靶向结肠癌 SW620细胞 ZEB2-shRN A重组体构建及功能研究

摘要

Objective :To construct zinc‐finger E‐box binding homeobox 2(ZEB2)short hairpin RNA(shRNA) and ob‐serve the grow th characteristics of SW620 colorectal cancer cells after shRNA‐mediated ZEB2 gene silencing .Methods :Interference plasmid shZEB2 slow virus infected cells target to ZEB2 genes was constructed by pLVX‐shRNA2 vector , stably transfected cell lines were screened by G418 ,RT‐PCR and Western blot were used to detect ZEB2 expression in SW620 cells on the levels of Genes and protein respectively .The characteristics of ZEB2‐shRNA transfected SW620 cells were analyzed from the assays of colony formation ,wound healing and tumorigenicity in nude mice ,respectively . Results :ShZEB2 stably transfected cell lines were successfully constructed .The reduction of ZEB2 expression in ZEB2‐shRNA transfected SW620 cells led to the significantly decrease of the abilities colony capability ,migration and tumori‐genicity in nude mice(P<0 .05) .Conclusion:The expression of ZEB2 in colon cancer cell line SW620 can be reduced by RNA interference technique and can decrease their tumorigenicity .These results suggest that the ZEB2 can be an effec‐tive therapeutic target for human colorectal cancer treatment .%目的：利用RNA干扰技术，降低结肠癌SW620细胞中E盒结合锌指蛋白2（ZEB2）基因的表达，观察ZEB2低表达对SW620细胞的生长影响。方法：用pLVX‐shRNA2载体构建针对人ZEB2基因的干扰质粒shZEB2慢病毒感染细胞，G418筛选稳定转染细胞株，通过RT‐PCR和Western blot检测ZEB2在SW620细胞中表达。采用克隆形成检测转染细胞克隆能力，划痕试验检测转染细胞的迁移力，RT‐PCR检测转染细胞ZEB2基因表达水平，致瘤试验检测转染细胞在裸鼠的致瘤性。结果：成功构建 shZEB2稳定转染的细胞株，shZEB2‐SW620内ZEB2表达下降，导致shZEB2‐SW620在细胞克隆能力、迁移力及致瘤性方面均下降（P＜0．05）。结论：用RNA干扰技术可以靶向降低结肠癌细胞株SW620内ZEB2的表达，使其致瘤性降低，提示ZEB2可作为分子靶点用于结肠癌靶向治疗。