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作 者:张红鸽[1] 范秉琳[1] 嵇晓辉[1] 蔡新华[1] 朱武凌[1]
机构地区:[1]新乡医学院基础医学院病理科,河南新乡453003
出 处:《中国肿瘤生物治疗杂志》2013年第1期48-51,共4页Chinese Journal of Cancer Biotherapy
基 金:河南省杰出青年科学基金资助项目(No.0512000800)~~
摘 要:目的:探讨microRNA-100(miR-100)对人肝癌HepG2细胞中polo样激酶1(polo-like kinase 1,Plk1)表达和细胞凋亡的影响。方法:通过oligofectamine介导,将miR-100 mimics转染入HepG2细胞中,RT-PCR和细胞免疫荧光法检测Plk1 mRNA和蛋白的表达,并采用AnnexinⅤ-FITC试剂盒检测HepG2细胞的凋亡情况。结果:miR-100 mimics成功转染HepG2细胞,转染效率为(88.75±2.22)%。转染48 h后,miR-100 mimics组HepG2细胞Plk1 mRNA的表达水平明显低于阴性对照组、空白对照组和脂质体组[(0.71±0.01)vs(0.95±0.01)、(0.92±0.02)、(0.93±0.02),均P<0.01];转染72 h后,miR-100 mimics组Plk1蛋白几乎不表达,同时其细胞凋亡率明显高于阴性对照、空白对照组和脂质体组[(26.95±6.72)%vs(15.03±5.12)%、(6.88±3.71)%、(9.00±3.37)%,均P<0.05]。结论:miR-100能够抑制Plk1基因的表达,从而促进肝癌HepG2细胞的凋亡。Objective:To explore the effects of microRNA-100(miR-100)on expression of polo-like kinase 1 (Plk1) and the apoptosis of human hepatic carcinoma HepG2 cells. Methods: HepG2 cells were transfected with miR-100 mimics by oligofectamine. RT-PCR and immunofluorescence were used to analyze Plk1 mRNA and protein expressions in HepG2 cells, respectively. Moreover, the apoptosis of HepG2 cells was detected by AnnexinⅤ-FITC kit. Results: HepG2 cells were successfully transfected by miR-100 mimics and the transfection efficiency was (88.75±2.22) %. 48 h after transfection, the expression of Plk1 mRNA decreased significantly in the miR-100 mimics transfection group compared with the negative control, blank control, and liposome groups (/[0.71±0.01/] vs /[0.95±0.01/], /[0.92±0.02/], /[0.93±002/],P〈0.01). 72 h after transfection, Plk1 protein expression was almost undetectable in HepG2 cells transfected with miR-100 mimics. Meanwhile, the cell apoptosis rate in the miR-100 mimics group was significantly increased in comparison with those in the negative control, blank control, and liposome groups (/[26.95±6.72/]% vs /[15.03±512/]%, /[6.88±3.71/]%, /[9.00±3.37/]%, P〈0.05). Conclusion: miR-100 can inhibit the expression of Plk1 gene, therefore promoting the apoptosis of hepatic carcinoma HepG2 cells.
关 键 词:肝癌 HEPG2细胞 microRNA-100 POLO样激酶1 凋亡
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