Han Zheng, Huang Xiaodong, Liu Meng, Tan Jie, Zhu Qingxi, Wang Yanfen, Cai Yishan, Chen Aifang, Tian Xia
Department of Gastroenterology, Wuhan Third Hospital, Wuhan 430060, China.
Department of Gastroenterology, Wuhan Third Hospital, Wuhan 430060, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 May;33(5):606-610.
Objective To investigate the effects of cadherin 17 (CDH17) on the proliferation and apoptosis of noscapine-resistant human SW480 colon cancer cells. Methods The level of CDH17 in noscapine-resistant human SW480 colon cancer cells was knocked down by small interfering RNA (siRNA), and the silence was confirmed by Western blotting and real-time quantitative PCR. Transfected SW480 cells were treated with noscapine, and then the proliferation and cell viability of SW480 cells were measured by MTT assay and plate clone formation assay, respectively; the apoptosis of SW480 cells was detected by flow cytometry combined with annexinV-FITC/PI staining; the expressions of poly-ADP-ribose polymerase (PARP) and caspase-3 were determined by Western blotting. Results Compared with NC-siRNA group and control group, the expression levels of CDH17 protein and mRNA were down-regulated in the CDH17-siRNA-transfected SW480 cell lines. After noscapine treatment, compared with NC-siRNA and control group, the colony-forming ratio and cell viability were significantly lower in CDH17-siRNA -transfected cell lines, and the expression levels of cleaved-PARP and cleaved- caspase-3 were up-regulated in CDH17-siRNA group, and the cell apoptosis rate increased. Conclusion Knock-down of CDH17 in SW480 cells can effectively inhibit cell proliferation and promote cell apoptosis as well as improve SW480 cell sensitivity to narcotine.
目的 探讨钙黏蛋白17（CDH17）对纳可丁耐药的人SW480结肠癌细胞增殖和凋亡的影响。方法 采用小干扰RNA（siRNA）敲低纳可丁耐药的人SW480结肠癌细胞中CDH17的水平，通过蛋白质免疫印迹法和实时定量PCR法确认沉默效果。对转染后的SW480细胞进行纳可丁处理，分别采用MTT法和平板克隆形成试验检测SW480细胞的增殖和细胞活力；采用流式细胞术结合膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色检测SW480细胞的凋亡情况；通过蛋白质免疫印迹法检测聚ADP核糖聚合酶（PARP）和半胱天冬酶-3的表达。结果 与NC-siRNA组和对照组相比，CDH17-siRNA转染的SW480细胞系中CDH17蛋白和mRNA的表达水平下调。纳可丁处理后，与NC-siRNA组和对照组相比，CDH17-siRNA转染的细胞系克隆形成率和细胞活力显著降低，CDH17-siRNA组中裂解的PARP和裂解的半胱天冬酶-3的表达水平上调，细胞凋亡率增加。结论 敲低SW480细胞中的CDH17可有效抑制细胞增殖，促进细胞凋亡，并提高SW480细胞对纳可丁的敏感性。
