Department of Urology, General Hospital of Jinan Military Command, 25 Shifan Road, 250031 Jinan, China.
Med Oncol. 2012 Jun;29(2):815-22. doi: 10.1007/s12032-011-9934-8. Epub 2011 Apr 13.
The aim of this study is to evaluate the effect of the variation of miR-221 on the prostate cancer cells' NE differentiation and invasive function and to examine the function of miR-221 in plasma as a blood-based miRNA biomarker candidate for CaP. The expression of 7 miRNAs in LNCaP, LNCaP-AI, and PC3 prostate cancer cell lines was detected by Northern blotting. LNCaP and LNCaP-AI cells cultured in androgen-depleted medium were transfected with different synthetic miRs. The ability of invasiveness was evaluated by a Matrigel invasion assay. Cell growth was assessed by using the CCK-8 cell proliferation assay at different times. The expression of NSE and DVL2 during the neuroendocrine phenotype and migration were measured by qRT-PCR and Western blot. The level of miR-221 in the prostate cancer samples was measured by qRT-PCR. MiR-221 was significantly increased compared AIPC with ADPC cell lines. Overexpression of miR-221 in LNCaP cells significantly increased the level of NSE expression and induced NE differentiation. Knocking down the level of miR-221 expression with antagonist miR-221 in the LNCaP-AI cell line increased migration and invasion (P < 0.01). DVL2 protein level was up-regulated after transfection of anti-miR-221. MiR-221 was up-regulated in CaP plasma (P < 0.01). We demonstrate a significant difference in miR-221 expression between ADPC and AIPC. MiR-221 may contribute to NE differentiation, which may be the cause for AIPC. We also suggest that miR-221 may control the migration of AIPC cells through DVL2, working as a key regulator in advanced CaP. The role of miR-221 in other target mRNA needs to be further investigated.
本研究旨在评估 miR-221 的变异对前列腺癌细胞去势抵抗性神经内分泌分化和侵袭功能的影响，并探讨其作为前列腺癌血液 miRNA 生物标志物候选物在血浆中的功能。通过 Northern blot 检测了 LNCaP、LNCaP-AI 和 PC3 前列腺癌细胞系中 7 种 miRNA 的表达。在雄激素耗竭培养基中培养的 LNCaP 和 LNCaP-AI 细胞用不同合成的 miR 转染。通过 Matrigel 侵袭实验评估侵袭能力。在不同时间使用 CCK-8 细胞增殖实验评估细胞生长。通过 qRT-PCR 和 Western blot 测量神经内分泌表型和迁移过程中 NSE 和 DVL2 的表达。通过 qRT-PCR 测量前列腺癌样本中 miR-221 的水平。与 ADPC 细胞系相比，AIPC 中 miR-221 的表达显著增加。在 LNCaP 细胞中过表达 miR-221 显著增加 NSE 表达水平并诱导去势抵抗性神经内分泌分化。在 LNCaP-AI 细胞系中用拮抗剂 miR-221 下调 miR-221 的表达水平会增加迁移和侵袭（P<0.01）。转染 anti-miR-221 后 DVL2 蛋白水平上调。MiR-221 在 CaP 血浆中上调（P<0.01）。我们证明了 ADPC 和 AIPC 之间 miR-221 表达存在显著差异。MiR-221 可能有助于去势抵抗性神经内分泌分化，这可能是 AIPC 的原因。我们还表明，miR-221 通过 DVL2 控制 AIPC 细胞的迁移，作为晚期 CaP 的关键调节剂。miR-221 在其他靶 mRNA 中的作用需要进一步研究。
