miRNA-32-5p对EZH2基因的靶向调控以及对口腔癌细胞CAL-27增殖的影响

摘要/Abstract

摘要： 目的:构建miR-32-5p慢病毒载体,研究其对EZH2基因的靶向调控作用以及对CAL-27细胞增殖作用的影响。方法:分别构建pSicoR-miR-32-5p及pMIR-Report-EZH2过表达载体,应用双荧光素酶报告基因系统、Western 免疫印迹及实时定量荧光PCR方法验证miR-32-5p与EZH2的靶向调控关系,应用MTT法检测过表达miR-32-5p后对CAL-27细胞的增殖调节作用,采用SPSS20.0软件包对数据进行统计学分析。结果:成功构建了pSicoR-miR-32-5p及pMIR-Report-EZH2重组质粒,并证实miR-32-5p过表达可明显抑制EZH2的蛋白及mRNA的表达水平（P<0.05）;抑制miR-32-5p的表达,则明显上调EZH2的蛋白及mRNA的表达水平(P<0.05);MTT结果显示,miR-32-5p可显著抑制CAL-27的增殖。结论:miR-32-5p可通过靶向作用于EZH2-3’UTR的特异序列,直接抑制EZH2基因的表达并调控CAL-27细胞的增殖。

关键词: miRNA- 32-5p, 口腔癌, EZH2, CAL-27

Abstract: PURPOSE: To construct a lentiviral vector of miR- 32-5p and verify its targeted effect on EZH2 gene and its effect on the proliferation of CAL-27 cells. METHODS: pSicoR-miR-32-5p and pMIR-Report-EZH2 over-expression vector were constructed. The targeted effect of miR-32-5p on EZH2 gene was verified by relative luciferase activity, Western blot and real-time PCR test. The effects on CAL-27 cells proliferation of over-expression miR-32-5p were analyzed by MTT assay. SPSS 20.0 software package was used for statistical analysis. RESULTS: The recombinant plasmids of pSicoR-miR- 32-5p and pMIR -Report-EZH2 3'-UTR were constructed successfully. Over-expression of miR-32-5p suppressed the mRNA and protein expression level of EZH2 significantly (P<0.05). Suppression of miR-32-5p expression increased the expression level of EZH2 mRNA and protein significantly(P<0.05).MTT assay showed that the proliferation effect of lentivirus vector of miR-32-5p on CAL-27 cells were significant(P<0.05). CONCLUSIONS: MiR-32-5p can suppress EZH2 gene expression by targeting the specific sequence of EZH2-3'UTR and affect proliferation of human oral carcinoma cell line CAL-27.

Key words: miRNA-32-5p, Oral carcinoma, EZH2, CAL-27

中图分类号:

R392
R78

郭明, 郝文慧, 翟新颖, 侯林妍. miRNA-32-5p对EZH2基因的靶向调控以及对口腔癌细胞CAL-27增殖的影响[J]. 中国口腔颌面外科杂志, 2016, 14(2): 111-116.

GUO Ming, HAO Wen-hui, ZHAI Xin-ying, HOU Lin-yan. miRNA-32-5p target regulating EZH2 and its effect on proliferation of human oral carcinoma cell line CAL-27[J]. China J Oral Maxillofac Surg, 2016, 14(2): 111-116.

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