RNAi沉默Sox2表达对结直肠癌细胞5-氟尿嘧啶的增敏作用  

The sensitization effect of RNAi silenced Sox2 expression on 5-FU in colorectal cancer cells

作　　者：员亚晶 徐利晓 孙斌[1] 冯星[1] YUN Ya-jing;XU Li-Xiao;SUN Bin;FENG Xing(Department of Neonatology,Children′s Hospital of Soochow University,Suzhou Jiangsu 215000,China;Institute of Pediatrics,Children′s Hospital of Soochow University,Suzhou Jiangsu 215000,China)

机构地区：[1]苏州大学附属儿童医院新生儿科,江苏苏州215000 [2]苏州大学附属儿童医院儿科研究所,江苏苏州215000

出　　处：《江苏大学学报（医学版）》2020年第1期7-11,17,共6页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81671532);江苏省医学重点学科资助项目(ZDXKA2016013);苏州市小儿内科医学中心(Szzx201504)

摘　　要：目的:研究Sox2与5-氟尿嘧啶(5-FU)诱导人结肠癌细胞(SW620)化疗敏感性及耐药家族基因表达的关系,探讨结直肠癌细胞化疗耐受的机制。方法:经慢病毒包装干扰质粒Sox2沉默SW620细胞后,采用蛋白质印迹法、实时荧光定量PCR和免疫荧光技术检测干扰前后细胞内Sox2的表达变化,CCK8法检测细胞对5-FU敏感性的变化,流式细胞仪检测细胞凋亡率,并检测ABC耐药家族基因的表达情况。结果:靶向Sox2干扰后,Sox2 mRNA及蛋白表达下调,Sox2的阳性表达率显著减少;干扰Sox2后SW620细胞对5-FU敏感性高于对照组细胞,IC 50值明显下降,细胞之间差异有统计学意义(P<0.05);Sox2干扰后细胞凋亡率明显高于对照组,ABC耐药家族基因的表达水平明显下降。结论:靶向干扰Sox2后有效地抑制了Sox2在SW620细胞中的表达,Sox2干扰后显著提高了SW620细胞对5-FU的敏感性,降低了ABC耐药基因的表达,为寻求新的基因靶向治疗提供了可能。Objective:To investigate the relationship between Sox2 and 5-fluorouracil(5-FU)-induced chemotherapy sensitivity of human colon cancer cells(SW620)and gene expression in drug-resistant families,and to explore the mechanism of chemotherapy resistance in colorectal cancer cells.Methods:After silencing SW620 cells with lentiviral packaging interference plasmid Sox2,Western blot,RT-PCR and immunofluorescence staining were used to detect the changes in the expression of Sox2 in cells before and after interference,CCK8 assay was used to detect the sensitivity of cells to 5-FU,Flow cytometry was used to detect the apoptosis rate of cells and the expression of drug-resistant family genes.Results:After targeting Sox2 interference,the expression of Sox2 mRNA and protein was down-regulated,and the positive expression rate of Sox2 was significantly reduced.After interfering with Sox2,the sensitivity of SW620 cells to 5-FU was higher than that of control cells,the IC50 value decreased significantly,and there was a significant statistical difference between cells(P<0.05).The apoptosis rate of cells after Sox2 interference was significantly higher than that of control cells,and the expression level of ABC drug-resistant family genes decreased significantly after Sox2 interference.Conclusion:After interfering with Sox2,the expression of Sox2 in SW620 cells was effectively inhibited.After interfering with Sox2,the sensitivity of SW620 cells to 5-FU was significantly improved,and the expression level of ABC drug-resistant genes was decreased,providing the possibility for seeking new gene targeted therapy.

关 键 词：SOX2 结直肠癌 5-氟尿嘧啶 耐药基因 化疗敏感 

分 类 号：R735.35[医药卫生—肿瘤]