Dong Honglin, Yao Luyu, Bi Weilin, Wang Fusheng, Song Wei, Lv Yonggang
Department of Vascular Surgery, Second Hospital, Shanxi Medical University, Taiyuan, China.
J Cancer Res Ther. 2015 Oct-Dec;11(4):717-22. doi: 10.4103/0973-1482.147764.
Chemotherapeutic resistance is a main problem in clinical breast cancer therapy. The purpose of our study is to investigate whether the combination of neoadjuvant chemotherapy and survivin siRNA treatment could enhance the therapeutic effect of neoadjuvant chemotherapy using paclitaxel or epirubicin.
The molecular cloning technique was applied to construct the expression vector of siRNA against survivin. Effectene Transfection Reagent was used to transfect plasmids to MCF-7 cells. Survivin expressions were detected by quantitative real-time PCR (qRT-PCR) and Western blot methods. The effect of paclitaxel or epirubicin, with or without the combination of survivin siRNA treatment, on drug susceptibility of MCF-7 cells was detected by CCK-8 assay. MCF-7 cell apoptosis was detected by Flow Cytometry.
Survivin siRNA effectively inhibited the expression of Survivin RNA and protein levels (P < 0.05). Both paclitaxel and epirubicin can suppress the proliferation of MCF-7 cells and induce apoptosis to a certain degree respectively. The combination of survivin siRNA with the two chemotherapy drugs significantly enhanced both effects of the two chemotherapeutics respectively (P < 0.05).
Survivin siRNA combined with the neoadjuvant chemotherapy can significantly enhance the sensitivity of MCF-7 cells to chemotherapeutics and cell apoptosis. This technology has important potential value in the therapeutic study of breast cancer.
化疗耐药是临床乳腺癌治疗中的一个主要问题。本研究的目的是探讨新辅助化疗与生存素小干扰RNA(siRNA)治疗相结合是否能增强使用紫杉醇或表柔比星的新辅助化疗的治疗效果。
应用分子克隆技术构建针对生存素的siRNA表达载体。使用Effectene转染试剂将质粒转染至MCF-7细胞。通过定量实时聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测生存素表达。通过细胞计数试剂盒-8(CCK-8)检测法检测紫杉醇或表柔比星在联合或不联合生存素siRNA治疗情况下对MCF-7细胞药物敏感性的影响。通过流式细胞术检测MCF-7细胞凋亡情况。
生存素siRNA有效抑制了生存素RNA和蛋白质水平的表达(P<0.05)。紫杉醇和表柔比星均可分别在一定程度上抑制MCF-7细胞增殖并诱导凋亡。生存素siRNA与两种化疗药物联合分别显著增强了两种化疗药物的作用效果(P<0.05)。
生存素siRNA与新辅助化疗联合可显著增强MCF-7细胞对化疗药物的敏感性及细胞凋亡。该技术在乳腺癌治疗研究中具有重要的潜在价值。