微小RNA-221作用蓬乱蛋白2影响前列腺癌细胞系的侵袭功能研究

目的:探讨微小RNA-221(microRNA-221,miR-221)在前列腺癌细胞系中的表达,对神经内分泌样(NE)转化及其侵袭功能的影响.方法:以Northern blot检测雄激素依赖性前列腺癌(LNCaP) 细胞系,雄激素非依赖性前列腺癌(LNCaP-AI) 细胞系中miRNA的表达;细胞转染法检测在雄激素剥夺环境中LNCaP和LNCaP-AI细胞系中miR-221的作用;细胞增殖检测法检测细胞在不同阶段的生长增殖水平;Transwell法检测转染细胞的侵袭能力;定量逆转录聚合酶链反应和Western blot检测转染细胞中神经元特异性烯醇化酶及蓬乱蛋白2 (DVL2)表达的变化.结果:与LNCaP相比,miR-221在 LNCaP-AI中明显高表达.通过转染使miR-221在LNCaP中高表达可促进细胞的神经元特异性烯醇酶的表达,加速NE转化.而在LNCaP-AI中下调miR-221水平可增强靶基因DVL2的表达,并增强LNCaP-AI的迁移和侵袭能力(P<0.01).结论:LNCaP和LNCaP-AI中miR-221表达有差异.miR-221可促进前列腺癌细胞的NE转化,可能是导致前列腺癌雄激素非依赖转化的重要原因.MiR-221可通过作用DVL2调节前列腺癌细胞的转移和侵袭.

MicroRNA-221 expression affects invasion potential of human prostate carcinoma cell lines by regulating disheveled-2

Objective:To evaluate the effect of microRNA-221(miR-221) on the neuroendocrine(NE) differentiation and invasive function of prostate cancer cells.Methods:The expressions of 7 miRNAs in the cell lines of androgen-dependent carcinoma of prostate(LNCaP) and androgen-independent carcinoma of prostate(LNCaP-AI) were detected by Northern blotting.The LNCaP and LNCaP-AI cells cultured in androgen-depleted medium were transfected with different synthetic miR-221, and their invasive abilities were evaluated by a matrigel invasion assay;the cell growth was assessed by using the cell counting kit-8 cell proliferation assay at different time points;the expression of neuron-specific enolase and dishevelled-2 (DVL2)during NE differentiation and migration was respectively measured by quantificational real-time polymerase chain reaction and Western blot.Results:The miR-221 level was increased significantly in LNCaP-AI cell line when compared with LNCaP cell line.Overexpression of miR-221 in LNCaP cells significantly promoted neuron-specific enolase expression and accelerated NE differentiation;suppression of miR-221 expression improved the abilities of migration and invasion of LNCaP-AI cells;meanwhile, the DVL2 mRNA and protein levels were upregulated after the transfection of anti-miR-221 in LNCaP-AI cells(P<0.01).Conclusions:There is a significant difference in miR-221 expression between androgen-dependent prostate cancer and androgen-independent prostate cancer cells.MiR-221 contributes to the NE differentiation of LNCaP cells, which may be the reason of androgen-independence.MiR-221 may regulate the migration of LNCaP-AI cells through DVL2 in advanced prostate cancer.