21.679±3.762)%和(128.056±21.463)个,低于对照组的(65.004±2.309)%和(325.07±34.082)个及NC组的(65.675±2.914)%和(328.035±31.391)个,差异有统计学意义(P<0.05);与对照组和NC组相比,Inhibitor组的Bcl-2、MMP-9和N-cad水平均降低(P<0.05)。对照组和NC组上述指标的差异无统计学意义(P>0.05)。结论Hp感染可升高胃癌细胞的miR-181c水平,下调该miR-181c水平对增殖、侵袭和迁移具有明显抑制作用,为Hp感染的胃癌治疗提供了新的靶点。

Effect of Helicobacter pylori infection induced microRNA-181c on the proliferation and invasion of gastric cancer cells

Objective To investigate the effect of microRNA-181c(miR-181c)induced by Helicobacter pylori(HP)infection on the proliferation,invasion and migration of gastric cancer cells.Methods The miR-181c levels of GES-1 cells and gastric cancer cells(BGC-823,SGC-7901 and AGS)were detected by real-time quantitative PCR(QPCR).SGC-7901 cells were transfected with miR-181c inhibitor(Inhibitor group)or negative control sequence(NC group)by liposome,and cells without transfection were taken as Control group.After 48 hours of transfection,the miR-181c level was detected by QPCR.CCK-8,scratch test and Transwell chamber test were used to evaluate the proliferative activity,scratch healing rate and number of transmembrane cells as to assess abilities of cell proliferation,migration and invasion.Levels of Bcl-2,matrix metalloproteinase(MMP)-9 and N-cadherinn(N-cad)were detected by QPCR and Western blotting.Results The QPCR showed that the levels of miR-181c in cells infected with HP were higher than those before infection(P<0.05).The levels of miR-181c in GES-1,BGC-823,SGC-7901 and AGS cells infected with HP were 4.37,1.63,3.25 and 2.09 times higher than those before infection,respectively.Compared with Control group and NC group,miR-181c level and proliferation activity at 48 and 72 hours after transfection of SGC-7901 cells in Inhibitor group decreased(P<0.05).The scratch healing rate and number of membrane-penetrating cells in Inhibitor group were(21.679±3.762)%and 128.056±21.463,lower than(65.004±2.309)%and 325.07±34.082 in Control group and(65.675±2.914)%and 328.035±31.391 in NC group(P<0.05).Compared with the Control group and NC group,the levels of Bcl-2,MMP-9 and N-cad decreased in Inhibitor group(P<0.05).There was no significant difference between the Control group and NC group(P>0.05).Conclusion HP infection can increase the level of miR-181c in gastric cancer cells,and down-regulating the level of miR-181c can significantly inhibit the proliferation,invasion and migration of gastric cancer cells,which provides a new target for the treatment of HP infected gastric cancer.