miR-98对肝癌细胞增殖和侵袭能力的影响

目的研究microRNA-98(miR-98)对肝癌细胞增殖和侵袭能力的影响及其可能机制。方法将阴性对照、miR-98 mimics、inhibitor阴性对照、miR-98 inhibitor瞬时转入肝癌细胞株,应用噻唑盐(MTT)法、平板集落形成实验、Transwell小室侵袭法、流式细胞周期实验检测miR-98对SMMC7721细胞生长、增殖及侵袭能力的影响。进一步用Western blot检测Cyclin D1的表达水平。结果 MTT实验表明miR-98过表达后,肝癌细胞的生长能力明显高于对照组,平板克隆实验的结果说明miR-98可以使肝癌细胞的克隆形成率由17.33%升高至76.66%、Transwell小室侵袭实验表明miR-98使肝癌细胞的侵袭能力也明显增强。流式细胞周期实验发现miR-98过表达后,肝癌细胞停留在G1=66.71%期和S=35.89%期的数目显著高于对照组G1=38.93%,S=26.62%。而当miR-98被抑制后,SMMC7721肝癌细胞的生长增殖及侵袭能力减弱。Western blot实验检测发现miR-98过表达后,CyclinD1的表达也升高。结论 miR-98可能通过上调Cyclin D1的表达,增强SMMC7721细胞的生长增殖和侵袭能力,提示miR-98可能在肝癌的发生、发展中起重要作用。

Effect of miR-98 on proliferation and invasion of hepatic carcinoma cell line

Objective To explore the impact of miR-98 on proliferation and invasion of hepatocellular carcinoma cell line SMMC772 and its mechanism. Methods Negative control,miR- 98 mimics, inhibitor negative control and miR- 98inhibitor were transfected into SMMC7721 cells,respectively.MTT assay,plate colony formation assay, transwell assay and flow cytometry detection were applied to evaluate the proliferation and invasion ability of SMMC7721 cells which expressed miR-98, miR-98 inhibitor or the control vector. Cyclin D1 expression was inspected with Western blot. Results MTT assay revealed the growth ability of SMMC7732 cell transfected with miR-98 was obviously higher than that of control group.Plat clone experiments showed that miR- 98 was capable of enhancing the cells clone formation rate from 17.33% to 76.66%.Transwell experiments showed that the invasion of SMMC7732 cells were obviously enhanced by miR-98. Flow cytometry detection found that miR- 98 increased cell number of G1 stage and S stage（ G1 = 66.71% and S = 35.89%.） was significantly higher than in control group（G1=38.93%, S = 26.62%）.While the ability of proliferation and invasion decreased after miR-98inhibtion. Western blot experiments found that the protein level of CyclinD1 was increased after transfection with miR- 98.Conclusion miR-98 can upregulate the expression of cyclin D1, and promote proliferation of hepatic carcinoma cell line.These results suggest an important role of miR-98 in the development and progression of hepatic carcinoma.