Tian Xia, Liu Meng, Zhu Qingxi, Tan Jie, Liu Weijie, Wang Yanfen, Chen Wei, Zou Yanli, Cai Yishan, Han Zheng, Huang Xiaodong
a Department of Gastroenterology , Tongren Hospital of Wuhan University (Wuhan Third Hospital) , Wuhan , China.
Expert Rev Anticancer Ther. 2017 Sep;17(9):857-863. doi: 10.1080/14737140.2017.1344097. Epub 2017 Jun 27.
The aim of the present study was to explore the signaling pathway of noscapine which induces apoptosis by blocking liver-intestine cadherin (CDH17) gene in colon cancer SW480 cells.
Human colon cancer SW480 cells were transfected with CDH17 interference vector and treatment with 10 µmol/L noscapine. The proliferation and apoptosis of SW480 cells were detected by MTT assay and AnnexinV-FITC/PI flow cytometry kit (BD), respectively. Cell invasion were assessed by transwell assays. Apoptosis related proteins (Cyt-c, Bax, Bcl-2 and Bcl-xL) levels were evaluated by western blot.
Compared to the noscapine group, the proliferation was decreased significantly and the apoptosis was increased significantly in SW480 cells of the siCDH17+noscapine group. Cyt-c and Bax protein levels in siCDH17+noscapine group was higher than that of the noscapine group, but Bcl-2 and Bcl-xL protein levels in siCDH17+noscapine group were lower than that of the noscapine group. Moreover, up-expression of CDH17 inhibited the efficacy of noscapine-induced apoptosis in SW480 cells.
We inferred that down-expression of extrinsic CDH17 gene can conspicuously promote apoptosis-inducing effects of noscapine on human colon cancer SW480 cells, which is a novel strategy to improve chemotherapeutic effects on colon cancer.
本研究旨在探讨诺司卡品通过阻断结肠癌SW480细胞中肝肠钙黏蛋白(CDH17)基因诱导细胞凋亡的信号通路。
将CDH17干扰载体转染人结肠癌SW480细胞,并用10 μmol/L诺司卡品处理。分别采用MTT法和AnnexinV-FITC/PI流式细胞术试剂盒(BD)检测SW480细胞的增殖和凋亡情况。通过Transwell实验评估细胞侵袭能力。采用蛋白质免疫印迹法检测凋亡相关蛋白(细胞色素C、Bax、Bcl-2和Bcl-xL)的水平。
与诺司卡品组相比,siCDH17+诺司卡品组SW480细胞的增殖显著降低,凋亡显著增加。siCDH17+诺司卡品组细胞色素C和Bax蛋白水平高于诺司卡品组,但siCDH17+诺司卡品组Bcl-2和Bcl-xL蛋白水平低于诺司卡品组。此外,CDH17的过表达抑制了诺司卡品诱导SW480细胞凋亡的效果。
我们推断,外源性CDH17基因的低表达可显著增强诺司卡品对人结肠癌SW480细胞的凋亡诱导作用,这是提高结肠癌化疗效果的一种新策略。