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Current Pharmaceutical Design

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ISSN (Print): 1381-6128
ISSN (Online): 1873-4286

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General Research Article

3-MA Enhanced Chemosensitivity in Cisplatin Resistant Hypopharyngeal Squamous Carcinoma Cells via Inhibiting Beclin -1 Mediated Autophagy

Author(s): Jia Zhang, Wei Mao, Yuying Liu, Jian Ding, Jie Wang, Ziwei Yu, Ruofei Huang, Sen Yang, Ying Sun and Pin Dong*

Volume 27, Issue 7, 2021

Published on: 21 December, 2020

Page: [996 - 1005] Pages: 10

DOI: 10.2174/1381612826666201221150431

Price: $65

Abstract

Background: Hypopharyngeal carcinoma is characterized by a high degree of malignancy. The most common pathological type is squamous cell carcinoma (HSCC). Cisplatin (cis-diamminedichloroplatinum, CDDP) is one of the most widely used chemotherapeutic drugs nowadays and cisplatin resistance is a major problem in current treatment strategies. Clinical researchers have reported that high autophagy levels often caused insensitivity to chemotherapy, a common phenomenon that greatly reduces the therapeutic effect in cisplatin- resistant tumor cell lines. 3-methyladenine (3-MA), an inhibitor of PI3K, plays a vital role in forming and developing autophagosomes. Therefore, we speculate that the use of 3-MA may reduce cisplatin resistance in hypopharyngeal squamous cell carcinoma (HSCC).

Methods: Part I: Cisplatin-resistant FaDu cell line (Human hypopharyngeal squamous cell carcinoma cells) was established and cultured. Cell counting kit-8 was used to detect drug resistance. An inverted microscope was used to observe the morphological changes at different concentrations, then the survival rate was calculated. After MDC staining, the autophagic vacuoles were observed by fluorescence microscopy. The expression of Beclin1 from each group was confirmed by RT-PCR and Western blot method. Part II: 3-MA was applied for cisplatin-resistant cells intervention, Beclin1 was knocked down by plasmid transfection. Cell cycle was detected using flow cytometry assay, apoptosis with necrosis was detected by staining with propidium iodide (PI). CCK-8 was used to observe the cell survival rate in each group. The expression of autophagy-related protein Beclin1, LC3I, LC3II, Atg-5 and P62 in each group was verified by Western blot analysis.

Results: Cisplatin-resistant FaDu cell line can be stably constructed by cisplatin intervention. Compared with normal group, autophagy and its related protein Beclin1 expression were enhanced in cisplatin resistant FaDu cells. Autophagy inhibition group showed significant cell cycle changes, mainly manifested by G1 arrest, increased apoptosis rate and significantly decreased survival rate at 24h level. The number of autophagy vacuoles were significantly reduced in the 3-MA group. Furthermore, Western blot showed that expression of Beclin1, lc3-I, lc3-II, atg-5 protein decreased significantly after 3-MA intervention, while the expression of p62 upregulated, which also confirmed autophagy flow was blocked.

Conclusion: Our work confirmed that enhanced autophagy is an important cause of cisplatin resistance in FaDu cells. The use of 3-MA can significantly reduce autophagy level and arresting its cell cycle, promote apoptosis and reverse the cisplatin resistance condition, this effect is partly mediated by inhibition of Beclin-1 expression. Our data provide a theoretical basis for the application of 3-MA in overcoming cisplatin resistance in hypopharyngeal cancer.

Keywords: 3-MA, HSCC, autophagy, cisplatin-resistance, Beclin 1, hypopharyngeal.

