摘要
目的:研究沉默核糖体合成调控因子1(regulator of ribosome synthesis 1,RRS 1)基因表达对乳腺癌BT549细胞增殖、凋亡、迁移和侵袭的影响,并探讨可能的作用机制。方法:实时荧光定量PCR及蛋白质印迹法检测乳腺癌BT549细胞及正常乳腺HMEC细胞中RRS1 mRNA及蛋白的表达量。构建携带有RRS1-shRNA的慢病毒表达载体,并感染BT549细胞。分别采用实时荧光定量PCR和蛋白质印迹法鉴定RRS 1基因的沉默效率。MTT法、FCM法、DAPI染色法以及Transwell小室法分别检测沉默RRS 1基因表达后BT549细胞增殖、凋亡、迁移及侵袭能力的变化,并再次采用蛋白质印迹法检测对BT549细胞中凋亡相关蛋白p53和小鼠双微体2同源物(mouse double minute 2 homolog,MDM2)表达的影响。结果:乳腺癌BT549细胞中RRS1 mRNA及蛋白的表达水平均高于其在正常乳腺HMEC细胞中的表达水平(P值均<0.01)。RRS1-shRNA转入BT549细胞后,RRS1 mRNA和蛋白的表达水平均较对照组明显降低(P值均<0.01)。RRS 1基因沉默后,BT549细胞的增殖活力明显下降(P<0.01),细胞周期被阻滞在G2期(P<0.01),细胞早期凋亡率明显增高(P<0.05),迁移及侵袭能力均明显降低(P值均<0.05),凋亡相关蛋白p53的表达水平明显上调(P<0.05),而MDM2蛋白的表达水平明显下调(P<0.05)。结论:RRS 1基因在乳腺癌细胞中高表达;RRS 1作为一个新发现的乳腺癌相关基因,与乳腺癌细胞的增殖、凋亡、迁移和侵袭功能相关。
Objective： To investigate the effects of silencing the regulatur ul ribosome synthesis 1 （RRS1） gene expression on proliferation, apoptosis, migration and invasion abilities of breast cancer BT549 cells, and to explore the possible mechanism. Methods： The expression levels of RRS1 mRNA and protein in breast cancer BT549 cells and normal mammary gland HMEC cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The lentivirus expression vector carrying RRS1 -shRNA was constructed and transfected into BT549 cells, while the empty vector was used as the control. The silencing efficiency of RRS1 gene was identified by real-time fluorescent quantitative PCR and Western blotting, respectively. Then the proliferation, cell cycle, apoptosis, migration and invasion abilities of BT549 cells transfected with RRSI-shRNA were detected by MTT, FCM, DAPI staining and Transwell chamber assay, respectively. The expression levels of apoptosis-related proteins p53 and mouse double minute 2 homolog （MDM2） in BT549 cells transfected with RRS1 -shRNA were detected by Western blotting. Results： The expression levels of RRS1 mRNA and protein in BT549 cells were significantly higher than those in the normal mammary gland HMEC ceils （both P 〈 0.01）. After transfection with RRSI-shRNA, the expression levels of RRS1 mRNA and protein in BT549 cells were significantly down-regulated as compared with the control group （both P 〈 0.01）. After RRS1 gene silencing, the cell viability was significantly decreased （P 〈 0.01）, the cell cycle was arrested at G2 phase （P 〈 0.01）, the early apoptosis rate was significantly increased （P 〈 0.05）, while the migration and invasion abilities were significantly decreased （both P 〈 0.05）. The expression level of apoptosis-associated p53 protein was significantly up- regulated （P 〈 0.05）, but the expression level of MDM2 protein was significantly down- regulated （P 〈 0.05） in BT549 cells after transfection wit
出处
《肿瘤》
CAS
CSCD
北大核心
2018年第1期35-43,共9页
Tumor
基金
国家自然科学基金面上项目(编号:81472542)~~
关键词
乳腺肿瘤
核糖体合成调控因子1
RNA
小分子干扰
细胞增殖
细胞凋亡
肿瘤转移
Breast neoplasms
Regulator of ribosome synthesis 1
RNA, small interferingCellular proliferation
Apoptosis
Neoplasm metastasis
