钙黏蛋白17在白细胞介素-10诱导胃癌细胞侵袭中的作用

The roles of cadherin 17 in interleukin - 10 - induced invasion of gastric cancer cells

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.