« Previous Next »
[1]
Bonner JA, Harari PM, Giralt J, et al. Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. N Engl J Med 2006; 354(6): 567-78.
[http://dx.doi.org/10.1056/NEJMoa053422] [PMID: 16467544]
[2]
Wang JH, Qi YF, Tang PZ, et al. Clinical analysis of posterior hypopharyngeal wall squamous cell carcinoma: a report of 36 cases. Chin J Cancer 2005; 24: 1106-10.
[3]
Ma J, Lu S, Yu L, et al. FaDu cell characteristics induced by multidrug resistance. Oncol Rep 2011; 26(5): 1189-95.
[PMID: 21833476]
[4]
Yang ZJ, Chee CE, Huang S, Sinicrope FA. The role of autophagy in cancer: therapeutic implications. Mol Cancer Ther 2011; 10(9): 1533-41.
[http://dx.doi.org/10.1158/1535-7163.MCT-11-0047] [PMID: 21878654]
[5]
Murtaza I, Saleem M, Adhami VM, Hafeez BB, Mukhtar H. Suppression of cFLIP by lupeol, a dietary triterpene, is sufficient to overcome resistance to TRAIL-mediated apoptosis in chemoresistant human pancreatic cancer cells. Cancer Res 2009; 69(3): 1156-65.
[http://dx.doi.org/10.1158/0008-5472.CAN-08-2917] [PMID: 19176377]
[6]
Keese M, Yagublu V, Schwenke K, Post S, Bastiaens P. Fluorescence lifetime imaging microscopy of chemotherapy-induced apoptosis resistance in a syngenic mouse tumor model. Int J Cancer 2010; 126(1): 104-13.
[http://dx.doi.org/10.1002/ijc.24730] [PMID: 19588498]
[7]
Mizushima N. Autophagy: process and function. Genes Dev 2007; 21(22): 2861-73.
[http://dx.doi.org/10.1101/gad.1599207] [PMID: 18006683]
[8]
Li J, Liu G, Li L, Yao Z, Huang J. Research progress on the effect of autophagy-lysosomal pathway on tumor drug resistance. Exp Cell Res 2020; 389(2)111925
[http://dx.doi.org/10.1016/j.yexcr.2020.111925]
[9]
Woo JS, Jung JS, Ha NC, et al. Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions. Cell Death Differ 2003; 10(12): 1310-9.
[http://dx.doi.org/10.1038/sj.cdd.4401303] [PMID: 12894216]
[10]
Akko Y, Berrak Z, Arsan ED, et al. Inhibition of PI3K signaling triggered apoptotic potential of curcumin which is hindered by Bcl-2 through activation of autophagy in MCF-7 cells. Biomedicine & Pharmacotherapy 71: 161-71.
[11]
Heathengel HM, Chang NC, Shore GC. The endoplasmic reticulum in apoptosis and autophagy: Role of the BCL-2 protein family 2008; 27: 6419-33.
[12]
Zhao J, Nie Y, Wang H, Lin Y. MiR-181a suppresses autophagy and sensitizes gastric cancer cells to cisplatin. Gene 2016; 576(2 Pt 2): 828-33.
[http://dx.doi.org/10.1016/j.gene.2015.11.013] [PMID: 26589846]
[13]
Tang ZH, Li T, Chang LL, et al. Glycyrrhetinic acid triggers a protective autophagy by activation of extracellular regulated protein kinases in hepatocellular carcinoma cells. J Agric Food Chem 2014; 62(49): 11910-6.
[http://dx.doi.org/10.1021/jf503968k] [PMID: 25403108]
[14]
Miller S, Oleksy A, Perisic O, Williams RL. Finding a fitting shoe for cinderella: searching for an autophagy inhibitor. Autophagy 2010; 6(6): 805-7.
[http://dx.doi.org/10.4161/auto.6.6.12577] [PMID: 20574157]
[15]
Zhang Y, Wang B, Chen X, Li W, Dong P. AGO2 involves the malignant phenotypes and FAK/PI3K/AKT signaling pathway in hypopharyngeal-derived FaDu cells. Oncotarget 2017; 8(33): 54735-46.
[http://dx.doi.org/10.18632/oncotarget.18047] [PMID: 28903378]
[16]
Le TD, Jin D, Rho SR, Kim MS, Yu R, Yoo H. Capsaicin-induced apoptosis of FaDu human pharyngeal squamous carcinoma cells. Yonsei Med J 2012; 53(4): 834-41.
