miRNA-19a在结肠癌中的表达及对结肠癌细胞增殖活性的影响分析

Analysis of Mirna-19a Expression in Colon Cancer and its Effect on Proliferation Activity of Colon Cancer Cells

目的探讨miRNA-19a在结肠癌中的表达及对结肠癌细胞增殖活性的影响,为结肠癌的诊断和治疗提供参考依据。方法选取结肠癌组织20例,结肠癌旁组织20例,结肠息肉组织20例,采用RT-PCR检测miRNA-19a在各组标本组织中的表达量。采用脂质体介导转染方法将miRNA-19a模拟物或抑制物转染到人结肠癌LoVo细胞;检测转染后miRNA-19a基因的表达;利用CCK8试剂盒检测细胞的增殖能力;TUNEL法检测miRNA-19a对细胞周期、细胞凋亡的影响;细胞侵袭小室法检测细胞的体外侵袭力。结果miRNA-19a在结肠癌组织和LoVo细胞中呈现高水平表达。各组结肠癌LoVo细胞增殖能力由高到低排列:转染miRNA-19a模拟物组、空白对照组、转染miRNA-19a抑制物组,且各组间的差异具有统计学意义(P<0.05)。转染miRNA-19a模拟物组的结肠癌LoVo细胞的细胞数在S期与G0/G1期、G2/M期比较差异显著,具有统计学意义(P<0.05);其他两组在S期与G0/G1期、G2/M期比较差异不明显,不具有统计学意义(P>0.05)。各组结肠癌LoVo细胞凋亡能力由低到高排列:转染miRNA-19a模拟物组、空白对照组、转染miRNA-19a抑制物组,且各组间的差异具有统计学意义(P<0.05)。各组结肠癌LoVo细胞体外侵袭能力由高到低排列:转染miRNA-19a模拟物组、空白对照组、转染miRNA-19a抑制物组,且各组间的差异具有统计学意义(P<0.05)。结论下调miRNA-19a的表达水平,可抑制癌细胞的增殖,使其在细胞周期S期细胞数减少,加快癌细胞凋亡进程,使癌细胞凋亡数量增多,增强癌细胞体外侵袭能力。

Objective To investigate the expression of miRNA-19a in colon cancer and its effect on the proliferation activity of colon cancer cells,so as to provide reference for future clinical diagnosis and treatment of colon cancer patients.Methods 20 cases of colon cancer tissues,20 cases of paraccolon cancer tissues and 20 cases of colon polyps were selected.The expression of miRNA-19a in each group was detected by rt-pcr.miRNA-19a analogue or inhibitor was transfected into human colon cancer LoVo cells by liposome mediated transfection.The expression of miRNA-19a gene was detected after transfection.Cell proliferation ability was detected by CCK8 kit.TUNEL assay was used to detect the effects of miRNA-19a on cell cycle and apoptosis.The invasiveness of cells in vitro was measured by cell invasion chamber method.Results miRNA-19a was highly expressed in colon cancer tissues and LoVo cells.The proliferation capacity of colon cancer LoVo cells in each group was ranked from high to low:transfected mi miRNA-19a analogue group,blank control group,transfected miRNA-19a inhibitor group,and the difference between the 2 groups was statistically significant(P<0.05).The number of colon cancer LoVo cells transfected with miRNA-19a analogue group was significantly different from that of G0/G1 and G2/M at S phase(P<0.05).There was no significant difference between the other 2 groups at S stage,G0/G1 stage,G2/M stage,and no statistical significance(P>0.05).The apoptosis ability of colon cancer LoVo cells in each group was ranked from low to high:transfected miRNA-19a analogue group,blank control group,transfected miRNA-19a inhibitor group,and the difference between each group was statistically significant(P<0.05).In vitro invasion ability of colon cancer LoVo cells in each group was ranked from high to low:transfected miRNA-19a analogue group,blank control group,transfected miRNA-19a inhibitor group,and the differences among the groups were statistically significant(P<0.05).Conclusion Down-regulating the expression level of mirna-19a can inhibit the proliferation of cancer cells,reduce the number of cells in cell cycle S,accelerate the apoptosis process of cancer cells,increase the apoptosis number of cancer cells,and enhance the invasiveness of cancer cells in vitro.