地塞米松抑制人卵巢癌细胞HO-8910增殖的分子机制:RhoB信号通路的作用  被引量:13

Mechanism of inhibiting proliferation of human ovarian cancer cells of the line HO-8910 by dexamethasone:the role of RhoB signaling pathway

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作  者:陈玉霞[1] 李宗斌[1] 刁飞[1] 王燕[1] 卢建[1] 

机构地区:[1]第二军医大学病理生理学教研室,上海200433

出  处:《中华医学杂志》2006年第20期1400-1404,共5页National Medical Journal of China

基  金:国家自然科学基金资助项目(30200100)

摘  要:目的观察地塞米松(Dex)对人卵巢癌细胞HO-8910增殖的影响,探讨小G蛋白RhoB信号通路在其中的作用。方法四甲基偶氮唑盐比色法(MTT法)和软琼脂实验检测细胞的增殖;RT-PCR法检测RhoB mRNA的表达;Western印迹检测RhoB及其上下游信号分子磷酸化Akt(p-Akt)、p21cip1/waf1和p27蛋白质的表达;报告基因技术检测p21cip1/waf1基因的转录。结果Dex可以明显抑制HO-8910细胞锚依赖性和非依赖性方式的增殖,100nM Dex处理细胞6d,抑制率为31%。100nM Dex处理细胞可以明显上调RhoB mRNA和蛋白质的表达(为对照的2.5倍);同时伴有p-Akt蛋白质表达的降低(24h时约为对照的50%)、p27蛋白质表达的增加(36h时比对照增加了3.3倍)以及p21cip1/waf1转录和蛋白质表达增多(24h时约为对照的1.7倍)。RhoB过表达可以降低细胞的生长速度,并能增强Dex的增殖抑制作用(100nMDex处理6d,抑制率为44%);而RhoB表达阻断后可逆转Dex的这一效应(抑制率约为13%)。结论Dex抑制HO-8910细胞增殖的机制可能与抑制PI3K/p-Akt信号,从而上调RhoB蛋白质,使RhoB下游信号分子p21cip1/waf1和p27蛋白质表达增加有关。Objective To observe the effect of dexamethasone (Dex) on the proliferation of human ovarian cancer cells of the line HO-8910, and explore the role of RhoB signaling pathway in this process. Methods Human ovarian cancer cells of the line HO-8910he were cultured in culture fluids with or without different concentrations of Dex. The cell growth levels in anchor-dependent and anchor-independent manner were detected by MTT and soft agar assay. Another H)-8910 cells were inoculated in gel with different concentrations of Dex. HO-8910 was transfected with the eukaryotic expression plasmid RhoB-wt, blank plasmids pcDNA3 and RhoB-RNAi, and then the mRNA expression of RhoB, a small GTPase gene, was examined by semi-quantitative RT-PCR. and the protein expressions of RhoB, p-Akt, and p21^cip1/waf1 and p27, both cychn kinase inhibitors (CDIs), were detected by Western blotting. HO-8910 cells were cotransfected with the reporter gene p21-1uc containing p21 promoter and marker reporter gene pRL-tk-luc, then treated with Dex for 24 h. Western blotting was used to detect the transcription of p21^cip1/waf1 gene. Results The RhoB mRNA expression was significantly increased 2 hours after the treatment of 100 nM Dex, and peaked 4 hours later as high as 2.5 times that of the control group. Western blotting showed that the RhoB protein expression increased along the increase of the Des concentration. The protein expression of RhoB in the HO-8910 cells transfected with RhoB-wt was 2.02 times that in the HO-8910 cells transfected with blank plasmid, and the protein expression of RhoB in the HO-8910 cells transfected with RhoB-RNAi was 36% of that of the blank plasmid group ( P 〈 0.01 ). The HO-8910 cell proliferation of the RhoB-RNA1 group was not significantly different from that of the control group, however, the proliferation of the HO-8910 cell treated by 100 nM Dex for 6 days was significantly inhibited with an inhibifionr rate of 13% ( P 〈 0. 01 ). Western blotting showed that Dex down-regulated the p-Akt pr

关 键 词:地塞米松 RHOB GTP结合蛋白质 磷酸化酶激酶 原癌基因蛋白质p21(ras) 基因产物 rex 

分 类 号:R737.31[医药卫生—肿瘤] R739.5[医药卫生—临床医学]

 

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