Twist1在结直肠癌多药耐药中的作用机制  

Mechanisms of Twist1 role in multidrug resistance of colorectal cancer

作　　者：程家平[1] 文坤明[1] 黎江[2] 曾庆良[1] 陈正权[1] 

机构地区：[1]遵义医学院附属医院胃肠外科,遵义563000 [2]遵义医学院附属医院甲乳外科,遵义563000

出　　处：《临床与实验病理学杂志》2016年第8期860-863,869,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：贵州省科学技术基金[黔科合J字LKZ(2013)50号]

摘　　要：目的探讨Twist1在结直肠癌多药耐药中的作用机制。方法建立人结直肠癌耐奥沙利铂细胞株(SW620/OxR),构建带G418抗性的Twist1-shRNA质粒转染SW620/OxR细胞作为实验组,同时,设置转染带G418抗性阴性对照质粒的SW620/OxR细胞作为阴性对照组,并在转染成功后用G418筛选建立稳定转染细胞株。采用实时荧光定量PCR和Western blot法检测SW620/OxR细胞中Twist1基因和蛋白的表达,采用流式细胞术间接法检测细胞中Twist1、p-STAT3、P-gp、Survivin蛋白变化,Western blot法检测两组细胞Twistl、STAT3、p-STAT3蛋白表达变化。结果 Twist1-shRNA干扰质粒能够明显降低SW620/OxR细胞Twist1 mRNA及蛋白水平。且实验组的E-cadherin表达水平明显降低,vimentin表达水平明显升高。采用流式细胞术检测肿瘤细胞凋亡,阴性对照组细胞凋亡率为5.08%,实验组则明显增高至30.69%。流式细胞术间接法检测实验组中Pgp、Survivin、p-STAT3及Twist1阳性率分别为5.25%、1.93%、2.51%、3.58%,阴性对照组中P-gp、Survivin、p-STAT3及Twist1阳性率分别增加至21.98%、22.36%、19.42%、53.68%,二者差异有统计学意义(P<0.01)。实验组Twist1、STAT3、p-STAT3蛋白相对表达量明显低于阴性对照组,差异有统计学意义(P<0.01)。结论 Twist1可能是结直肠癌多药耐药机制中的关键因子,其可能通过逆向调控STAT3信号通路发挥作用。Purpose To investigate the mechanism of Twistl in the muhidrug resistance of eoloreetal cancer. Methods Oxaliplatin-resistant human eoloreetal cancer cell lines （SW620/OxR） were established, G418 resistance construct transfeeted with the negative control with G418 resistance Twistl-shRNA plasmids were transfeeted into SW620/OxR cells as experimental group, and plasmid SW620/OxR cells were set as negative control group; after a successful transfeetion with G418 screening stably transfected cell lines were established. Using real-time quantitative PCR and Western blot assay Twistl expression was detected in SW620/OxR cell; protein expression of Twistl, p-STAT3, P-gp and survivin was detected by flow cytometry; Western blot assay was used to determine Twistl, STAT3 and p-STAT3 protein expression. Results Twistl-shRNA interference plasmid could significantly reduce levels of mRNA and protein Twistl in SW620/OxR cells. The expression levels of E-eadherin were significantly decreased in the experimental group, while vimentin increased significantly. Flow eytometry showed that apoptotie rate was 5.08% in the negative control group, while apoptotie rate was significantly increased to 30. 69% in the experimental group. Indirect flow eytometry assay showed that the cell percentage of P-gp ＋ , Survivin ＋ and Twistl in the experimental group were 5.25% , 1.93% , 2.51% and 3.58% , while the cell percentage of those protein in the negative control group increased to 21.98% , 22. 36% , 19.42% and 53.68% , respectively, the difference of which was statistieally significant （ P 〈 0. 01 ）. Twistl, STAT3, p-STAT3 protein in the experimental group was significantly lower than that in the negative control group, with statistically significant difference （P 〈 0. 01 ）. Conclusion Twistl probably is a key factor in the mechanism of drug resistance in eoloreetal cancer, which may play a role by reversely regulating STAT3 signaling pathway.

关 键 词：结直肠肿瘤 多药耐药 TWIST1 SW620/OxR 

分 类 号：R735.3[医药卫生—肿瘤]