MiR-503靶向bcl-2增强BEL-7402细胞对顺铂的敏感性
发布时间：2019-02-14 17:26
【摘要】：目的:探讨miR-503是否能通过调控bcl-2的表达增强BEL-7402细胞对顺铂的敏感性。方法:采用实时荧光定量PCR检测肝癌细胞中miR-503和bcl-2 mRNA的表达水平;Western印迹观察肝癌细胞中Bcl-2蛋白的表达水平;脂质体转染法将miR-503模拟物瞬时转染至BEL-7402细胞;生物信息学软件预测miR-503潜在靶基因;双荧光素酶活性验证miR-503潜在的靶位点;MTT法检测细胞药物敏感性的改变。结果:相比于HL-7702细胞,miR-503在BEL-7402细胞中表达水平下调,Bcl-2蛋白的表达水平上调;miR-503能够与bcl-2靶向结合,下调bcl-2的表达;miR-503转染组的细胞活力较miRNA阴性对照组显著降低。结论:MiR-503可能通过靶向bcl-2增强BEL-7402细胞对顺铂的药物敏感性,抑制肝癌细胞增殖。
[Abstract]:Aim: to investigate whether miR-503 can enhance the sensitivity of BEL-7402 cells to cisplatin by regulating the expression of bcl-2. Methods: the expression of miR-503 and bcl-2 mRNA in hepatoma cells was detected by real-time fluorescence quantitative PCR, and the expression of Bcl-2 protein was detected by Western blot. MiR-503 mimics were transiently transfected into BEL-7402 cells by liposome transfection; bioinformatics software was used to predict the potential target genes of miR-503; double luciferase activity was used to verify the potential target sites of miR-503; and the changes in drug sensitivity of cells were detected by MTT assay. Results: compared with HL-7702 cells, the expression of miR-503 in BEL-7402 cells was down-regulated, the expression of Bcl-2 protein was up-regulated, and miR-503 could target bcl-2 and down-regulate the expression of bcl-2. The cell viability of miR-503 transfection group was significantly lower than that of miRNA negative control group. Conclusion: MiR-503 may enhance the chemosensitivity of BEL-7402 cells to cisplatin by targeting bcl-2 and inhibit the proliferation of HCC cells.
【作者单位】： 南华大学生物研究所;南华大学湖南省分子靶标新药研究协同创新中心;南华大学药物药理研究所;南华大学肿瘤研究所;
【基金】：国家自然科学基金(81372579) 湖南省教育厅科学研究项目(17C1402,14C0998) 湖南省研究生创新项目(CX2015B386)~~
【分类号】：R735.7
本文编号：2422434
[Abstract]:Aim: to investigate whether miR-503 can enhance the sensitivity of BEL-7402 cells to cisplatin by regulating the expression of bcl-2. Methods: the expression of miR-503 and bcl-2 mRNA in hepatoma cells was detected by real-time fluorescence quantitative PCR, and the expression of Bcl-2 protein was detected by Western blot. MiR-503 mimics were transiently transfected into BEL-7402 cells by liposome transfection; bioinformatics software was used to predict the potential target genes of miR-503; double luciferase activity was used to verify the potential target sites of miR-503; and the changes in drug sensitivity of cells were detected by MTT assay. Results: compared with HL-7702 cells, the expression of miR-503 in BEL-7402 cells was down-regulated, the expression of Bcl-2 protein was up-regulated, and miR-503 could target bcl-2 and down-regulate the expression of bcl-2. The cell viability of miR-503 transfection group was significantly lower than that of miRNA negative control group. Conclusion: MiR-503 may enhance the chemosensitivity of BEL-7402 cells to cisplatin by targeting bcl-2 and inhibit the proliferation of HCC cells.
【作者单位】： 南华大学生物研究所;南华大学湖南省分子靶标新药研究协同创新中心;南华大学药物药理研究所;南华大学肿瘤研究所;
【基金】：国家自然科学基金(81372579) 湖南省教育厅科学研究项目(17C1402,14C0998) 湖南省研究生创新项目(CX2015B386)~~
【分类号】：R735.7
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