State Key Laboratory of Biotherapy, West China Hospital and West China Medical School, Sichuan University, Ke-yuan Road 4, No. 1 Gao-peng Street, Chengdu, Sichuan, The People's Republic of China .
Cancer Biother Radiopharm. 2011 Aug;26(4):427-36. doi: 10.1089/cbr.2010.0922. Epub 2011 Jul 28.
Polo-like kinase 1 (PLK1) showing a high expression in various kinds of tumors is considered a candidate target for cancer therapy. The aim of our study was to explore the effects of silencing PLK1 gene on human colorectal carcinoma cell line HCT-116 in vitro and in vivo. In vitro, the plasmids generating short hairpin RNA (shRNA)-targeting PLK1 were transfected into HCT-116 by using FugeneHD reagent, and the silencing potency was measured by RT-PCR, western blot, flow cytometry, and Caspase-Glo 3/7 assay, respectively. In vivo, the growth inhibition capacity of PLK1-shRNA on HCT-116 xenograft was measured in nude mice. Then, the silencing effect of PLK1 was analyzed by RT-PCR, western blot, and immunohistochemistry, respectively. Apoptosis, angiogenesis, and proliferation in tumor tissues were measured by TUNEL, CD31, and PCNA stain, respectively. The RNA interference targeting PLK1 significantly decreased the expression of PLK1 in vitro. More importantly, anti-PLK1 treatment in HCT-116 xenograft decreased tumor weight by 81.58% compared with the control group (p<0.001), accompanied with decreased PLK1 mRNA and protein expression, increased cell apoptosis, and reduced angiogenesis and proliferation (p<0.001). Our study showed that knockdown of PLK1 by shRNA might be the potential therapeutic approach against human colorectal carcinoma.
丝氨酸/苏氨酸激酶 polo 样激酶 1(PLK1)在各种肿瘤中呈高表达,被认为是癌症治疗的候选靶点。本研究旨在探讨沉默 PLK1 基因对体外和体内人结直肠癌细胞系 HCT-116 的影响。在体外,使用 FugeneHD 试剂将产生短发夹 RNA(shRNA)靶向 PLK1 的质粒转染至 HCT-116 中,分别通过 RT-PCR、western blot、流式细胞术和 Caspase-Glo 3/7 测定来测量沉默效力。在体内,在裸鼠中测量 PLK1-shRNA 对 HCT-116 异种移植物的生长抑制能力。然后,通过 RT-PCR、western blot 和免疫组织化学分别分析 PLK1 的沉默效果。通过 TUNEL、CD31 和 PCNA 染色分别测量肿瘤组织中的细胞凋亡、血管生成和增殖。靶向 PLK1 的 RNA 干扰显著降低了体外 PLK1 的表达。更重要的是,与对照组相比,抗-PLK1 治疗 HCT-116 异种移植物使肿瘤重量降低了 81.58%(p<0.001),同时降低了 PLK1 mRNA 和蛋白表达,增加了细胞凋亡,并减少了血管生成和增殖(p<0.001)。我们的研究表明,shRNA 敲低 PLK1 可能是治疗人结直肠癌的潜在方法。