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MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)
Published on: May 16, 2012
MiR-221 expression affects invasion potential of human prostate carcinoma cell lines by targeting DVL2
Chang Zheng1, Sun Yinghao, Jiao Li
1Department of Urology, General Hospital of Jinan Military Command, 25 Shifan Road, 250031 Jinan, China. doctorchangzheng@163.com
Abstract:
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.
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