siRNA下调CacyBP/SIP表达对人乳腺癌细胞增殖、凋亡及侵袭的影响  

Effects of down-regulated CacyBP/SIP expression by small interfering RNA on proliferation,apoptosis and invasion of human breast cancer cells

作　　者：王燕[1] 王宁菊[2] 廉斌[2] 丁静[2] 李少林[1] 

机构地区：[1]重庆医科大学放射医学教研室,重庆400016 [2]宁夏医科大学总医院肿瘤医院肿瘤内科,宁夏回族自治区银川750004

出　　处：《肿瘤》2013年第8期670-675,共6页Tumor

基　　金：宁夏回族自治区自然科学基金重点资助项目(编号:NZ10113)

摘　　要：目的:探讨靶向钙周期素结合蛋白(calcyclin binding protein/Siah-1-interacting protein,CacyBP/SIP)基因的小干扰RNA(small interfering,siRNA)对人乳腺癌细胞MDA-MB-231增殖、凋亡及侵袭的影响。方法:化学合成3条针对CacyBP/SIP基因的特异性siRNA,采用脂质体法转染MDA-MB-231细胞;分别采用实时荧光定量-PCR和蛋白质印迹法检测siRNA转染前后MDA-MB-231细胞中CacyBP/SIP mRNA及蛋白的表达情况。分别采用平板克隆形成实验、MTT法、FCM法及Transwell小室实验检测CacyBP/SIP-siRNA对MDA-MB-231细胞增殖、凋亡及侵袭能力的影响。结果:与空白对照组和阴性对照组比较,转染CacyBP/SIP-siRNA后乳腺癌MDA-MB-231细胞中CacyBP/SIP mRNA(P<0.05)及蛋白的表达被下调。MDA-MB-231细胞增殖受到抑制,克隆形成能力下降,凋亡率增加(P<0.05),侵袭能力下降(P<0.05)。结论:CacyBP/SIP-siRNA能下调MDA-MB-231细胞中CacyBP/SIP mRNA和蛋白的表达,从而抑制MDA-MB-231细胞的增殖及侵袭能力,诱导细胞凋亡,因此CacyBP/SIP可能参与了乳腺癌的恶性生物学行为。Objective： To investigate the effects of down-regulated expression of CacyBP/SIP,a calcyclin and Siah-1-interacting protein,induced by siRNA （small interfering RNA） on proliferation,apoptosis and invasion of human breast cancer MDA-MB-231 cells.Methods： Chemically synthesized three siRNAs targeting CacyBP /SIP gene were transfected into MDA-MB-231 cells by LipofectAMINE TM 2000.The expressions of CacyBP/SIP mRNA and protein were detected by RFQ-PCR （real-time fluorogenic quantitative-PCR） and Western blotting,respectively.The MTT method,colony formation assay,FCM （flow cytometry） and Transwell assay were performed to detect the proliferation ability,cell colony formation rate,apoptotic rate and invasion ability of MDA-MB-231 cells,respectively.Results： As compared with the blank control and the negative control groups,the expression levels of CacyBP/SIP mRNA （P〈0.05） and protein in MDA-MB-231 cells were reduced after transfection with CacyBP/SIP-siRNA.The proliferation of MDA-MB-231 cells was inhibited,the ability of colony formation was weakened,and the invasion ability was decreased （P〈0.05）.The apoptosis rate was increased （P〈0.05）.Conclusion： CacyBP/SIP-siRNA can down-regulate the expression levels of CacyBP/SIP mRNA and protein in MDA-MB-231 cells,inhibit the proliferation and invasion abilities of MDA-MB-231 cells,and induce the apoptosis of MDA-MB-231 cells effectively.CacyBP/SIP may be involved in malignant behavior of breast cancer.

关 键 词：乳腺肿瘤 RNA 小分子干扰 细胞增殖 细胞凋亡 基因 CacyBP SIP 

分 类 号：R737.9[医药卫生—肿瘤]