miR⁃1247对骨肉瘤细胞增殖、凋亡的影响及机制研究
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Effects and mechanism of miR⁃1247 on proliferation and apoptosis of human osteosarcoma cells
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    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。

    Abstract:

    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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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(自然科学版),2020,(2):200-204

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  • 收稿日期:2018-02-23
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  • 在线发布日期: 2020-03-14
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