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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 expression in prostate cancer cell lines on neuroendocrine transformation and invasion. Methods Northern blot was used to detect the expression changes of seven microRNAs in LNCaP and LNCaP-AI prostate cancer cell lines. The effects of miR-221 in LNCaP and LNCaP-AI cell lines in androgen-deprivation -8 method was used to detect the growth and proliferation of cells in different stages; Transwell method was used to detect the invasion ability of transfected cells; qRT-PCR and Western blot were used to detect the expression of neuron-specific enolase (NSE) and disheveled-2 DVL2) expression changes. Results miR-221 was significantly overexpressed in LNCaP-AI, an androgen-independent prostate cancer (AIPC) cell line, compared to androgen-dependent prostate cancer (ADPC) cell line LNCaP. The expression of miR-221 in LNCaP cell line by transfection can promote the expression of NSE and accelerate its neuroendocrine-like differentiation. However, the down-regulation of miR-221 in LNCaP-AI cells increased the expression of target gene DVL2 and enhanced the migration and invasion ability of LNCaP-AI cells. Conclusions This experiment confirmed the expression difference of miR-221 in AIPC and ADPC cell lines. miR-221 promotes the neuroendocrine transformation of prostate cancer cells, which may be an important reason for the androgen-independent transformation of prostate cancer. MiR-221 regulates the metastasis and invasion of advanced prostate cancer cells through the action of DVL2.