肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2014年
5期
310-314
,共5页
Survivin%RNA,小分子干扰%药物疗法,联合%细胞凋亡%乳腺肿瘤
Survivin%RNA,小分子榦擾%藥物療法,聯閤%細胞凋亡%乳腺腫瘤
Survivin%RNA,소분자간우%약물요법,연합%세포조망%유선종류
Survivin%RNA,small interfering%Drug therapy,combination%Apoptosis%Breast neoplasms
目的 观察靶向survivin的siRNA联合紫杉醇和表柔比星对乳腺癌MCF-7细胞凋亡及逆转耐药的影响.方法 构建靶向survivin的siRNA表达质粒,转染乳腺癌MCF-7细胞,荧光定量聚合酶链反应(PCR),Western blot技术检测转染前后survivin基因表达的变化,CCK-8法检测靶向survivin的siRNA联合紫杉醇、表柔比星对MCF-7细胞药物敏感性的影响,利用流式细胞术检测细胞凋亡作用.结果 survivin-siRNA-2能有效抑制survivin mRNA(0.30±0.03)和蛋白的表达(0.37±0.09)(P＜0.05);紫杉醇[24、48 h抑制率分别为(38.5±4.0)％、(41.7±6.3)％]、表柔比星[24、48 h抑制率分别为(37.0±8.6)％、(40.6±12.1)％]均可抑制MCF-7细胞增殖并诱导部分细胞凋亡;当与survivin-siRNA联合时上述作用显著加强[紫杉醇24、48 h抑制率分别为(76.0±2.9)％、(85.1±4.0)％;表柔比星24、48 h抑制率分别为(74.6±5.6)％、(82.5±4.8)％](P＜0.05),并且细胞抑制率呈现时间、剂量依赖性.结论 利用RNA干扰阻断survivin基因表达同时联合相应化疗药物可以显著增强MCF-7细胞对药物敏感性并促进其凋亡,该方法在乳腺癌治疗中具有潜在的临床应用价值.
目的 觀察靶嚮survivin的siRNA聯閤紫杉醇和錶柔比星對乳腺癌MCF-7細胞凋亡及逆轉耐藥的影響.方法 構建靶嚮survivin的siRNA錶達質粒,轉染乳腺癌MCF-7細胞,熒光定量聚閤酶鏈反應(PCR),Western blot技術檢測轉染前後survivin基因錶達的變化,CCK-8法檢測靶嚮survivin的siRNA聯閤紫杉醇、錶柔比星對MCF-7細胞藥物敏感性的影響,利用流式細胞術檢測細胞凋亡作用.結果 survivin-siRNA-2能有效抑製survivin mRNA(0.30±0.03)和蛋白的錶達(0.37±0.09)(P＜0.05);紫杉醇[24、48 h抑製率分彆為(38.5±4.0)％、(41.7±6.3)％]、錶柔比星[24、48 h抑製率分彆為(37.0±8.6)％、(40.6±12.1)％]均可抑製MCF-7細胞增殖併誘導部分細胞凋亡;噹與survivin-siRNA聯閤時上述作用顯著加彊[紫杉醇24、48 h抑製率分彆為(76.0±2.9)％、(85.1±4.0)％;錶柔比星24、48 h抑製率分彆為(74.6±5.6)％、(82.5±4.8)％](P＜0.05),併且細胞抑製率呈現時間、劑量依賴性.結論 利用RNA榦擾阻斷survivin基因錶達同時聯閤相應化療藥物可以顯著增彊MCF-7細胞對藥物敏感性併促進其凋亡,該方法在乳腺癌治療中具有潛在的臨床應用價值.
목적 관찰파향survivin적siRNA연합자삼순화표유비성대유선암MCF-7세포조망급역전내약적영향.방법 구건파향survivin적siRNA표체질립,전염유선암MCF-7세포,형광정량취합매련반응(PCR),Western blot기술검측전염전후survivin기인표체적변화,CCK-8법검측파향survivin적siRNA연합자삼순、표유비성대MCF-7세포약물민감성적영향,이용류식세포술검측세포조망작용.결과 survivin-siRNA-2능유효억제survivin mRNA(0.30±0.03)화단백적표체(0.37±0.09)(P＜0.05);자삼순[24、48 h억제솔분별위(38.5±4.0)％、(41.7±6.3)％]、표유비성[24、48 h억제솔분별위(37.0±8.6)％、(40.6±12.1)％]균가억제MCF-7세포증식병유도부분세포조망;당여survivin-siRNA연합시상술작용현저가강[자삼순24、48 h억제솔분별위(76.0±2.9)％、(85.1±4.0)％;표유비성24、48 h억제솔분별위(74.6±5.6)％、(82.5±4.8)％](P＜0.05),병차세포억제솔정현시간、제량의뢰성.결론 이용RNA간우조단survivin기인표체동시연합상응화료약물가이현저증강MCF-7세포대약물민감성병촉진기조망,해방법재유선암치료중구유잠재적림상응용개치.
Objective To investigate the effects of siRNA against survivin combined with the neoadjuvant chemotherapy (paclitaxel and epirubicin) on the apoptosis in breast cancer MCF-7 cell and reverse drug resistance.Methods Molecular cloning technique was applied to construct the expression vector of siRNA against survivin,and effectene transfection reagent was used to transfect MCF-7 cell.Survivin expression was detected by qPCR and Western blot methods.The effects of paclitaxel or epirubicin combined with or without siRNA against survivin,on the drug susceptibility of MCF-7 cell were detected by CCK-8 assay and that of including MCF-7 cell apoptosis were detected by flow cytometry.Results Survivin-siRNA-2 effectively inhibited the expression of survivin mRNA (0.30±0.03) and protein (0.37±0.09) (P ＜ 0.05).Both paclitaxel [IR 24,48 h =(38.5±4.0) ％,(41.7±6.3) ％] and epirubicin [IR 24,48 h =(37.0±8.6) ％,(40.6± 12.1) ％] can suppress the proliferation of MCF-7 cell and induce its apoptosis,and siRNA against survivin combined with chemotherapy drugs enhanced both effects significantly[paclitaxel IR 24,48 h =(76.0±2.9) ％,(85.1±4.0) ％; epirubicin IR 24,48 h =(74.6±5.6) ％,(82.5±4.8) ％] (P ＜ 0.05).The inhibition rate of MCF-7 was concentration and time dependent.Conclusions Blocking survivin gene expression by RNAi,and combining with the appropriate chemotherapy can significantly enhance the sensitivity of MCF-7 cell to drugs and cell apoptosis.This technology has important potential clinical value in the therapy of breast cancer.
