miR-135a靶向调控STAT6对前列腺癌细胞生物学行为的影响

The effect of miR-135atargeted regulation of STAT6on the biological behavior of prostate cancer cells

目的分析微小RNA-135a(miR-135a)靶向调控信号传导和转录激活因子6(STAT6)对前列腺癌细胞生物学行为的影响。方法选取DU145前列腺癌细胞株,细胞培养后分为空白组、miR-135a阴性对照组、miR-135a转染组,构建miR-135a慢病毒载体,空白组加入常规细胞培养液、生理盐水做空白处理,miR-135a阴性对照组加入miR-135aNC、Lipofectamine 2000试剂行无义序列转染,miR-135a转染组加Hsa-miR-135a、Lipofectamine 2000试剂行细胞转染。鉴定miR-135a转染效率及STAT6mRNA表达量,分析对细胞生物学行为、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶组织抑制剂-2(TIMP-2)、B淋巴细胞瘤-2(Bcl-2)、Bax蛋白的影响。结果与空白组相比,miR-135a阴性对照组miR-135a表达量降低,miR-135a转染组miR-135a表达量升高,差异具有统计学意义(P<0.05),说明miR-135a转染成功。空白组、miR-135a阴性对照组、miR-135a转染组STAT6mRNA表达量、增殖、凋亡率、侵袭、迁移个数、MMP-2、MMP-9、TIMP-2、Bcl-2、Bax蛋白表达比较,差异具有统计学意义(P<0.05);与miR-135a阴性对照组相比,miR-135a转染组STAT6mRNA表达量、细胞增殖率、侵袭、迁移个数、MMP-2、MMP-9、Bcl-2蛋白表达量降低,细胞凋亡率、TIMP-2、Bax蛋白表达量升高,差异具有统计学意义(P<0.05)。结论miR-135a对STAT6表达有一定的调控作用,通过调控MMP-2、MMP-9、TIMP-2表达抑制前列腺癌细胞迁移和侵袭,通过调控Bcl-2、Bax相关蛋白表达抑制前列腺癌细胞增殖,促进其凋亡,为临床前列腺癌靶向治疗提供理论依据。

Objective To analyze the effects of micro RNA-135a(miR-135a)targeting regulatory signal transduction and transcriptional activator 6(STAT6)on the biological behavior of prostate cancer cells.Methods DU145prostate cancer cell line was selected and divided into blank group,miR-135anegative control group and miR-135atransfection group.MiR-135alentiviral vector was constructed.The blank group was treated with conventional cell culture medium and normal saline,while the miR-135anegative control group was added with miR-135aNC and Lipofectamine.The cells in miR-135atransfection group were transfected with hsa-miR-135aand Lipofectamine 2000.To identify miR-135atransfection efficiency and STAT6mRNA expression,and to analyze the effects on cell biological behavior,matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9),matrix metalloproteinase tissue inhibitor-2(TIMP-2),B lymphoma-2(Bcl-2),Bax protein.Results The expression of miR-135ain miR-135anegative control group and miR-135atransfection group was significantly higher than that in blank group(P<0.05),indicating that miR-135a transfection was successful.There were significant differences in STAT6mRNA expression,proliferation,apoptosis rate,invasion,number of migration,MMP-2,MMP-9,TIMP-2,Bcl-2,Bax protein expression in blank group,miR-135a negative control group and transfection group(P<0.05)miR-135aexpression,cell proliferation,invasion,number of migration,MMP-2,MMP-9,Bcl-2protein expression,apoptosis rate and TIMP-2,Bax protein expression were the transfection group were significantly lower than those in the miR-135anegative control group(P<0.05).Conclusion MiR-135acan regulate the expression of STAT6,inhibit the migration and invasion of prostate cancer cells by regulating MMP-2,MMP-9,TIMP-2expression,inhibit the proliferation of prostate cancer cells and promote their apoptosis by regulating the expression of Bcl-2,Bax related proteins.To provide theoretical basis for clinical prostate cancer targe-ted therapy.