Anlotinib combined with chloroquine induces apoptosis of human non-small cell lung cancer cell line H1299

Abstract

Abstract: Objective To investigate the effects of anlotinib and chloroquine (CQ) on viability, invasion and apoptosis of H1299 cells. Methods After treatment of H1299 cells with anlotinib or/and CQ, the cell viability was measured by MTT assay; Wound-healing and Transwell chamber assay were used to evaluate cell migration and invasion abilities; Hoechst 33342 staining was adopted to test the changes of apoptosis; The protein levels of autophagy marker LC3-Ⅱ/LC3-Ⅰ and autophagic substrate p62 were detected by Western blot. Results Anlotinib combined with CQ showed more obviously synergistic inhibitory effect on the viability, invasion and migration in H1299 cells than anlotinib alone (P＜0.05). As compared to the untreated control cells, anlotinib alone promoted the cell apoptosis. Their combination induced an even higher apoptosis in H1299 cells. With the increase of concentration, anlotinib significantly up-regulated autophagy marker protein LC3-Ⅱ/LC3-Ⅰ and down-regulated autophagic substrate protein p62.The expression of LC3-Ⅱ/LC3-Ⅰ and p62 was significantly up-regulated by CQ. Compared with the CQ group alone, LC3-Ⅱ/LC3-Ⅰ and p62 were significantly up-regulated and down-regulated in the 20 μmol/L anlotinib combined with CQ group. Conclusions Combination of anlotinib and CQ remarkably inhibits the viability, migration and invasion and induces the apoptosis of H1299 cells. The potential mechanism for antitumor might be due to autophagy's action.

Key words: non-small cell lung cancer(NSCLC), anlotinib, chloroquine, autophagy, apoptosis

CLC Number:

R734.2

LIU Xi-bang, ZHU Lin-zhi, JIAO De-min, TANG Xia-li, CHEN Jun, HU Xu-gang, CHEN Qing-yong. Anlotinib combined with chloroquine induces apoptosis of human non-small cell lung cancer cell line H1299[J]. Basic & Clinical Medicine, 2021, 41(7): 1018-1023.

References

[1]Reck M, Rabe KF. Precision diagnosis and treatment for advanced non-small-cell lung cancer[J]. N Engl J Med, 2017, 377: 849-861.
[2]Lo Sardo F, Strano S, Blandino G. YAP and TAZ in lung cancer: oncogenic role and clinical targeting[J]. Cancers (Basel), 2018, 10: 137-166.
[3]Lin B, Song X, Yang D, et al. Anlotinib inhibits angiogenesis via suppressing the activation of VEGFR2, PDGFR beta and FGFR1[J]. Gene, 2018, 654: 77-86.
[4]Han B, Li K, Wang Q, et al. Effect of anlotinib as a third-line or further treatment on overall survival of patients with advanced non-small cell lung cancer: the ALTER 0303 phase 3 randomized clinical trial[J]. JAMA Oncol, 2018, 4: 1569-1575.
[5]Lu J, Zhong H, Chu T, et al. Role of anlotinib-induced CCL2 decrease in anti-angiogenesis and response prediction for nonsmall cell lung cancer therapy[J]. Eur Respir J, 2019, 53: 1801562. doi: 10.1183/13993003.01562-2018.
[6]Santana-Codina N, Mancias JD, Kimmelman AC. The role of autophagy in cancer[J]. Annu Rev Cancer Biol, 2017, 1: 19-39.
[7]Levy JMM, Towers CG, Thorburn A. Targeting autophagy in cancer[J]. Nat Rev Cancer, 2017, 17: 528-542.
[8]Kinsey CG, Camolotto SA, Boespflug AM, et al. Protec-tive autophagy elicited by RAF->MEK->ERK inhibition suggests a treatment strategy for RAS-driven cancers[J]. Nat Med, 2019, 25: 620-627.
[9]Chi Y, Yao Y, Wang SS, et al. Anlotinib for metastasis soft tissue sarcoma: a randomized, double-blind, placebo-controlled and multi-centered clinical trial[J]. J Clin Oncol, 2018, 36: 11503.
[10]Li DP, Tang PZ, Chen XH, et al. Anlotinib treatment in locally advanced or metastatic medullary thyroid carcinoma: a multicenter, randomized, double-blind, placebo-controlled phase IIB trial[J]. J Clin Oncol, 2019, 37: 6019.
[11]Si X, Zhang L, Wang H, et al. Management of anlotinib-related adverse events in patients with advanced non-small cell lung cancer: experiences in ALTER-0303[J]. Thorac Cancer, 2019, 10: 551-556.
[12]Chou HL, Lin YH, Liu W, et al. Combination therapy of chloroquine and C(2)-ceramide enhances cytotoxicity in lung cancer H460 and H1299 cells[J]. Cancers (Basel), 2019, 11: 370-386.
[13]Fujii S, Hara H, Araya J, et al. Insufficient autophagy promotes bronchial epithelial cell senescence in chronic obstructive pulmonary disease[J]. Oncoimmunology, 2012, 1: 630-641.
[14]Redmann M, Benavides GA, Berryhill TF, et al. Inhibi-tion of autophagy with bafilomycin and chloroquine decreases mitochondrial quality and bioenergetic function in primary neurons[J]. Redox Biol, 2017, 11: 73-81.

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