THADA inhibits autophagy and increases 5-FU sensitivity in gastric cancer cells via the PI3K/AKT/mTOR signaling pathway

Document Type : Original Article

Authors

1 Department of Gastrointestinal Surgery, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

2 Department of Hematology, The First Affiliated Hospital, Zhejiang University College of Medicine, Hangzhou, China

3 Zhejiang University College of Medicine, Hangzhou, China

Abstract

Objective(s): 5-Fluorouracil (5-FU) is currently the main drug used in chemotherapy for gastric cancer (GC). The main clinical problems of 5-FU therapy are insensitivity and acquired resistance to 5-FU. The mechanism of GC cell resistance to 5-FU is currently unknown.
Materials and Methods: This study employed next-generation sequencing (NGS) to analyze the differentially expressed genes (DEGs) in chemotherapy-sensitive and non-sensitive GC tissues. In addition, a bioinformatics analysis was conducted using the GC dataset of GEO, and further validated and explored through in vitro experiments.
Results: Thyroid adenoma-associated gene (THADA) was highly expressed in GC tissues from chemotherapy-sensitive patients and was an independent prognostic factor in GC patients receiving postoperative 5-FU adjuvant chemotherapy. Notably, heightened THADA expression in GC cells was associated with the down-regulation of autophagy-related proteins (LC-3, ATG13, ULK1, and TFEB). Furthermore, the PI3K/AKT/mTOR signaling pathway and mTORC1 signaling pathway were remarkably increased in patients with elevated THADA expression. THADA expression was associated with mTOR, the core protein of the mTOR signaling pathway, and related proteins involved in regulating the mTORC1 signaling pathway (mLST8, RHEB, and TSC2). THADA exhibited inhibitory effects on autophagy and augmented the sensitivity of GC cells to 5-FU through the PI3K/AKT/mTOR signaling pathway.
Conclusion: The findings suggest that THADA may be involved in the regulatory mechanism of GC cell sensitivity to 5-FU. Consequently, the detection of THADA in tumor tissues may bring clinical benefits, specifically for 5-FU-related chemotherapy administered to GC patients with elevated THADA expression.

