miR-10b-5p靶向JARID2及PTEN/Akt/PI3K信号通路调控人乳腺癌MDA-MB-231细胞增殖和迁移能力

miR-10b-5p targeting JARID2 and PTEN/Akt/PI3K signaling pathway to regulate the proliferation and migration of human breast cancer MDA-MB-231 cells

目的探讨miR-10b-5p靶向Jumonji富含AT结合结构域2(JARID2)及磷脂酶和张力蛋白同源物(PTEN)/蛋白激酶B(Akt)/磷脂酰肌醇3-激酶(PI3K)信号通路调控人乳腺癌M.D.Anderson和MB代表转移性乳腺癌-231(MDA-MB-231)细胞增殖和迁移的影响。方法培养人正常乳腺细胞密歇根癌症基金会-10A(MCF-10A)及人乳腺癌MDA-MB-231细胞,采用Real-time聚合酶链式反应(PCR)法检测两种细胞中miR-10b-5p表达。将MDA-MB-231细胞分为对照组(Lip2000处理后正常培养细胞)、上调拟似物组(Lip2000+miR-10b-5p拟似物mimic转染)、上调转染组(Lip2000+mimic-miR-10b-5p共转染)、下调拟似物组(Lip2000+miR-10b-5p拟似物inhibitor转染)、下调转染组(Lip2000+inhibitor-miR-10b-5p共转染)。采用四甲基偶氮唑盐比色法(MTT)检测各组细胞增殖情况,Transwell检测各组细胞迁移能力,Western blot法检测各组中JARID2及PTEN/Akt/PI3K信号通路蛋白表达。结果MDA-MB-231细胞中miR-10b-5p表达量高于MCF-10A细胞(P<0.05)。上调转染组增殖抑制率低于对照组,下调转染组增殖抑制率高于对照组(P<0.05);上调拟似物组和下调拟似物组增殖抑制率与对照组比较差异无统计学意义。上调转染组细胞数量少于对照组,下调转染组细胞数量多于对照组(P<0.05);上调拟似物组和下调拟似物组与对照组细胞数量比较差异无统计学意义。上调转染组JARID2、PTEN蛋白表达量均高于对照组,PI3K、Akt蛋白表达均低于对照组(P<0.05);下调转染组JARID2、PTEN蛋白表达量均低于对照组,PI3K、Akt蛋白表达均高于对照组(P<0.05)。结论miR-10b-5p可靶向JARID2及PTEN/Akt/PI3K信号通路表达,影响人乳腺癌MDA-MB-231细胞的增殖和迁移能力。

Objective To investigate the miR-10b-5p targeting the Jumonji AT rich interactive domain 2(JARID2)and the phospholipase and tensin homologues(PTEN)/protein kinase B(Akt)/phosphatidylinositol 3-kinase(PI3K)signaling pathway to regulate the proliferation and migration of human breast cancer M.D.Anderson and MB representing metastatic breast cancer-231(MDA-MB-231)cells.Methods Human normal breast cell Michigan Cancer Foundation 10A(MCF-10A)and human breast cancer MDA-MB-231 cells were cultured,and the expression of miR-10b-5p in the two cells was detected by Real-time polymerase chain reaction(PCR).MDA-MB-231 cells were divided into the control group(normal cultured cells after Lip2000 treatment),the upregulated mimetic group(Lip2000+miR-10b-5p mimic transfection),the upregulated transfection group(Lip2000+mimic miR-10b-5p co transfection),the downregulated mimetic group(Lip2000+miR-10b-5p mimic inhibitor transfection),and the downregulated transfection group(Lip2000+inhibitor miR-10b-5p co transfection).The proliferation status of each group was detected by methyl thiazolyl tetrazolium(MTT)assay using tetramethylazolamide as a trace enzyme,Transwell test the cell migration ability of each group,Western blot method was used to detect the expression of JARID2 and PTEN/Akt/PI3K signaling pathway proteins in each group.Results The expression of miR-10b-5p in MDA-MB-231 cells was higher than that in MCF-10A cells(P<0.05).The proliferation inhibition rate in the upregulated transfection group was lower than the control group,and the downregulated transfection group was higher than the control group(P<0.05);there was no significant difference in proliferation inhibition rate between the upregulated mimetic group and downregulated mimetic group compared with the control group.The number of cells in upregulated transfection group was less than the control group,and the number of cells in downegulated transfection group was more than the control group(P<0.05);there was no significant difference in the number of cells between the upregulated mimetic group and the downregulated mimetic group compared with the control group.The expressions of JARID2 and PTEN protein in the upregulated transfection group were higher than the control group,while the expressions of PI3K and Akt protein were lower than the control group(P<0.05);the protein expressions of JARID2 and PTEN in the downregulated transfection group were lower than the control group,while the protein expressions of PI3K and Akt in downregulated transfection group were higher than the control group(P<0.05).Conclusion miR-10b-5p can target the expression of JARID2 and PTEN/Akt/PI3K signaling pathway,and affect the proliferation and migration of human breast cancer MDA-MB-231 cell.