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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 effect of small interfering (siRNA) targeting the C-reactive protein (CacyBP / SIP) gene on the proliferation and apoptosis of human breast cancer cell line MDA-MB-231 The impact of death and invasion. Methods: Three specific siRNAs targeting CacyBP / SIP gene were chemically synthesized and transfected into MDA-MB-231 cells by lipofectamine 2000. The expression of MDA-MB-231 was detected by real-time fluorescence quantitative PCR and Western blot respectively. 231 cells in CacyBP / SIP mRNA and protein expression. The effects of CacyBP / SIP-siRNA on the proliferation, apoptosis and invasion of MDA-MB-231 cells were detected by plate clone formation assay, MTT assay, FCM assay and Transwell chamber assay. Results: Compared with the blank control group and the negative control group, CacyBP / SIP mRNA (P <0.05) and protein expression in breast cancer MDA-MB-231 cells transfected with CacyBP / SIP-siRNA were down-regulated. The proliferation of MDA-MB-231 cells was inhibited, the ability of colony formation decreased, the apoptosis rate increased (P <0.05), and the invasive ability decreased (P <0.05). CONCLUSION: CacyBP / SIP-siRNA can down-regulate the expression of CacyBP / SIP mRNA and protein in MDA-MB-231 cells, thereby inhibiting the proliferation and invasion of MDA-MB-231 cells and inducing apoptosis. The malignant biological behavior of breast cancer.