Keywords

Main Subjects

References

1. Sung H, Ferlay J, Siegel R L, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021;71:209-249.
2. Ajani J A, D’Amico T A, Bentrem D J, Chao J, Cooke D, Corvera C, et al. Gastric Cancer, Version 2.2022, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw 2022;20:167-192.
3. Ma J, Shen H, Kapesa L, Zeng S. Lauren classification and individualized chemotherapy in gastric cancer. Oncol Lett 2016; 11:2959-2964.
4. Vodenkova S, Buchler T, Cervena K, Veskrnova V, Vodicka P, Vymetalkova V. 5-fluorouracil and other fluoropyrimidines in colorectal cancer: Past, present and future. Pharmacol Ther 2020; 206:107447.
5. Machover D, Goldschmidt E, Chollet P, Metzger G, Zittoun J, Benavides M, et al. Treatment of advanced colorectal and gastric adenocarcinomas with 5-fluorouracil and high-dose folinic acid. NCI Monogr 1987;5:193-198.
6. Sethy C, Kundu C N. 5-Fluorouracil (5-FU) resistance and the new strategy to enhance the sensitivity against cancer: Implication of DNA repair inhibition. Biomed Pharmacother 2021;137:111285.
7. Mizushima N. Autophagy: process and function. Genes Dev 2007; 21:2861-2873.
8. Jiang G M, Tan Y, Wang H, Peng L, Chen H T, Meng X J, et al. The relationship between autophagy and the immune system and its applications for tumor immunotherapy. Mol Cancer, 2019;18:17-38.
9. Collado M, Blasco M A, Serrano M. Cellular senescence in cancer and aging. Cell 2007;130:223-233.
10. Sun Y, Coppé J P, Lam E W. Cellular senescence: The sought or the unwanted? Trends Mol Med 2018;24:871-885.
11. Xing M. Genetic alterations in the phosphatidylinositol-3 kinase/Akt pathway in thyroid cancer. Thyroid 2010; 20: 697-706.
12. Spirina LV, Usynin YA, Yurmazov ZA, Slonimskaya EM, Kolegova ES, Kondakova IV. [Transcription factors NF-kB, HIF-1, HIF-2, growth factor VEGF, VEGFR2 and carboanhydrase IX mRNA and protein level in the development of kidney cancer metastasis]. Mol Biol (Mosk) 2017;51:372-377.
13. Ying LS, Yu JL, Lu XX, Ling ZQ. Enhanced RegIV expression predicts the intrinsic 5-fluorouracil (5-FU) resistance in advanced gastric cancer. Dig Dis Sci 2013;58: 414-422.
14. Cardona A, Pagani L, Antao T, Lawson D J, Eichstaedt C A, Yngvadottir B, et al. Genome-wide analysis of cold adaptation in indigenous Siberian populations. PLoS One 2014;9:e98076-e98086.
15. Moraru A, Cakan-Akdogan G, Strassburger K, Males M, Mueller S, Jabs M, et al. THADA Regulates the Organismal Balance between Energy Storage and Heat Production. Dev Cell 2017;41:72-8.
16. Prasad RB, Lessmark A, Almgren P, Kovacs G, Hansson O, Oskolkov N, et al. Excess maternal transmission of variants in the THADA gene to offspring with type 2 diabetes. Diabetologia 2016;59:1702-1713.
17. Goodarzi MO, Jones MR, Li X, Chua AK, Garcia OA, Chen YD, et al. Replication of association of DENND1A and THADA variants with polycystic ovary syndrome in European cohorts. J Med Genet 2012;49: 90-95.
18. Rippe V, Drieschner N, Meiboom M, Murua Escobar H, Bonk U, Belge G, et al. Identification of a gene rearranged by 2p21 aberrations in thyroid adenomas. Oncogene 2003;22: 6111-6114.
19. Cheng I, Caberto CP, Lum-Jones A, Seifried A, Wilkens LR, Schumacher FR, et al. Type 2 diabetes risk variants and colorectal cancer risk: The multiethnic cohort and PAGE studies. Gut 2011;60:1703-1711.
20. Panebianco F, Kelly LM, Liu P, Zhong S, Dacic S, Wang X, et al. THADA fusion is a mechanism of IGF2BP3 activation and IGF1R signaling in thyroid cancer. Proc Natl Acad Sci U S A 2017;114:2307-2312.
21. Nikiforov Y E, Seethala R R, Tallini G, Baloch Z W, Basolo F, Thompson L D, et al. Nomenclature revision for encapsulated follicular variant of papillary thyroid carcinoma: A paradigm shift to reduce overtreatment of indolent tumors. JAMA Oncol 2016;2:1023-1029.
22. Li C, Chi H, Deng S, Wang H, Yao H, Wang Y, et al. THADA drives Golgi residency and upregulation of PD-L1 in cancer cells and provides promising target for immunotherapy. J Immunother Cancer 2021;9:e002443-2459. 
23. Nannini M, Pantaleo M A, Maleddu A, Astolfi A, Formica S, Biasco G. Gene expression profiling in colorectal cancer using microarray technologies: Results and perspectives. Cancer Treat Rev 2009;35:201-209.
24. Kulasingam V, Diamandis EP. Strategies for discovering novel cancer biomarkers through utilization of emerging technologies. Nat Clin Pract Oncol 2008;5:588-599.
