Department of Urology, Huashan Hospital, Fudan University, Shanghai, China.
Urol Oncol. 2012 Nov-Dec;30(6):920-7. doi: 10.1016/j.urolonc.2010.10.003. Epub 2011 Apr 7.
The human Med19 gene encodes a critical subunit that stabilizes the whole mediator complex. To understand the role of Med19 in bladder cancer, we studied the effects of lentivirus-mediated suppression of Med19 expression on bladder cancer cells in vitro and in vivo.
In this study, immunohistochemical analysis was used to demonstrate the expression of Med19 in human bladder cancer. The lentivirus vectors containing a small hairpin RNA (shRNA) to target Med19 were constructed. After bladder cancer cells (5637 and T24) were infected, RT-PCR and Western blotting were used to measure Med19 expression. The influence of Med19 on the proliferation of bladder cancer cells were assessed using MTT, BrdU, colony formation and tumorigenicity experiments. Cell cycle was analyzed with flow cytometric assay.
Med19 was up-regulated in human bladder cancers compared with adjacent benign tissues by immunohistochemical analysis, but was strongly inhibited in 5637 and T24 bladder cancer cells infected with lentiviruses delivering shRNA against Med19. The down-regulation of Med19 increased the proportion of cells in G0/G1 phases and attenuated the growth of 5637 and T24 cells in vitro. The tumorigenicity of Med19-suppressed T24 cells was decreased after inoculation into nude mice.
Our results suggested that lentiviruses delivering shRNA against Med19 may be a promising tool for bladder cancer therapy.
人类 Med19 基因编码一个关键亚基，可稳定整个中介体复合物。为了了解 Med19 在膀胱癌中的作用，我们研究了慢病毒介导的 Med19 表达抑制对体外和体内膀胱癌细胞的影响。
在这项研究中，免疫组织化学分析用于证明 Med19 在人膀胱癌中的表达。构建了含有靶向 Med19 的短发夹 RNA（shRNA）的慢病毒载体。膀胱癌细胞（5637 和 T24）感染后，采用 RT-PCR 和 Western blot 检测 Med19 表达。采用 MTT、BrdU、集落形成和致瘤性实验评估 Med19 对膀胱癌细胞增殖的影响。采用流式细胞术分析细胞周期。
免疫组织化学分析显示，与相邻良性组织相比，Med19 在人膀胱癌中上调，但在感染针对 Med19 的 shRNA 的慢病毒的 5637 和 T24 膀胱癌细胞中被强烈抑制。下调 Med19 增加了 G0/G1 期细胞的比例，并减弱了 5637 和 T24 细胞在体外的生长。接种 Med19 抑制的 T24 细胞后，其致瘤性降低。
我们的结果表明，针对 Med19 的 shRNA 慢病毒可能是膀胱癌治疗的一种有前途的工具。
