过表达microRNA-7通过下调CGG结合蛋白1的表达抑制人肺癌细胞生长  

Over-expressed microRNA-7 inhibits the growth of human lung cancer cells via suppressing CGGBP1 expression

作　　者：胡燕[1] 廖珍媛[1] 陈超[1] 秦娜琳[1] 郑静[1] 田丹[1] 李永菊[1] 朱顺飞 罗军敏[1] 徐林[1] 

机构地区：[1]贵州省遵义医学院免疫学教研室、贵州省免疫学研究生教育创新基地,贵州遵义563000

出　　处：《细胞与分子免疫学杂志》2014年第2期125-130,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81260398);教育部新世纪优秀人才计划(NCET-12-0661);贵州省国际合作项目(10C315)

摘　　要：目的探讨过表达microRNA-7(miR-7)对人肺癌细胞体内外生长的作用和可能机制。方法利用真核表达载体pcDNA3.1(-)-pri-miR-7(p-miR-7),体外瞬时转染95D人肺癌细胞,MTT法和克隆形成实验检测95D细胞的增殖变化;免疫荧光技术检测95D细胞Ki-67和CGG结合蛋白(CGGBP1)的表达;建立人肺癌裸鼠肿瘤模型,肿瘤局部注射p-miR-7真核表达载体,测量肿瘤大小并观察小鼠生存时间;实时PCR检测肿瘤组织中miR-7表达水平;免疫组织化学技术检测组织中Ki-67和CGGBP1蛋白的表达。结果过表达miR-7可明显抑制肺癌细胞的体外增殖(P<0.05),Ki-67和CGGBP1的表达显著减少(P<0.05);体内实验结果显示,局部注射p-miR-7组中miR-7表达水平明显增加,同时荷瘤裸鼠的肿瘤生长缓慢,生存时间明显延长(P<0.05)。肿瘤组织中Ki-67和CGGBP1蛋白的表达量均显著减少(P<0.05)。结论过表达miR-7可显著抑制人肺癌细胞的体内外生长,可能与其下调肿瘤生长相关蛋白CGGBP1的表达有关。Objective To investigate the effect of microRNA-？ （miR-7） over-expression on the growth of human lung cancer ceils in vivo and in vitro and explore its possible mechanism. Methods The eukaryotic expression vector of pcDNA3.1 encoding miR-？ （p-miR-7） was transiently transfected into human lung cancer 95D cells in vitro. The proliferation of cells was detected by M＇l-r assay and colony formation assay. Moreover, the expressions of nuclear antigen Ki-67 and CGG binding protein t （CGGBP1） were detected by immunofluorescence assay. Human lung cancer model in nude mice was established. Next, p-miR-？ vector was directly injected into local tumor tissue. Then, tumor size was measured and the survival time of mice was observed. The expression level of miR-？ in the tumor tissue was determined by real-time PCR. Finally, the expressions of Ki-67 and CGGBP1 were detected by immunohistochemistry. Results Over-expression of miR-7 could significantly inhibit the growth of 95D cells in vitro （P〈0.05）, accompanied by the remarkably reduced expressions of Ki-67 and CGGBP1 （P 〈 0.05）. Moreover, compared with those in control group, the expression level of miR-7 increased significantly in p-miR-7 injected group （ P 〈 0.05）. Meanwhile, the growth of tumors in injected group was slower than in the control group. Consistently, the survival time of mice was dramatically prolonged in p-miR-7 injected group （P 〈 0.05）. The expression levels of Ki-67 and CGGBP1 also remarkably decreased in tumor tissue （ P 〈 0.05 ）. Conclusion Over-expression of miR-7 could significantly inhibit the growth of human lung cancer cells in vivo and in vitro, which might be related to the down-regulated expression of tumor growth-associated protein CGGBPI.

关 键 词：肺癌 miR-7 人肺癌95D细胞 细胞生长 CGG结合蛋白1 

分 类 号：R734.2[医药卫生—肿瘤] R392-33[医药卫生—临床医学]