25. Barnard RA, Regan DP, Hansen RJ, Maycotte P, Thorburn A, Gustafson DL. Autophagy Inhibition delays early but not late-stage metastatic disease. J Pharmacol Exp Ther 2016;358:282-293.
26. Guo J Y, Xia B, White E. Autophagy-mediated tumor promotion. Cell 2013;155:1216-1219.
27. Liu F, Ai FY, Zhang DC, Tian L, Yang ZY, Liu SJ. LncRNA NEAT1 knockdown attenuates autophagy to elevate 5-FU sensitivity in colorectal cancer via targeting miR-34a. Cancer Med 2020;9:1079-1091.
28. Feng Y, Gao Y, Wang D, Xu Z, Sun W, Ren P. Autophagy inhibitor (LY294002) and 5-fluorouracil (5-FU) combination-based nanoliposome for enhanced efficacy against esophageal squamous cell carcinoma. Nanoscale Res Lett 2018;13:325-333.
29. Yadunandam AK, Yoon JS, Seong YA, Oh CW, Kim GD. Prospective impact of 5-FU in the induction of endoplasmic reticulum stress, modulation of GRP78 expression and autophagy in Sk-Hep1 cells. Int J Oncol 2012;41:1036-1042.
30. Li L Q, Xie W J, Pan D, Chen H, Zhang L. Inhibition of autophagy by bafilomycin A1 promotes chemosensitivity of gastric cancer cells. Tumour Biol 2016;37:653-659.
31. Bhattacharya B, Low SH, Soh C, Kamal Mustapa N, Beloueche-Babari M, Koh KX, et al. Increased drug resistance is associated with reduced glucose levels and an enhanced glycolysis phenotype. Br J Pharmacol 2014;171:3255-3267.
32. Yuan L, Xu ZY, Ruan SM, Mo S, Qin JJ, Cheng XD. Long non-coding RNAs towards precision medicine in gastric cancer: Early diagnosis, treatment, and drug resistance. Mol Cancer 2020;19: 96-117.
33. Fang L, Lv J, Xuan Z, Li B, Li Z, He Z, et al. Circular CPM promotes chemoresistance of gastric cancer via activating PRKAA2-mediated autophagy. Clin Transl Med 2022;12:e708-725.
34. Zhu BS, Sun JL, Gong W, Zhang XD, Wu YY, and Xing CG. Effects of 5‑fluorouracil and class III phosphoinositide 3‑kinase small interfering RNA combination therapy on SGC7901 human gastric cancer cells. Mol Med Rep 2015;11:1891-1898.
35. Dikic I, Elazar Z. Mechanism and medical implications of mammalian autophagy. Nat Rev Mol Cell Biol 2018;19: 349-364.
36. Kim J, Kundu M, Viollet B, Guan KL. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 2011;13:132-141.
37. Settembre C, Di Malta C, Polito VA, Garcia Arencibia M, Vetrini F, Erdin S, et al. TFEB links autophagy to lysosomal biogenesis. Science 2011;332: 1429-1433.
38. Hanahan D, Weinberg RA. Hallmarks of cancer: The next generation. Cell 2011;144: 646-674.
39. Krencz I, Sebestyen A, Papay J, Jeney A, Hujber Z, Burger CD, et al. In situ analysis of mTORC1/2 and cellular metabolism-related proteins in human Lymphangioleiomyomatosis. Hum Pathol 2018; 79:199-207.
40. Cotler SJ, Hay N, Xie H, Chen ML, Xu PZ, Layden TJ, et al. Immunohistochemical expression of components of the Akt-mTORC1 pathway is associated with hepatocellular carcinoma in patients with chronic liver disease. Dig Dis Sci 2008;53: 844-849.
41. Faried LS, Faried A, Kanuma T, Aoki H, Sano T, Nakazato T, et al. Expression of an activated mammalian target of rapamycin in adenocarcinoma of the cervix: A potential biomarker and molecular target therapy. Mol Carcinog 2008;47: 446-457.
42. Lee H. Phosphorylated mTOR Expression Profiles in Human Normal and Carcinoma Tissues. Dis Markers 2017;2017:1397063-1397070.
43. Jiang WJ, Feng RX, Liu JT, Fan LL, Wang H, Sun GP. RICTOR expression in esophageal squamous cell carcinoma and its clinical significance. Med Oncol 2017;34: 32.
44. Pópulo H, Lopes JM, Soares P. The mTOR signalling pathway in human cancer. Int J Mol Sci 2012. 13: 1886-1918.
45. Mossmann D, Park S, Hall MN. mTOR signalling and cellular metabolism are mutual determinants in cancer. Nat Rev Cancer 2018;18: 744-757.
46. Yang H, Rudge DG, Koos JD, Vaidialingam B, Yang HJ, Pavletich NP. mTOR kinase structure, mechanism and regulation. Nature 2013;497:217-223.
47. Rogala KB, Gu X, Kedir JF, Abu-Remaileh M, Bianchi LF, Bottino AMS, et al. Structural basis for the docking of mTORC1 on the lysosomal surface. Science 2019;366:468-475.
48. Jewell JL, Russell RC, Guan KL. Amino acid signalling upstream of mTOR. Nat Rev Mol Cell Biol 2013;14:133-139.
Iranian Journal of Basic Medical Sciences
Volume 27, Issue 2
February 2024
Pages 195-202

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