摘要
目的:研究BRMS1基因对卵巢上皮性癌(卵巢癌)细胞侵袭、转移的作用及可能机制。方法:应用脂质体介导法将BRMS1-siRNA转染人卵巢癌细胞株OVCAR3细胞内(实验组),设转染不干扰任何基因的NC-siRNA的OVCAR3细胞为阴性对照组,未进行任何转染的为空白对照组。BRMS1基因有效沉默后,用Tr-answell侵袭、迁移实验检测OVCAR3细胞侵袭、迁移能力的变化,并采用蛋白质印迹法及免疫细胞化学法检测蛋白NF-κB p65、uPA表达的变化。结果:人卵巢癌细胞OVCAR3中BRMS1基因成功沉默。Tr-answell侵袭实验中,实验组转入底层膜的细胞数〔(190±8.5)个〕明显高于阴性对照组和空白对照组〔分别为(144±7.8)和(146±6.8)个,P<0.05〕。Transwell迁移实验中,实验组转入底层膜的细胞数〔(231±8.9)个〕明显高于阴性对照组和空白对照组〔分别为(177±9.7)和(182±7.9)个,P<0.05〕。在实验组中NF-κBp65、uPA蛋白表达均显著增强。结论:BRMS1基因可抑制卵巢癌细胞的转移,机制可能与NF-κB信号转导通路有关,通过抑制NF-κB的表达,下调uPA的表达,从而实现抑制卵巢癌的远处转移。
OBJECTIVE：To study the effect and mechanism of BRMS1 on the invasion and metastasis of ovarian cancer cell. METHODS： BRMS1 small interfering RNA（BRMSI-siRNA） was transfected into human ovarian cancer cell line OVCAR3 by liposome transfection method. The cells were divided into 3 groups：experimental group with BRMSI-siRNA, negative control group with siRNA that did not impact any gene, blank control group without any transfection. Invasion and migration in cells were performed u- sing transweli invasion and migration assay. Assessment of protein levels of NF-κB p65 and urokinase-type plasminogen activator （uPA） were observed by Western blot and immunocytochemistry. RESULTS： BRMS1 gene was silenced in OVCAR3 cell line successfully. In transwell invasion assay, the numbers of cells in lower chamber passing through the membrane in transfected group, negative control group and blank control group were 190 ±8.5, 144± 7.8 and 146±6.8 respectively,while in transwell migration assay, the numbers of cells in lower chamber passing through the membrane in transfected group, negative control group and blank control group were 231±8.9, 177±9.7 and 182±7.9 respectively, both with significant diferences among 3 groups（P〈0.05）. While the protein expression of NF-κB p65 and uPA were dramatically elevated by BRMS1 gene silenced. CONCLUSION： BRMS1 gene can suppress metastasis of ovarian cancer through reducing the expression of NF-κB, by which down-regulated the expression of uPA, which may be one of the mechanisms of suppress distant metastasis of ovarian cancer.
出处
《中华肿瘤防治杂志》
CAS
2011年第15期1161-1165,共5页
Chinese Journal of Cancer Prevention and Treatment
基金
广东省科技厅项目(2009B060700080)
广州市科技局项目(2010GN-E00221)
关键词
卵巢肿瘤
肿瘤转移
BRMS1基因
ovarian neoplasms
neoplasm metastasis
genes, breast cancer metastasis suppressor 1
