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目的:探讨小干扰RNA(small interference RNA,siRNA)介导的细胞色素P4501B1(cytochrome P-4501B1,CYP1B1)基因沉默对二十碳五烯酸(eicosapentaenoic acid,EPA)和花生四烯酸(arachidonic acid,AA)作用下乳腺癌MDA-MB-231细胞增殖的影响。方法:应用细胞计数试剂盒(cell counting kit-8,CCK-8)检测经EPA和AA处理后MDA-MB-231细胞的增殖情况。采用RNA干扰(RNA interference,RNAi)技术干扰CYP1B1的表达,随后应用荧光实时定量PCR(real-time-uorescence quantitative PCR,RFQ-PCR)和蛋白免疫印迹法检测转染效率,以及siRNA干扰后EPA和AA作用下MDA-MB-231细胞中CYP1B1和儿茶酚氧位甲基转移酶(catechol-O-methyltransferase,COMT)mRNA和蛋白的表达情况。采用CCK-8法检测siRNA干扰后EPA和AA作用下MDA-MB-231细胞的增殖情况。结果:EPA处理组的MDA-MB-231细胞数明显少于对照组,而AA处理组的MDA-MB-231细胞则明显多于对照组(P<0.05)。转染CYP1B1siRNA的MDA-MB-231细胞中,CYP1B1mRNA和蛋白的表达均有所下降,而COMT mRNA和蛋白的表达水平则有所上升。CYP1B1siRNA转染的MDA-MB-231细胞的增殖能力下降,且EPA处理组的MDA-MB-231细胞数明显多于阴性对照组(P<0.05)。结论:CYP1B1基因沉默能够抑制MDA-MB-231细胞的增殖,逆转EPA对细胞增殖的抑制作用。EPA可能通过调节CYP1B1的表达来抑制乳腺癌细胞的增殖。
OBJECTIVE: To investigate the effect of silencing cytochrome P-4501B1 (CYP1B1) gene mediated by small interfering RNA (siRNA) on eicosapentaenoic acid (EPA) and arachidonic acid , AA) on the proliferation of breast cancer MDA-MB-231 cells. Methods: Cell counting kit-8 (CCK-8) was used to detect the proliferation of MDA-MB-231 cells treated with EPA and AA. The expression of CYP1B1 was interfered by RNA interference (RNAi) technology. Then the transfection efficiency was detected by real-time-uorescence quantitative PCR (RFQ-PCR) and western blotting, AA on mRNA and protein expression of CYP1B1 and catechol-O-methyltransferase (COMT) in MDA-MB-231 cells. The proliferation of MDA-MB-231 cells under the action of EPA and AA was detected by CCK-8 assay. Results: The number of MDA-MB-231 cells in the EPA-treated group was significantly less than that in the control group, while the MDA-MB-231 cells in the AA-treated group were significantly more than the control group (P <0.05). CYP1B1 siRNA transfected MDA-MB-231 cells, CYP1B1 mRNA and protein expression were decreased, while the COMT mRNA and protein expression levels increased. The proliferation of MDA-MB-231 cells transfected with CYP1B1 siRNA decreased, and the number of MDA-MB-231 cells in the EPA-treated group was significantly more than that in the negative control group (P <0.05). Conclusion: CYP1B1 gene silencing can inhibit the proliferation of MDA-MB-231 cells and reverse the inhibitory effect of EPA on cell proliferation. EPA may inhibit the proliferation of breast cancer cells by regulating the expression of CYP1B1.