【摘要】 通过构建hsa-miR-145的重组体pGenesil-1-miR-145,并瞬时转染到人恶性多发性畸胎瘤细胞(NTERA-2)后,利用qRT-PCR法检测hsa-miR-145,以及肿瘤发生相关基因OCT4,SOX2,c-Myc,KLF4,FSCN1在转录水平的表达差异;同时利用MTT法和流式细胞术Annexin V-APC/7-AAD双染法检测hsa-miR-145对NTERA-2细胞增殖的生物学效应和细胞凋亡的影响.结果表明:hsa-miR-145的高表达导致NTERA-2细胞增殖明显减弱,凋亡率增加;转染重组质粒hsa-miR-145后,OCT4,SOX2,c-Myc,FSCN1基因的表达量均有所降低,提示miR-145的过表达可能通过抑制靶基因的表达从而影响生殖肿瘤细胞的生物学功能.更多还原

【Abstract】 hsa-miR-145 sequence was synthesized and cloned into pGenesil-1 to construct recombinant plasmid pGenesil-1-miR-145.Recombinant plasmid was transfect into NTERA-2cell and detected for transcription level by real time PCR.MTT and flow cytometry were used to detected cell proliferation and apoptosis of NTERA-2cells.The expression of OCT4,SOX2,c-Myc,KLF4,FSCN1 were detected by real time PCR.Recombination plasmid pGenesil-1-miR-145 was successfully constructed and miR-145 was detected by real time PCR in NTERA-2 cells transfected with recombinant plasmid.MTT and flow cytometry assay showed that the hsa-miR-145over-expression inhibited cell proliferation and caused cell apoptosis.In NTERA-2 cells,the mRNA level of OCT4,SOX2,c-Myc and FSCN1,were decreased in pGenesil-1-miR-145 mediated group.Altered expression of hsa-miR-145 in the testis cells may play a role in the regulation of target genes associated with cells proliferation and apoptosis.更多还原