【摘要】 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(LNCa P)细胞系,雄激素非依赖性前列腺癌(LNCa P-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LNCa P和LNCa P-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LNCa P相比,miR-221在LNCa P-AI中明显高表达。通过转染使miR-221在LNCa P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LNCa P-AI中下调miR-221水平可增强靶基因DVL2的表达,并增强LNCa P-AI的迁移和侵袭能力(P<0.01)。结论:LNCa P和LNCa P-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。更多还原

【Abstract】 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( LNCa P) and androgen-independent carcinoma of prostate( LNCa P-AI) were detected by Northern blotting. The LNCa P and LNCa P-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 LNCa P-AI cell line when compared with LNCa P cell line. Overexpression of miR-221 in LNCa P cells significantly promoted neuron-specific enolase expression and accelerated NE differentiation; suppression of miR-221 expression improved the abilities of migration and invasion of LNCa P-AI cells; meanwhile,the DVL2 mRNA and protein levels were upregulated after the transfection of anti-miR-221 in LNCa P-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 LNCa P cells,which may be the reason of androgen-independence. MiR-221 may regulate the migration of LNCa P-AI cells through DVL2 in advanced prostate cancer.更多还原