Zhang Yuping, Meng Xia, Shi Hongyang, Li Wei, Ming Zongjuan, Zhong Yujie, Deng Wenjing, Zhang Qiuhong, Fan Na, Niu Zequn, Chen Guo'an, Yang Shuanying
Department of Respiratory Diseases, The Second Affiliated Hospital, Medical College, Xi'an Jiaotong University Xi'an 710004, China.
Toracic Surgery Section, Department of Surgery, University of Michigan Ann Arbor, Michigan 48109, USA.
Am J Transl Res. 2016 Apr 15;8(4):1730-7. eCollection 2016.
The aim of this study is to estimate the role of JAK/STAT3 signaling pathway on apoptosis of lung adenocarcinoma induced by icotinib.
EGFR mutation was detected in lung adenocarcinoma cell line PC-9 by ARMS assay; The inhibitory rates of cell proliferation of PC-9 cells which were exposed to different concentrations of icotinib (0100 μMol/L) for different time (2472 h) respectively were evaluated by MTT assay; Apoptosis of PC-9 cells exposed to different concentrations of icotinib (0, 0.1, 1 and 10 μMol/L) for 48 h were evaluated by TUNEL assay; JAK2, STAT3, Bcl-2, Bax mRNA expressions were evaluated by Real-time PCR assay; The protein levels of P-STAT3 and IL-6 were evaluated by Western-blot assay.
Human lung adenocarcinoma cell line PC-9 had an exon 19 deletion mutation in EGFR gene; Followed by treatment of icotinib, the proliferation of PC-9 cells were all inhibited significantly, especially in 48 and 72 h (P<0.01) in all concentrations; The inhibitory rates of cell proliferation in different treating time had statistical significance (P<0.01); Cell apoptosis in different concentrations were increased significantly (P<0.05); Along with the increasing concentrations, gene expression levels of JAK2, STAT3 and Bcl-2 decreased significantly (P<0.05), Bax increased significantly (P<0.05), JAK2/STAT3 ratios increased significantly (P<0.01), and Bcl-2/bax ratios decreased significantly (P<0.01); P-STAT3 and IL-6 protein levels were inhibited significantly in higher concentration.
JAK/STAT3 signaling pathway participates in apoptosis of PC-9 cells induced by icotinib. The most likely mechanism is icotinib inhibited the gene expression levels of JAK2, STAT3 and Bcl-2, so with the P-STAT3 and IL-6 protein levels, and mediated gene Bax overexpression.
本研究旨在评估JAK/STAT3信号通路在埃克替尼诱导肺腺癌细胞凋亡中的作用。
采用ARMS法检测肺腺癌细胞系PC-9中表皮生长因子受体（EGFR）突变情况；采用MTT法检测不同浓度（0100μMol/L）埃克替尼分别作用不同时间（2472小时）后PC-9细胞的增殖抑制率；采用TUNEL法检测不同浓度（0、0.1、1和10μMol/L）埃克替尼作用48小时后PC-9细胞的凋亡情况；采用实时荧光定量PCR法检测JAK2、STAT3、Bcl-2、Bax mRNA表达水平；采用蛋白质免疫印迹法检测磷酸化信号转导子和转录激活子3（P-STAT3）和白细胞介素-6（IL-6）蛋白水平。
人肺腺癌细胞系PC-9的EGFR基因存在19外显子缺失突变；埃克替尼处理后，PC-9细胞的增殖均受到显著抑制，尤其是在各浓度下作用48和72小时时（P<0.01）；不同处理时间的细胞增殖抑制率具有统计学意义（P<
