Department of Urological Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing, People's Republic of China.
Urology. 2011 Aug;78(2):474.e7-11. doi: 10.1016/j.urology.2011.03.014. Epub 2011 Jun 25.
To investigate the role of phospholipase Cε (PLCε) by silencing PLCε with short hairpin RNA (shRNA) in human bladder cancer cells BIU-87 in vitro and in vivo.
A PLCε shRNA expression vector was transfected into BIU-87 cells, and the expression of PLCε protein was detected by Western blotting. Cell proliferation was determined using the MTT assay, and the cell cycle was detected using flow cytometry. A tumor xenograft experiment was established to evaluate the tumor growth under the condition of PLCε knockdown, and the expression of PLCε, proliferating cell nuclear antigen, and cyclin D1 were detected by Western blotting or immunohistrochemistry.
PLCε shRNA reduced the protein level of PLCε, leading to marked proliferation inhibition and significant cell cycle arrest. Furthermore, PLCε shRNA reduced the tumor xenograft growth implanted with BIU-87 cells. The protein expression of PLCε, proliferating cell nuclear antigen, and cyclin D1 were downregulated in the bladder tumor xenograft.
The knockdown of PLCε by shRNA could inhibit bladder tumor growth and might be an alternative approach for human bladder cancer therapy.
通过短发夹 RNA（shRNA）沉默人膀胱癌 BIU-87 细胞中的磷脂酶 Cε（PLCε），探讨 PLCε 的作用。
将 PLCε shRNA 表达载体转染至 BIU-87 细胞中，采用 Western blot 检测 PLCε 蛋白的表达。采用 MTT 法检测细胞增殖，流式细胞术检测细胞周期。建立肿瘤异种移植实验，在 PLCε 敲低条件下评估肿瘤生长情况，采用 Western blot 或免疫组化法检测 PLCε、增殖细胞核抗原和细胞周期蛋白 D1 的表达。
PLCε shRNA 降低了 PLCε 蛋白水平，导致明显的增殖抑制和显著的细胞周期阻滞。此外，PLCε shRNA 降低了 BIU-87 细胞植入的肿瘤异种移植的生长。在膀胱癌肿瘤异种移植中，PLCε、增殖细胞核抗原和细胞周期蛋白 D1 的蛋白表达下调。
shRNA 沉默 PLCε 可抑制膀胱癌的生长，可能是治疗人类膀胱癌的一种替代方法。
