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hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究

周静 吴杲 马福家

周静, 吴杲, 马福家. hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究[J]. 药学实践与服务, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
引用本文: 周静, 吴杲, 马福家. hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究[J]. 药学实践与服务, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
Citation: ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009

hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究

doi: 10.3969/j.issn.1006-0111.2019.05.009

Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p

  • 摘要: 目的 研究hsa-miRNA-30a-5p促进肺癌细胞A549增殖的调控机制。 方法 收集临床肺癌标本及癌旁组织5对,用荧光定量法(real time-PCR)和蛋白质印迹法(Western blotting)分别检测肿瘤及其癌旁组织中hsa-miRNA-30a-5p和SCARA5的蛋白含量;生物信息学预测hsa-miRNA-30a-5p与SCARA5基因3'UTR区结合位点,并通过荧光素酶报告基因法进行结合位点验证;构建SCARA5基因沉默表达载体pshRNA-SCARA5,脂质体瞬时转染pshRNA-SCARA5及hsa-miRNA-30a-5p阻遏物(miRNA-30a-5p inhibitor)到A549细胞,转染后48 h,real time-PCR检测细胞内hsa-miRNA-30a-5p含量,Western blotting检测细胞中SCARA5蛋白含量;MTT法检测转染后A549细胞对数期增殖活性。 结果 肺癌组织中SCARA5蛋白表达量明显低于癌旁组织,组间差异显著(P<0.05);肺癌组织中hsa-miRNA-30a-5p含量则明显高于癌旁组织,组间差异显著(P<0.05)。荧光素酶实验显示,与报告基因表达载体单独转染组比较,miRNA-30a拟似物(miRNA-30a-5p mimics)和miRNA-30a-5p inhibitor与野生型荧光素酶报告基因共转染组可以明显抑制或增强荧光素酶活性(P<0.05);miRNA-30a-5p inhibitor转染细胞后48 h,与转染对照组比较,细胞内hsa-miRNA-30a-5p含量明显降低(P<0.05),阴性对照(miRNA-30a-5p NC)转染组和转染对照组细胞内hsa-miRNA-30a-5p含量无显著差异(P>0.05)。转染后24~72 h,miRNA-30a-5p inhibitor转染组A549细胞增值活性明显降低(P<0.05),而pshRNA-SCARA5转染能够逆转miRNA-30a-5p inhibitor增强A549细胞增殖活性的趋势,共转染组与miRNA-30a-5p inhibitor单独转染组及转染对照组比较,差异有统计学意义(P<0.05)。 结论 Hsa-miRNA-30a-5p通过抑制SCARA5基因表达,增强A549细胞的增殖活性,转染miRNA-30a-5p inhibitor,可以抑制A549细胞增殖活性。
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  • [1] 修建平, 杨朝爱, 刘禧澳, 潘乾禹, 韦广旭, 王卫星.  全反式维甲酸对肝星状细胞活化及氧化应激的作用和机制探索 . 药学实践与服务, 2024, 42(7): 1-6. doi: 10.12206/j.issn.2097-2024.202312054
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  • 收稿日期:  2018-12-11
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hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究

doi: 10.3969/j.issn.1006-0111.2019.05.009

摘要: 目的 研究hsa-miRNA-30a-5p促进肺癌细胞A549增殖的调控机制。 方法 收集临床肺癌标本及癌旁组织5对,用荧光定量法(real time-PCR)和蛋白质印迹法(Western blotting)分别检测肿瘤及其癌旁组织中hsa-miRNA-30a-5p和SCARA5的蛋白含量;生物信息学预测hsa-miRNA-30a-5p与SCARA5基因3'UTR区结合位点,并通过荧光素酶报告基因法进行结合位点验证;构建SCARA5基因沉默表达载体pshRNA-SCARA5,脂质体瞬时转染pshRNA-SCARA5及hsa-miRNA-30a-5p阻遏物(miRNA-30a-5p inhibitor)到A549细胞,转染后48 h,real time-PCR检测细胞内hsa-miRNA-30a-5p含量,Western blotting检测细胞中SCARA5蛋白含量;MTT法检测转染后A549细胞对数期增殖活性。 结果 肺癌组织中SCARA5蛋白表达量明显低于癌旁组织,组间差异显著(P<0.05);肺癌组织中hsa-miRNA-30a-5p含量则明显高于癌旁组织,组间差异显著(P<0.05)。荧光素酶实验显示,与报告基因表达载体单独转染组比较,miRNA-30a拟似物(miRNA-30a-5p mimics)和miRNA-30a-5p inhibitor与野生型荧光素酶报告基因共转染组可以明显抑制或增强荧光素酶活性(P<0.05);miRNA-30a-5p inhibitor转染细胞后48 h,与转染对照组比较,细胞内hsa-miRNA-30a-5p含量明显降低(P<0.05),阴性对照(miRNA-30a-5p NC)转染组和转染对照组细胞内hsa-miRNA-30a-5p含量无显著差异(P>0.05)。转染后24~72 h,miRNA-30a-5p inhibitor转染组A549细胞增值活性明显降低(P<0.05),而pshRNA-SCARA5转染能够逆转miRNA-30a-5p inhibitor增强A549细胞增殖活性的趋势,共转染组与miRNA-30a-5p inhibitor单独转染组及转染对照组比较,差异有统计学意义(P<0.05)。 结论 Hsa-miRNA-30a-5p通过抑制SCARA5基因表达,增强A549细胞的增殖活性,转染miRNA-30a-5p inhibitor,可以抑制A549细胞增殖活性。

English Abstract

周静, 吴杲, 马福家. hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究[J]. 药学实践与服务, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
引用本文: 周静, 吴杲, 马福家. hsa-miRNA-30a-5p促进肺癌细胞A549增殖的机制研究[J]. 药学实践与服务, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
Citation: ZHOU Jing, WU Gao, MA Fujia. Study on the mechanism of proliferation of human lung cancer A549 cells regulated by hsa-miRNA-30a-5p[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 433-439. doi: 10.3969/j.issn.1006-0111.2019.05.009
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