miR-195通过靶向调控趋化因子5促进胃癌细胞增殖、转移及侵袭的实验研究

MiR-195 Targeting Chemokine 5 Inhibits Proliferation,Metastasis and Invasion of Gastric Cancer Cells

目的:研究miR-195通过靶向调控趋化因子5促进胃癌细胞增殖、转移及侵袭的分子机制。方法:选取MGC803及NCI-N87细胞,根据转染不同分为:miR-NC组(空质粒),miR-195-mimics组(模拟序列)。实时荧光定量PCR法检测miR-195表达;MTT检测细胞增殖能力;Transwell侵袭实验检测细胞侵袭力;细胞划痕实验检测细胞转移能力;流式细胞术检测细胞凋亡情况;Western blot检测chemokine 5表达水平;Spearman相关分析miR-195及chemokine 5相关性。荧光素酶实验验证miR-195与chemokine 5的靶向关系。结果:miR-195-mimics组细胞miR-195水平高于miR-NC组(P<0.05);miR-195 mimics组第1、2、3、4 d细胞活力低于miR-NC组(P<0.05);miR-195 mimics组G1细胞高于miR-NC组,G2期、S期细胞低于miR-NC组,G2/S期细胞比值低于miR-NC组(P<0.05);miR-195 mimics组划痕距离大于miR-NC组(P<0.05);miR-195 mimics组细胞侵袭数低于miR-NC组(P<0.05);miR-195-mimics组细chemokine 5蛋白含量低于miR-NC组(P<0.05);miR-195 mRNA水平与chemokine 5蛋白含量负相关(r=-0.398,P=0.00);miR-195可直接靶向chemokine 5。结论:miR-195可通过靶向chemokine 5促进胃癌MGC803及NCI-N87细胞的增殖、转移及侵袭。

Objective:To explore the molecular mechanism of miR-195 targeting chemokine 5 inhibiting proliferation,apoptosis,invasion and metastasis of gastric cancer cells.Methods:Gastric cancer cell lines MGC803,NCI-N87 were selected.According to the transfection,it was divided into:miR-NC group(null plasmid)and miR-195-mimics group(simulated sequence).The expression of miR-195 in cells was detected by real-time fluorescence quantitative PCR.The effect of transfected miR-195 on the proliferation of tumor cells was detected by MTT assay.Cell invasion was analysed by Transwell.Cell migration was detected by wound healing.Cell apoptosis was checked by flow cytometry.The protein levels of chemokine 5 were determined by western blot.The correlation between the expressions of miR-195 and chemokine 5 was analysis by Spearman.Results:The level of miR-195 in miR-195-mimics group was higher than that in miR-NC group(P<0.05);The cell viability of miR-195 group on the 1,2,3 and 4 day was lower than those of miR-NC group(P<0.05);The number of G1 cells in miR-195-mimics group was higher than that in miR-NC group;The number of G2 and S-phase cells was lower than that in miR-NC group;The ratio of G2/S-phase cells was lower than that in miR-NC group(P<0.05);The scratch distance of miR-195 group was greater than that of miR-NC group(P<0.05);The cell invasion number of miR-195 group was lower than that of miR-NC group(P<0.05);The content of chemokine 5 protein in miR-195-mimics group was lower than that in miR-NC group;The level of miR-195 mRNA was negatively correlated with the content of chemokine 5 protein(r=-0.398,P=0.00).miR-195 could directly target chemokine 5 protein.Conclusion:MiR-195 promotes the proliferation,metastasis and invasion of MGC803 and NCI-N87 cells by targeting chemokine 5.