Shaozhang Zhou,Zhixin Dong,Jinyi Lv,Aiping Zeng,Huilin Wang,Ruiling Ning,Xiangqun Song. The role of the HGF/c-Met signaling pathway in crizotinib-induced apoptosis in lung cancer with c-Met amplification. Oncol Transl Med, 2017, 3: 116-126.
The role of the HGF/c-Met signaling pathway in crizotinib-induced apoptosis in lung cancer with c-Met amplification
Received:December 04, 2016  Revised:June 13, 2017
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KeyWord:HGF/c-MET signaling pathway; H1993 cells; H2228 cells; crizotinib; apoptosis
Author NameAffiliationE-mail
Shaozhang Zhou The Second Department of Chemotherapy, The Affiliated Tumor Hospital, Guangxi Medical University, Guangxi 530021, China zhoushaozhang@qq.com 
Zhixin Dong Guangxi Medical University Graduate School, Guangxi 530021, China  
Jinyi Lv Guangxi Medical University Graduate School, Guangxi 530021, China  
Aiping Zeng The Second Department of Chemotherapy, The Affiliated Tumor Hospital, Guangxi Medical University, Guangxi 530021, China  
Huilin Wang The Second Department of Chemotherapy, The Affiliated Tumor Hospital, Guangxi Medical University, Guangxi 530021, China  
Ruiling Ning The Second Department of Chemotherapy, The Affiliated Tumor Hospital, Guangxi Medical University, Guangxi 530021, China  
Xiangqun Song Affiliated Tumor Hospital of Guangxi Medical University xiangquns@163.com 
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Abstract:
      Objective?This study aimed to study the role of the HGF/c-Met signaling pathway in crizotinib-induced apoptosis of various lung adenocarcinoma cell lines and xenograft tumor models. Methods?In vitro, H2228, H1993, and A549 cells were treated with crizotinib. The inhibition of proliferation was quantitated by a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was quantified by flow cytometry. Expression of key proteins of the HGF/c-Met signaling pathway was examined by western blotting. In vivo, H1993 and A549 tumor cell xenograft models were established. Immunohistochemical analysis was used to determine protein expression of HGF and c-MET and the amount of phospho-c-MET (p-c-Met). Real-time quantitative polymerase chain reaction (PCR) was applied to examine the messenger RNA (mRNA) expression of c-MET and serine/ threonine protein kinase (AKT). The expression and activation of the key proteins were evaluated by western blotting. Results?In vitro, the growth of H1993, H2228, and A549 cells was inhibited after crizotinib treatment for 72 h. Apoptotic rates of H1993 and H2228 cells increased with the crizotinib concentration and exposure time. In vivo, the growth-inhibitory rate of crizotinib for H1993 xenografts was 72.3%. Positive expression rates of HGF and c-MET in H1993 xenografts were higher than those in A549 xenografts; the p-c-MET amount was the largest in H1993 xenograft control but the lowest in the H1993 xenograft with crizotinib treatment. The mRNA expression levels of c-MET and AKT in H1993 xenografts were higher than those of A549 xenografts. The protein levels of c-MET, AKT, and extracellular regulated protein kinases (ERK) in H1993 xenografts were higher than those in A549 xenografts; the p-AKT amount was higher in H1993 xenograft control than in A549 xenografts; the largest amount of p-c-MET was detected in H1993 xenograft control; the amount of p-ERK was the lowest in the H1993 xenograft with crizotinib treatment. Conclusion?The HGF/c-Met signaling pathway may mediate crizotinib-induced apoptosis and inhibition of proliferation of lung adenocarcinoma cells.
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