摘要
目的评价miR-221在前列腺癌细胞系中表达的变化对其神经内分泌样转化及其侵袭功能的影响。方法以Northern blot检测LNCaP,LNCaP-AI两种前列腺癌细胞系中7种microRNA的表达变化;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;CCK-8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;qRT-PCR和Western blot检测转染的细胞中神经元特异性烯醇化酶(NSE)及dishevelled-2(DVL2)表达的变化。结果与雄激素依赖性前列腺癌(ADPC)的细胞系LNCaP相比,miR-221在雄激素非依赖性前列腺癌(AIPC)的细胞系LNCaP-AI中明显高表达。通过转染使miR-221在LNCaP细胞系中高表达可促进细胞的NSE表达,加速其神经内分泌样分化。而在LNCaP-AI细胞系中下调miR-221水平则会升高靶基因DVL2的表达水平,并增强LNCaP-AI细胞的迁移和侵袭能力。结论该实验证实在AIPC和ADPC细胞系中miR-221存在表达差异。miR-221可促进前列腺癌细胞的神经内分泌样转化,这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调节晚期前列腺癌细胞的转移和侵袭。
Objective To evaluate the effect of miR-221 on the neuroendocrine differentiation and invasive function of prostate cancer cells as a miRNA biomarker candidate for prostate cancer.Methods The expressions of 7 miRNAs in LNCaP,LNCaP-AI prostate cancer cell lines were detected by Northern blotting.LNCaP and LNCaP-AI cells cultured in androgen-depleted medium were transfected with different synthetic miRNAs.Their invasive abilities were evaluated by a matrigel invasion assay.Cell growth was assessed by using the CCK-8 cell proliferation assay at different time points.The expression of neuron-specific enolase and dishevelled-2 during the neuroendocrine differentiation and migration respectively was measured by qRT-PCR and Western blot.Results The miR-221 level was significantly increased in androgen-independent prostate cancer cell line LNCaP-AI when compared with androgen-dependent prostate cancer cell line LNCaP.Overexpression of miR-221 in LNCaP cells significantly increased NSE expression and induced their NE differentiation.The suppression of miR-221 expression with anti-miR-221 increased the abilities of migration and invasion in LNCaP-AI cells.Meanwhile,the DVL2 mRNA and protein levels were upregulated after the transfection of anti-miR-221 in LNCaP-AI cells.Conclusion There was a significant difference in miR-221 expression between ADPC and AIPC cells.MiR-221 contributed to NE differentiation of ADPC cells,which may be the reason of androgen-independence.MiR-221 may regulate the migration of AIPC cells through DVL2 as a key regulator in advanced prostate cancer.
出处
《中华全科医学》
2012年第7期1003-1005,共3页
Chinese Journal of General Practice
