在子宫内膜癌ECC-1中他莫西芬通过mir-585调节PAX2蛋白表达

目的研究在子宫内膜癌ECC-1细胞中他莫西芬（tamoxifen，TAM）对PAX2（pairedbox2）蛋白表达的调节作用，寻找在这-过程中起调节作用的microRNA。方法用他莫西芬刺激ECC-1细胞，Western印迹检测PAX2蛋白表达的变化。利用MicrocosmTargets（miRBaseSequencedatabase）预测了PAX2相关的microRNA。用实时定量的方法检测他莫西芬刺激后ECC-1中PAX2相关microRNA表达的变化，找出差异变化明显的microRNA，合成这些microRNA的mimics，转染人ECC-1细胞中，用Western印迹检测其对PAX2蛋白表达的影响。结果他莫西芬刺激ECC-1细胞系后，Western印迹显示PAX2蛋白表达水平较对照组中等程度上调。实时定量PCR结果显示他莫西芬刺激ECC-1细胞后mir-135b＊，mir-604，mir-585，mir-181c＊表达较对照组明显下调。合成mir-135b＊，mir-604，mir-585，mir-181c＊的mimics并转染人ECC-1细胞后，Western印迹结果显示转入mir-585mimics的ECC-1细胞中PAX2蛋白表达较对照组下调。结论他莫西芬刺激可以引起ECC—1细胞中PAX2蛋白表达水平中等程度上调，通过抑制mir-585的表达减少其对PAX2mRNA翻译的抑制可能是这-调节作用中的部分机制。

Tamoxifen Regulates PAX2 Expression via mir-585 in Endometrial Cancer ECC-1 Cells

Objective To find out if tamoxifen regulates PAX2 expression in endometrial cancer ECC-1 cells, and if any microRNA involved in this process. Methods We first stimulated ECC-1 cells with tamoxifen, then used Western Blot to detect any change in PAX2 expression. We used Microcosm Targets （miRBase Sequence database） to predict PAX2 related microRNAs. After tamoxifen stimulation, any change of PAX2 related microRNAs was detected by real-time quantitative PCR . Any significantly changed microRNA was picked out for mimics synthesis, which was later on tranfected into ECC-1 cells . Western Blot was used to detect how these microRNA mimics had affected PAX2 expression. Results Western Blot confirmed a moderate up regulation of PAX2 expression in tamoxifen stimulated ECC-1 cells . Real-time quantitative PCR showed mir-135b ＊ , mir-604, mir-585 and mir-181c ＊ expression were significantly down regulated in tamoxifen stimulated ECC-1 cells . Synthesized mimics of mir-135b ＊ , mir-604, mir-585 and mir-181c ＊ were transfected into ECC-1 cells . Western Blot detected down regulation of PAX2 expression in ECC-1 cells tranfected with mir-585 mimics. Conclusion Tamoxifen could moderately up regulate expression of PAX2 in ECC-1 cells, and inhibition of mir-585 expression may contribute to this tion. regula