[http://dx.doi.org/10.3349/ymj.2012.53.4.834] [PMID: 22665354]
[17]
Lang J, Gao L, Guo Y, et al. Comprehensive treatment of squamous cell cancer of head and neck: Chinese expert consensus 2013. Future Oncology 2013; 10: 1635-48.
[18]
Sewnaik A, Hoorweg JJ, Knegt PP, Wieringa MH, van der Beek JM, Kerrebijn JD. Treatment of hypopharyngeal carcinoma: analysis of nationwide study in the Netherlands over a 10-year period. Clin Otolaryngol 2005; 30(1): 52-7.
[http://dx.doi.org/10.1111/j.1365-2273.2004.00913.x] [PMID: 15748191]
[19]
Liu WS, Hsin CH, Chou YH, et al. Long-term results of intensity-modulated radiotherapy concomitant with chemotherapy for hypopharyngeal carcinoma aimed at laryngeal preservation. BMC Cancer 2010; 10: 102.
[http://dx.doi.org/10.1186/1471-2407-10-102] [PMID: 20298550]
[20]
Meinel FG, Mandl-Weber S, Baumann P, Leban J, Schmidmaier R. The novel, proteasome-independent NF-kappaB inhibitor V1810 induces apoptosis and cell cycle arrest in multiple myeloma and overcomes NF-kappaB-mediated drug resistance. Mol Cancer Ther 2010; 9(2): 300-10.
[http://dx.doi.org/10.1158/1535-7163.MCT-09-0645] [PMID: 20124446]
[21]
Janku F, McConkey DJ, Hong DS, Kurzrock R. Autophagy as a target for anticancer therapy. Nat Rev Clin Oncol 2011; 8(9): 528-39.
[http://dx.doi.org/10.1038/nrclinonc.2011.71] [PMID: 21587219]
[22]
Yang C, Gao R, Wang J, Yuan W, Wang C, Zhou X. High-mobility group nucleosome-binding domain 5 increases drug resistance in osteosarcoma through upregulating autophagy. Tumour Biol 2014; 35(7): 6357-63.
[http://dx.doi.org/10.1007/s13277-014-1833-0] [PMID: 24664583]
[23]
Mainz L, Rosenfeldt MT. Autophagy and cancer - insights from mouse models. FEBS J 2018; 285(5): 792-808.
[http://dx.doi.org/10.1111/febs.14274] [PMID: 28921866]
[24]
Bairey O, Vanichkin A, Shpilberg O. Arsenic-trioxide-induced apoptosis of chronic lymphocytic leukemia cells. Int J Lab Hematol 2010; 32(1 Pt 1): e77-85.
[http://dx.doi.org/10.1111/j.1751-553X.2008.01134.x] [PMID: 19208092]
[25]
Goussetis DJ, Platanias LC. Arsenic trioxide and the PI3K/AKT pathway in chronic lymphocytic leukemia. Clin Cancer Res 2010; 16: 4311.
[26]
Goussetis DJ, Altman JK, Glaser H, McNeer JL, Tallman MS, Platanias LC. Autophagy is a critical mechanism for the induction of the antileukemic effects of arsenic trioxide. J Biol Chem 2010; 285(39): 29989-97.
[http://dx.doi.org/10.1074/jbc.M109.090530] [PMID: 20656687]
[27]
Huang J, Ni J, Liu K, et al. HMGB1 Promotes drug resistance in osteosarcoma. Cancer Res 2012; 72(1): 230-8.
[28]
Sun X, Linyan MA, Zhang M, et al. Chloroquine induces apoptosis of human hepatocellular carcinoma cells in vitro by miR-26b-mediated regulation of Mcl-1 2018; 38: 409-13.
[29]
Song L, Liu H, Linyan MA, et al. Inhibition of autophagy by 3-MA enhances endoplasmic reticulum stress-induced apoptosis in human nasopharyngeal carcinoma cells. Oncot Lett 2013; 6: 1031-8.
[30]
Guo S, Yang B, Liu H, et al. Experimental study of autophagy inhibitor 3-MA combined with PI3K/mTOR dual inhibitor NVP-BEZ235 promote apoptosis of cervical cancer cells. J Youjiang Medical University Nationalities 2019.
[31]
Denecker G, Vercammen D, Declercq W, et al. Apoptotic and necrotic cell death induced by death domain receptors. Cell Mol Life Sci 58: 356-70.

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