Xin T, Zhang F B, Sui G J, Jin X M
Second Affiliated Hospital of Harbin Medical University Heilongjiang 150086, PR China.
Br J Biomed Sci. 2012;69(2):62-6.
The knockdown of Bmi-1 could effectively suppress cancer cell proliferation and tumourigenicity in several cancers. This study aims to investigate whether or not Bmi-1 plays a causative role in the proliferation of ovarian epithelial cancer cells and telomerase activity. The messenger RNA (mRNA) and protein expression levels of Bmi-1 in the human ovarian carcinoma cell line OVCAR-3 were downregulated by Bmi-1 siRNA, as confirmed by real-time polymerase chain reaction (PCR) and Western blot. Cell viability was analysed by MTT assay, and telomerase activity was analysed by a modified telomeric repeat amplification protocol. Targeting Bmi-1 with siRNA inhibited Bmi-1 mRNA over five-fold compared with the control cells, and inhibited Bmi-1 protein expression over three-fold compared with control cells. The viability of the OVCAR-3 ovarian cancer cell line was reduced by Bmi-1 mRNA compared to control cells. Telomerase activity was decreased 22.73% (from 0.33 to 0.255) by Bmi-1 siRNA treatment compared to control cells. As Bmi-1 siRNA depressed telomerase activity, cell immortalisation may be prevented; thus, silencing Bmi-1 may be a potential therapy to manage ovarian cancer.
敲低Bmi-1可有效抑制多种癌症中的癌细胞增殖和致瘤性。本研究旨在探讨Bmi-1是否在卵巢上皮癌细胞增殖和端粒酶活性中起因果作用。通过实时聚合酶链反应（PCR）和蛋白质印迹法证实，Bmi-1小干扰RNA（siRNA）下调了人卵巢癌细胞系OVCAR-3中Bmi-1的信使核糖核酸（mRNA）和蛋白质表达水平。通过MTT法分析细胞活力，通过改良的端粒重复序列扩增法分析端粒酶活性。与对照细胞相比，用siRNA靶向Bmi-1可使Bmi-1 mRNA抑制超过五倍，使Bmi-1蛋白质表达抑制超过三倍。与对照细胞相比，Bmi-1 mRNA降低了OVCAR-３卵巢癌细胞系的活力。与对照细胞相比，Bmi-1 siRNA处理使端粒酶活性降低了22.73％（从0.33降至0.255）。由于Bmi-1 siRNA抑制了端粒酶活性，可能会阻止细胞永生化；因此，沉默Bmi-1可能是治疗卵巢癌的一种潜在疗法。
