RNA干扰靶向抑制胰岛素样生长因子2mRNA结合蛋白1基因对肾癌细胞的抑制作用及其机制

Inhibitory effect of RNA interference targeting insulin - like growth factor 2 mRNA - binding protein 1 gene on renal cell carcinoma and its mechanism

目的探讨RNA干扰（RNAi）抑制胰岛素样生长因子2mRNA结合蛋白1（IGF2BPl）基因对肾癌细胞活力和凋亡的影响及其机制。方法以人胚肾HEK293细胞为对照组细胞，通过Westernblot检测人肾癌786-0、A498、ACHN和CaKi-2细胞中IGF2BPl的蛋白表达。将设计并合成的IGF2BP1小干扰RNA（siRNA）经LipofectamineTM2000转染CaKi-2细胞，Westernblot检测转染后的细胞中IGF2BP1、Wnt／β-连环蛋白（B-catenin）信号通路关键分子p-catenin及增殖凋亡相关靶基因增殖细胞核抗原（PCNA）、生存素（Survivin）的蛋白表达。结果786-（0．415±0．038）、A498（0．278±0．030）、ACHN（0．326±0．032）和CaKi-2（0．557±0．053）细胞中IGF2BPl的表达均明显高于在HEK293细胞（0．062±0．009）表达（P=0．003），选择CaKi-2细胞为研究对象。IGF2BPlsiRNA转染后的细胞中IGF2BPI（0．073±0．010）、B-catenin（0．178±0．021）、PCNA（0．147±0．016）、Survivin（0．095±0．010）的蛋白表达均明显低于NC组（0．485±0．055）、（0．547±0．050）、（0．246±0．028）、（0．168±0．018），细胞活力（0．557,c0．045）明显低于NC组（0．804±0．057，细胞凋亡率（21．48±1．54）％明显高于NC组（2．46±0．27）％，差异均有统计学意义（P=0．000）。结论IGF2BPl在肾癌细胞表达升高，抑制其表达后可明显降低细胞活力和诱导凋亡，机制可能是Wnt／β-catenin信号通路的降低。

Objective To investigate the effect of insulin -like growth factor 2 mRNA -binding protein 1 （IGF2BP1） gene by RNA interference （RNAi） on the activity and apoptosis of renal cell carci- noma. Methods The human embryonic kidney HEK293 cells were used as the control group, and the ex- pression of IGF2BP1 in human renal carcinoma 786 -O, A498, ACHN and CaKi -2 cells was detected by Western blotting. The IGF2BP1 small interfering RNA （siRNA） was designed and synthesized, and trans- fected into CaKi - 2 cells by LipofectamineTM 2000. The expression of IGF2BP1, ± - eatenin, proliferating cell nuclear antigen （PCNA） and Survivin proteins was detected by Western blotting. Results The expres- sion of IGF2BP1 in786-0 （0.415 e0.038）, A498 （0.278 e0.030）, ACHN （0.326 ±0.032） and CaKi-2 （0.557 ± 0.053） cells was significantly higher than that in HEK293 cells （0.062 ± 0. 009） （P =0. 003）, and CaKi -2 cells were selected as the research objects. The expression of IGF2BP1 （0.073±0.010）, catenin （0.178 ±0.021）, PCNA （0.147 ±0.016） and Survivin （0.095 ±0.010） in the ceils transfected with IGF2BP1 siRNA decreased obviously as compared with NC group [（0.485 ± 0.055）, （0.547 ±0.050）, （0.246 ±0.028） and （0. 168 ±0.018）±, the cell viability （ 0. 557 ± 0. 045 ） decreased obviously as compared with NC group （0.804 ± 0. 057 ） , and the apoptosisrate [ （21.48 ± 1.54） % ] increased obviously as compared with NC group [ （2. 46 ± 0. 27） % ], with the difference being statistically significant （ P = 0. 000）. Conclusion The expression of IGF2BP1 in renal cell carcinoma cells is increased, and the inhibition of IGF2BP1 expression can significantly reduce cell vi- ability and induce apoptosis, probably by down- regulating the Wnt/β -catenin signaling pathway.