摘要
目的探讨钙黏蛋白17(CDH17)在白细胞介素(IL)-10诱导的胃癌细胞侵袭中的作用及机制。
方法应用Transewell侵袭实验,观察IL-10对胃癌细胞MKN-45侵袭的诱导作用;并通过构建CDH17小干扰RNA(siRNA),沉默胃癌细胞中CDH17蛋白,观察比较IL-10在CDH17沉默前后,对MKN-45细胞侵袭诱导作用的差异,同时采用Western blot检测其中细胞信号通路的活化,并给予相应信号通路抑制剂,观察抑制信号通路活化后细胞侵袭能力的变化。
结果IL-10(100 ng/ml)显著诱导胃癌细胞MKN-45细胞表达CDH17及诱导侵袭(10 250±1 500比2 750±450,P=0.019),同时Western blot检测显示,其磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号明显活化(P=0.041);siRNA转染成功抑制CDH17表达后,IL-10对MKN-45的侵袭诱导作用明显下降(10 250±1 500比2 325±300,P=0.027),同时PI3K/Akt信号活化受抑制。给予上述未转染MKN-45细胞PI3K/Akt信号通路抑制剂Ly294002(10 nmol/L)预孵育后,PI3K/Akt信号活化抑制,IL-10对其的侵袭诱导能力显著下降(2 050±400比11 500±1 750,P=0.016)。
结论IL-10通过CDH17活化胃癌细胞PI3K/Akt信号通路促进其侵袭转移。
Objective To explore the roles and mechanisms of Cadherin 17 (CDH17) in interleu- kin - 10 - induced invasion of gastric cancer cells. Methods We detected the invasion of MKN -45 gas- tric cancer cells induced by interleukin - 10 through Transwell assay. CDH17 small interfering RNA (siRNA) was constructed and transfected in MKN-45 cells to inhibit CDH17 expression. We then com- pared the difference in cell invasion with or without CDH17 expression and detected the activation of corre- sponding signaling pathway by immunoblotting in response to interleukin - 10. We additionally detected inva- sion of MKN -45 cell after inactivation of signaling by using its specific inhibitor. Results Western blotting and transwell assay revealed that interleukin - 10 ( 100 ng/ml) significantly induced expression of CDH17 in MKN - 45 cells ( P = 0. 021 ) and promoted its invasion ( 10 250±1 500 vs. 2 750 ±450, P = 0. 019 ) as well as activated phosphoinositide -3 - kinase (PI3K)/protein kinase B (Akt) signaling pathway (P = 0. 041 ). After successful inhibition of CDH17 expression by siRNA, the interleukin - 10 - indueeed inva- sion of MKN -45 cells was significantly reduced ( 10 250± 1 500 vs. 2 325± 300, P = 0. 027 ) and the activation of PI3K/Akt signaling pathway was also suppressed. Furthermore, preincubation of Ly294002 ( 10 nmol/L), the specific inhibitor of PI3K/Akt signaling pathway, not only prohibit the activation of P13K/Akt signaling pathway but also obviously diminished interleukin - 10 -induced invasion of MKN -45 cells (2 050 ± 400 vs. 11 500 ± 1 750, P = 0. 016 ). Conclusion Interleukin - 10 - promoted invasion of MKN-45 gastric cancer cells is mediated by CDH17 through P13K/Akt signaling pathway.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第2期199-201,共3页
Chinese Journal of Experimental Surgery
基金
2015年武汉市卫生计划生育委员会资助项目(WX15C31)