Identification of a novel autophagic inhibitor cepharanthine to enhance the anti-cancer property of dacomitinib in non-small cell lung cancer

Zheng-hai TANG,Wen-xiang CAO,Xia GUO,Xiao-yang DAI,Jia-hong LU,Xiu-ping CHEN,Hong ZHU,Jin-jian LU

(1.State Key Laboratory of Quality Research in Chinese Medicine,Institute of Chinese Medical Sciences,University of Macau,Macao,China;2.Zhejiang Province Key Laboratory of Anti-cancer Drug Research,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China）

T2-18

Identification of a novel autophagic inhibitor cepharanthine to enhance the anti-cancer property of dacomitinib in non-small cell lung cancer

Zheng-hai TANG1,Wen-xiang CAO1,Xia GUO1,Xiao-yang DAI2,Jia-hong LU1,Xiu-ping CHEN1,Hong ZHU2,Jin-jian LU1

(1.State Key Laboratory of Quality Research in Chinese Medicine,Institute of Chinese Medical Sciences,University of Macau,Macao,China;2.Zhejiang Province Key Laboratory of Anti-cancer Drug Research,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China）

OBJECTIVEIdentification of novel autophagy inhibitors for the combinational treatment of non-small cell lung cancer(NSCLC).METHODSMTT assay and annexin V/PI staining assay were used to evaluate the cell proliferation and apoptosis,respectively.Immunofluorescence staining and cathepsin activity assay were used to detect autophagy.Small interfering RNA was performed to silence the genes and Western blot assay was used to evaluate the protein express levels.Xenograft experiments were applied for in vivo evaluation.RESULTSCepharanthine,a natural compound,increased LC3-II expression and GFP-LC3 puncta formation in NSCLC NCI-H1975 cells.Numerous yellow puncta were observed in cepharanthine-treated cells with mRFP-EGFP-LC3 transfection.Co-staining of GFP-LC3 with LysoTracker red or LAMP1 antibody suggested that cepharanthine inhibits autophagosomes-lysosomes fusion.Moreover,cepharanthine attenuated the lysosomal cathepsins maturation.Wealsoconfirmedthatdacomitinibinducedcytoprotectiveautophagy.Combined treatment with cepharanthine increased the anti-cancer effects of dacomitinib in vitro and in vivo.Besides,cepharanthine could not enhance the anti-cancer effect of dacomitinib in autophagy deficient cells.CONCLUSIONCepharanthine might be further developed as a promising autophagic inhibitor,and combined treatment cepharanthine with dacomitinib could pose as an effective strategy for NSCLC treatment.

cepharanthine;non-small cell lung cancer;autophagy;lysosome;dacomitinib

The project supported by Science and Technology Development Fund,Macao S.A.R(FDCT)(024/2016/A1),and Research Fund of University of Macau(MYRG2015-00091-ICMS-QRCM and MYRG2015-00101-ICMS-QRCM)

Jin-jian LU,E-mail:jinjianlu@umac.mo;jinjian.lu@163.com