文章摘要
薛金秋,石 靓,丁 强.RNPC1基因对人乳腺癌细胞SUM1315化疗药物敏感性的影响[J].南京医科大学学报,2019,(5):648~652
RNPC1基因对人乳腺癌细胞SUM1315化疗药物敏感性的影响
Effects of RNPC1 gene chemosensitivity of human breast cancer cells SUM1315
投稿时间:2018-10-11  
DOI:10.7655/NYDXBNS20190504
中文关键词: RNPC1基因  人乳腺癌细胞  化疗敏感性  慢病毒
英文关键词: RNPC1 gene  human breast cancer cells  chemotherapeutic sensitivity  lentivirus
基金项目:国家自然科学基金资助(81572595;81602336)
作者单位
薛金秋 南京医科大学第二附属医院乳腺外科江苏 南京 210011 
石 靓 南京医科大学第一附属医院乳腺外科江苏 南京 210029 
丁 强 南京医科大学第一附属医院乳腺外科江苏 南京 210029 
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中文摘要:
      目的:探讨RNPC1对人乳腺癌细胞SUM1315化疗药物敏感性的影响及相关机制。方法:慢病毒转染技术将转染至SUN1315细胞中,设敲除(shRNPC1)组和过表达RNPC1组、敲除对照(SCR)组和过表达对照(NC)组。采用实时荧光定量PCR和Western blot 方法检测各组SUM1315细胞中RNPC1的mRNA和蛋白水平。用不同浓度的紫杉醇、多西他赛和5?氟尿嘧啶化疗药物处理各组SUM1315细胞,通过CCK?8法测绘细胞存活曲线并计算半数抑制浓度(IC50),紫杉醇、多西他赛和5?氟尿嘧啶处理各组SUM1315细胞72 h,通过流式细胞仪检测细胞凋亡率。结果:敲除RNPC1组RNPC1 mRNA和蛋白相对表达量均低于SCR组,过表达RNPC1组RNPC1 mRNA及蛋白相对表达量均高于NC组(P < 0.05)。紫杉醇、多西他赛和5?氟尿嘧啶处理各组SUM1315细胞72 h后,shRNPC1组IC50明显低于SCR组(P < 0.05),过表达RNPC1组IC50明显高于NC组(P < 0.05)。用紫杉醇、多西他赛和5?氟尿嘧啶处理各组SUM1315细胞72 h后,shRNPC1组凋亡率明显高于SCR组;过表达RNPC1组凋亡率显著低于NC组(P < 0.05)。结论:RNPC1基因能显著降低人乳腺癌细胞SUM1315对紫杉醇、多西他赛和5?氟尿嘧啶化疗药物的敏感性,有望成为乳腺癌治疗的新策略。
英文摘要:
      Objective:To investigate the effects of RNPC1 gene on chemosensitivity of human breast cancer cells SUM1315 and correlated mechanism. Methods:Lentivirus was used to over?express and knock?down RNPC1 in the SUM1315 breast cancer cells. The cells were divided into overexpress RNPC1(RNPC1)group and its control(NC group),knock?down RNPC1(shRNPC1 group)and its control(SCR group). Real?time PCR and Western blot were analyzed the mRNA and protein expression of RNPC1 in SUN1315 cells. The effect of RNPC1 gene on the sensitivity of the chemotherapeuties drug such as paclitaxel,Taxotere and 5?Fluorouracil with different concentration was detected by CCK8. Apoptosis of breast cancer cell after treated with paclitaxel,Taxotere and 5?Fluorouracil were analyzed by flow cytometry. Results:The qRT?PCR and Western blot results showed that the mRNA and protein level of RNPC1 was increased after transfection with RNPC1 overexpression(RNPC1). While,RNPC1 was reduced after transfection with RNPC1 knock?down RNPC1(shRNPC1)lentivirus. After treatment with different concentration of paclitaxel,Taxotere and 5?Fluorouracil,the IC50 of shRNPC1 group was significantly lower than SCR group(P < 0.05). Furthermore,the IC50 of RNPC1 group was significantly higher than NC group(P < 0.05).The flow cytometer assay was used after treatment with paclitaxel,Taxotere and 5?Fluorouracil for 72 h in the experimental groups. The cell apoptosis rate of shRNPC1 group was higher than SCR group,while,the cell apoptosis rate of RNPC1 was lower than NC group. Conclusions:RNPC1 gene can effectively decrease the sensitivity of human breast cancer cells SUM1315 to the chemotherapeutics drug such as paclitaxel,Taxotere and 5?fluorouracil,indicating that RNPC1 gene silencing might be considered as a new therapeutic strategy for breast cancer.
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