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    谢安心, 王思源, 张洁, 张惠娟, 唐海欧, 谭敦勇. miR-99b-5p通过靶向TRIB1基因调控乳腺癌MCF-7细胞增殖和凋亡的研究[J]. 蚌埠医科大学学报, 2022, 47(10): 1331-1335. DOI: 10.13898/j.cnki.issn.1000-2200.2022.10.001
    引用本文: 谢安心, 王思源, 张洁, 张惠娟, 唐海欧, 谭敦勇. miR-99b-5p通过靶向TRIB1基因调控乳腺癌MCF-7细胞增殖和凋亡的研究[J]. 蚌埠医科大学学报, 2022, 47(10): 1331-1335. DOI: 10.13898/j.cnki.issn.1000-2200.2022.10.001
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    XIE An-xin, WANG Si-yuan, ZHANG Jie, ZHANG Hui-juan, TANG Hai-ou, TAN Dun-yong. miR-99b-5p regulates the proliferation and apoptosis of breast cancer MCF-7 cells by targeting TRIB1 gene[J]. Journal of Bengbu Medical University, 2022, 47(10): 1331-1335. DOI: 10.13898/j.cnki.issn.1000-2200.2022.10.001
    Citation: XIE An-xin, WANG Si-yuan, ZHANG Jie, ZHANG Hui-juan, TANG Hai-ou, TAN Dun-yong. miR-99b-5p regulates the proliferation and apoptosis of breast cancer MCF-7 cells by targeting TRIB1 gene[J]. Journal of Bengbu Medical University, 2022, 47(10): 1331-1335. DOI: 10.13898/j.cnki.issn.1000-2200.2022.10.001
    shu

    miR-99b-5p通过靶向TRIB1基因调控乳腺癌MCF-7细胞增殖和凋亡的研究

    miR-99b-5p regulates the proliferation and apoptosis of breast cancer MCF-7 cells by targeting TRIB1 gene

      摘要
    • 摘要:
      目的探讨miR-99b-5p靶向Tribbles同源蛋白1(Tribbles pseudokinase 1，TRIB1)基因调控人乳腺癌MCF-7细胞增殖和凋亡。
      方法采用RT-PCR检测人乳腺上皮细胞HBL-100和乳腺癌MCF-7细胞中miR-99b-5p的表达水平，采用Western blotting检测人乳腺上皮细胞HBL-100和乳腺癌MCF-7细胞中TRIB1蛋白表达量。在MCF-7细胞中转染miR-99b-5p mimics或mimics对照，分别记为miR-99b-5p组和miR-NC组，RT-PCR检测细胞中miR-99b-5p表达变化，Western blotting检测TRIB1蛋白表达量，MTT法检测细胞增殖能力，AnnexinV-FITC/PI双染色法检测细胞凋亡水平，双荧光素酶报告基因实验检测miR-99b-5p和TRIB1的靶向关系。
      结果与人乳腺上皮细胞HBL-100比较，乳腺癌MCF-7细胞中miR-99b-5p的表达量明显降低(P < 0.01)，而TRIB1蛋白的表达明显升高(P < 0.01)。与miR-NC组比较，miR-99b-5p组MCF-7细胞中miR-99b-5p的表达量升高(P < 0.05)，TRIB1蛋白表达水平降低(P < 0.05)，细胞增殖能力减弱(P < 0.05)，细胞凋亡率升高(P < 0.05)。双荧光素酶报告基因检测实验结果显示，miR-99b-5p能够明显降低TRIB1-Wt MCF-7细胞相对荧光素酶活性(P < 0.01)，而对TRIB1-Mut MCF-7细胞相对荧光素酶活性无明显影响(P>0.05)。
      结论miR-99b-5p在乳腺癌MCF-7细胞中呈低表达，TRIB1蛋白呈高表达，过表达miR-99b-5p能够靶向抑制TRIB1阻碍MCF-7细胞增殖并促进细胞凋亡。

       

      Abstract:
      ObjectiveTo investigate the regulation of proliferation and apoptosis of human breast cancer MCF-7 cells by miR-99b-5p targeting Tribbles pseudokinase 1(TRIB1) gene.
      MethodsThe expression of miR-99b-5p in human breast epithelial cells HBL-100 and breast cancer MCF-7 cells was detected by RT-PCR, and the expression of TRIB1 protein in human breast epithelial cells HBL-100 and breast cancer MCF-7 cells was detected by Western blotting.After transfection with miR-99b-5p mimics or mimics control in MCF-7 cells, which was named as miR-99b-5p group and miR-NC group, respectively, the expression of miR-99b-5p in the cells was detected by RT-PCR, the expression of TRIB1 protein was detected by Western blotting, the cell proliferation was detected by MTT assay, the apoptosis level was detected by AnnexinV-FITC/PI double staining, and the targeting relationship between miR-99b-5p and TRIB1 was detected by dual-luciferase reporter gene assay.
      ResultsCompared with human breast epithelial cells HBL-100, the expression of miR-99b-5p in breast cancer MCF-7 cells was significantly decreased(P < 0.01), while the expression of TRIB1 protein was significantly increased(P < 0.01).Compared with miR-NC group, the expression of miR-99b-5p in MCF-7 cells in miR-99b-5p group increased(P < 0.05), the expression level of TRIB1 protein decreased(P < 0.05), the cell proliferation ability decreased(P < 0.05), and the apoptosis rate increased(P < 0.05).The results of dual-luciferase reporter gene assay showed that miR-99b-5p significantly reduced the relative luciferase activity in TRIB1-Wt MCF-7 cells(P < 0.01), but had no significant effect on the relative luciferase activity in TRIB1-Mut MCF-7 cells(P>0.05).
      ConclusionsMiR-99b-5p is down-regulated in breast cancer MCF-7 cells, and TRIB1 protein is up-regulated.Overexpression of miR-99b-5p can inhibit the proliferation and promote apoptosis in MCF-7 cells by targeted inhibiting TRIB1.

       

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