文章摘要
胡 华,周 忠.miR⁃1247对骨肉瘤细胞增殖、凋亡的影响及机制研究[J].南京医科大学学报,2020,(2):200~204
miR⁃1247对骨肉瘤细胞增殖、凋亡的影响及机制研究
Effects and mechanism of miR⁃1247 on proliferation and apoptosis of human osteosarcoma cells
投稿时间:2018-02-23  
DOI:10.7655/NYDXBNS20200210
中文关键词: miR⁃1247  骨肉瘤  增殖  凋亡
英文关键词: miR⁃1247  osteosarcoma  proliferation  apoptosis
基金项目:
作者单位
胡 华 华中科技大学同济医学院附属武汉中心医院骨科 湖北 武汉 430014 
周 忠 华中科技大学同济医学院附属武汉中心医院骨科 湖北 武汉 430014 
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中文摘要:
      目的:检测microRNA?1247(miR?1247)在骨肉瘤细胞系中的表达及对骨肉瘤细胞增殖和凋亡的影响和机制。方法:采用实时荧光定量PCR(qRT?PCR)测定miR?1247在人正常成骨细胞系(U2OS、Saos?2、143B)及人骨肉瘤细胞系(hFOB1.19)中的表达,将Saos?2细胞系分成两组,即miR?1247过表达组(miR?1247组)和阴性对照组(EV组),采用LipofectamineTM 2000分别转染miR?1247 mimics和阴性对照序列scramble,采用MTT法和流式细胞术测定增殖和凋亡,Western blot检测SOX9和FAM129B的表达。结果:miR?1247在骨肉瘤细胞系中的相对表达量低于人正常成骨细胞系,差异有统计学意义(P < 0.05)。转染后0、1、3 d,EV组与miR?1247组细胞增殖水平差异无统计学意义(P > 0.05);转染后5、7 d,miR?1247组细胞增殖水平低于EV组,差异有统计学意义(P < 0.05)。miR?1247组细胞凋亡率高于EV组,差异有统计学意义(P < 0.01)。 miR?1247组SOX9和FAM129B蛋白表达量低于EV组,差异有统计学意义(P < 0.05)。结论:miR?1247在骨肉瘤细胞系中低表达,miR?1247上调表达可抑制细胞增殖并诱导凋亡,其机制可能与SOX9和FAM129B蛋白下调表达有关。
英文摘要:
      Objective:This study aims to analyze the expression of miR?1247 in human osteosarcoma cell lines,as well as its effect and mechanism on proliferation and apoptosis of osteosarcoma cells. Methods:qRT?PCR was performed to evaluate expression of miR?1247 in human osteosarcoma cell lines(U2OS,Saos?2 and 143B)and human normal osteoblast cell line(hFOB1.19). Saos?2 cell line were divided into two groups,miR?1247 group transfected with miR?1247 mimics and EV group transfected with miR?1247 scramble. MTT assay and flow cytometry was used to assess proliferation and apoptosis ability of cells. Western blot was carried out to determine Sox9 and FAM129B proteins expression. Results:miR?1247 expression level in osteosarcoma cell lines was significantly lower than that in normal osteoblast cell line (P < 0.05). There was no significantly difference in cell proliferation between EV and miR?1247 group after transfect for 0,1,3 d(P > 0.05). The cell proliferation of miR?1247 group was significantly lower than that of EV group after transfect for 5,7 d(P < 0.05). The apoptosis rate of miR?1247 group was significantly higher than that of EV group(P < 0.05). The expression level of SOX9 and FAM129B protein of miR?1247 group was significantly lower than that of EV group(P < 0.05). Conclusion:Osteosarcoma cell lines showed lower miR?1247 expression compare to normal osteoblast cell line. miR?1247 inhibit proliferation and induce apoptosis in osteosarcoma cell lines via down?regulation of Sox9 and FAM129B proteins.
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