Wang Xu, Yang Bo, Ma Baojing
The Department of Urology, Tianjin Institute of Urology, The 2nd Hospital of Tianjin Medical University, Tianjin Medical University, Tianjin, 300211, China.
Department of Cardiology, Chinese PLA General Hospital, No. 28 Fu-xing Road, Beijing, 100853, China.
Cancer Chemother Pharmacol. 2016 Nov;78(5):1025-1031. doi: 10.1007/s00280-016-3158-8. Epub 2016 Sep 29.
In this study, we firstly investigated the regulative effect of miR-204 on Sirt1 expression in prostate cancer cells. Then, we examined whether miR-204 downregulation in the cells is a result of UCA1 upregulation. In addition, the regulative effect of UCA1/miR-204/Sirt1 axis on docetaxel sensitivity of prostate cancer cells was studied.
QRT-PCR was performed to detect UCA1, miR-204 and Sirt1 mRNA expression. Western blot assay was performed to assess Sirt1, P-gp and caspase-3 expression. The regulative effect of UCA1/miR-204/Sirt1 axis on docetaxel sensitivity of prostate cancer cells was examined by measurement of docetaxel IC50, dictation of cleaved caspase-3 and flow cytometric analysis of cell apoptosis.
MiR-204 negatively modulated Sirt1 expression in prostate cancer cells. UCA1 upregulation directly resulted in decreased miR-204 expression. UCA1 overexpression resulted in increased Sirt1 expression in PNT2 cells, while knockdown of endogenous UCA1 led to decreased Sirt1 in LNCaP and 22RV1 cells. UCA1 siRNA, Sirt1 siRNA and miR-204 mimics could enhance docetaxel-induced activation of caspase-3 and cell apoptosis in 22RV1/DR cells.
There is a UCA1/miR-204/Sirt1 axis in LNCaP and 22RV1 cells. The UCA1/miR-204/Sirt1 axis plays an important role in modulating in vitro docetaxel sensitivity of the prostate cancer cells.
在本研究中，我们首先研究了miR-204对前列腺癌细胞中Sirt1表达的调节作用。然后，我们检测了细胞中miR-204下调是否是UCA1上调的结果。此外，还研究了UCA1/miR-204/Sirt1轴对前列腺癌细胞多西他赛敏感性的调节作用。
采用qRT-PCR检测UCA1、miR-204和Sirt1 mRNA表达。进行蛋白质免疫印迹分析以评估Sirt1、P-糖蛋白和半胱天冬酶-3的表达。通过测量多西他赛的半数抑制浓度（IC50）、检测裂解的半胱天冬酶-3以及细胞凋亡的流式细胞术分析，研究UCA1/miR-204/Sirt1轴对前列腺癌细胞多西他赛敏感性的调节作用。
miR-204对前列腺癌细胞中Sirt1的表达具有负向调节作用。UCA1上调直接导致miR-204表达降低。UCA1过表达导致PNT2细胞中Sirt1表达增加，而敲低内源性UCA1导致LNCaP和22RV1细胞中Sirt1表达降低。UCA1小干扰RNA（siRNA）、Sirt1 siRNA和miR-204模拟物可增强多西他赛诱导的22RV1/DR细胞中半胱天冬酶-3的激活和细胞凋亡。
LNCaP和22RV1细胞中存在UCA1/miR-204/Sirt1轴。UCA1/miR-204/Sirt1轴在调节前列腺癌细胞体外多西他赛敏感性方面发挥重要作用。
