miRNA-146a真核表达载体的构建及其对肿瘤细胞恶性增殖的影响  

Construction of miRNA-146a expression vector and its effect on tumor cell malignant phenotype

作　　者：罗婷婷[1] 陈彦[2] 文阳安[3] 

机构地区：[1]重庆市妇幼保健院检验科,400013 [2]西南大学医院,重庆400715 [3]重庆医科大学附属第一医院检验科,400016

出　　处：《国际检验医学杂志》2011年第9期934-935,共2页International Journal of Laboratory Medicine

摘　　要：目的构建miRNA-146a真核表达质粒,探讨其在宫颈癌HeLa细胞株中的表达及对HeLa细胞恶性表型的影响。方法人工合成miRNA-146a基因序列,将miRNA-146a基因克隆到的真核表达载体pGeneSil-1中,构建成重组质粒pGeneSil-miR-146a。将miRNA-146a重组表达载体瞬时转染宫颈癌HeLa细胞株,通过荧光定量聚合酶链反应法(PCR)检测miRNA-146a在转录水平的表达。通过MTT法检测miRNA-146a对细胞增殖的影响,通过流式细胞术检测细胞周期的变化。结果经酶切和测序鉴定,证明重组质粒pGeneSil-miR-146a构建成功。重组质粒转染HeLa细胞后,荧光定量PCR证明能有效表达miRNA-146a。MTT检测结果显示转染miRNA-146a后的HeLa细胞活细胞数目明显增多。流式细胞术细胞周期检测结果表明转染miRNA-146a后的HeLa细胞S期细胞数明显增多。结论成功构建了miRNA-146a基因真核表达载体pGeneSil-miR-146a,转染宫颈癌HeLa细胞株后能有效表达miRNA-146a,miRNA-146a基因表达可以促进HeLa细胞的增殖。Objective To construct microRNA-146a （miR 146a）expression vector and explore the effect of miR-146a on the ma lignant phenotype of cervical cancer HeLa cell line. Methods miR 146a sequence was synthesized and cloned into pGeneSil-1 to construct recombinant plasmid pGeneSil miR-146a. Recombinant plasmid was transfect into HeLa cell and detected for transcription level by real time PCR. MTT and flow cytometry were used to detect cell proliferation and cell cycle of HeLa cells. Results Recom-binant plasmid pGeneSil-miR-146a was successfully constructed and miR 146a was detected By real-time PCR in HeLa cells transfected with recombinant plasmid. MTT and flow cytometry assay showed that miR-146a has proliferative effect on HeLa cells. Conclusion miR-146a expression vector was constructed and the expression of miR 146a in HeLa cell could promote cell proliferation.

关 键 词：宫颈肿瘤 细胞增殖 MIR-146A 

分 类 号：R737.33[医药卫生—肿瘤]