miR‑4486 reverses cisplatin‑resistance of colon cancer cells via targeting ATG7 to inhibiting autophagy

Wang,Weiwei; Chen,Linxia; Zhu,Wenjin; Huang,Xianjin; Lin,Lin; Quan,Zhenhao; Sun,Kaiyu; Xu,Qingwen

doi:10.3892/etm.2021.10900

miR‑4486 reverses cisplatin‑resistance of colon cancer cells via targeting ATG7 to inhibiting autophagy

Authors:
- Weiwei Wang
- Linxia Chen
- Wenjin Zhu
- Xianjin Huang
- Lin Lin
- Zhenhao Quan
- Kaiyu Sun
- Qingwen Xu
View Affiliations

Affiliations: Department of Gastrointestinal Surgery, The Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524000, P.R. China, Department of Operating Room, The Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524000, P.R. China
Published online on: October 20, 2021 https://doi.org/10.3892/etm.2021.10900
Article Number: 1465
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cisplatin (DDP) resistance is one of the main causes of treatment failure in patients with colon cancer (CC). Autophagy is a key mechanism of resistance to chemotherapy. Since autophagy‑related 7 (ATG7) has been reported to be involved in the regulation of autophagy and DDP resistance for lung and esophageal cancer, the present study aimed to explore the functions of microRNA (miR)‑4486 in the autophagy‑mediated DDP resistance of CC. The expression level of miR‑4486 in HCT116, DDP‑resistant HCT116 cells (HCT116/DDP), SW480 and DDP‑resistant SW480 cells (SW480/DDP) was quantified by reverse transcription‑quantitative PCR. Western blotting was utilized to analyze the expression of ATG7, autophagy‑related proteins Beclin 1 and LC3‑I/II, as well as apoptosis‑related proteins Bcl‑2, Bax and cleaved‑caspase 3 in HCT116/DDP and SW480/DDP cells. The half maximal inhibitory concentration of DDP on all cell lines and the cell viability of HCT116/DDP and SW480/DDP cells were measured using Cell Counting Kit 8 assay. Luciferase assay was used to examine the potential targets of miR‑4486 and ATG7. The effects of upregulating mimic miR‑4486 expression on the apoptosis and autophagy of HCT116/DDP and SW480/DDP cells were determined by flow cytometry and electron microscopy, respectively. It was found that miR‑4486 expression was significantly decreased in HCT116/DDP and SW480/DDP cells compared with that in HCT116 and SW480 cells. Overexpression of miR‑4486 could increase the sensitivity of HCT116/DDP and SW480/DDP cells to DDP by reducing cell viability, promoting apoptosis and inhibiting autophagy through downregulating Beclin 1 expression and the LC3‑II/LC3‑I ratio. Additionally, ATG7 was identified to be a target gene of miR‑4486, where ATG7 overexpression could partially reverse the effects of miR‑4486 on cell viability and apoptosis by promoting the formation of autophagosomes. In conclusion, the present results demonstrated that miR‑4486 could reverse DDP resistance in HCT116/DDP and SW480/DDP cells by targeting ATG7 to inhibit autophagy.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018.PubMed/NCBI View Article : Google Scholar
2	Liu N, Wu C, Jia R, Cai G, Wang Y, Zhou L, Ji Q, Sui H, Zeng P, Xiao H, et al: Traditional Chinese medicine combined with chemotherapy and cetuximab or bevacizumab for metastatic colorectal cancer: A randomized, double-blind, placebo-controlled clinical trial. Front Pharmacol. 11(478)2020.PubMed/NCBI View Article : Google Scholar
3	Paldino E, Tesori V, Casalbore P, Gasbarrini A and Puglisi MA: Tumor initiating cells and chemoresistance: Which is the best strategy to target colon cancer stem cells? Biomed Res Int. 2014(859871)2014.PubMed/NCBI View Article : Google Scholar
4	Duffy A, Le J, Sausville E and Emadi A: Autophagy modulation: A target for cancer treatment development. Cancer Chemother Pharmacol. 75:439–447. 2015.PubMed/NCBI View Article : Google Scholar
5	Sui X, Chen R, Wang Z, Huang Z, Kong N, Zhang M, Han W, Lou F, Yang J, Zhang Q, et al: Autophagy and chemotherapy resistance: A promising therapeutic target for cancer treatment. Cell Death Dis. 4(e838)2013.PubMed/NCBI View Article : Google Scholar
6	Devenport SN and Shah YM: Functions and implications of autophagy in colon cancer. Cells. 8(1349)2019.PubMed/NCBI View Article : Google Scholar
7	Di Leva G and Croce CM: miRNA profiling of cancer. Curr Opin Genet Dev. 23:3–11. 2013.PubMed/NCBI View Article : Google Scholar
8	Aguda BD: Modeling microRNA-transcription factor networks in cancer. Adv Exp Med Biol. 774:149–167. 2013.PubMed/NCBI View Article : Google Scholar
9	Baer C, Claus R and Plass C: Genome-wide epigenetic regulation of miRNAs in cancer. Cancer Res. 73:473–477. 2013.PubMed/NCBI View Article : Google Scholar
10	Song B, Wang Y, Xi Y, Kudo K, Bruheim S, Botchkina GI, Gavin E, Wan Y, Formentini A, Kornmann M, et al: Mechanism of chemoresistance mediated by miR-140 in human osteosarcoma and colon cancer cells. Oncogene. 28:4065–4074. 2009.PubMed/NCBI View Article : Google Scholar
11	Chai H, Liu M, Tian R, Li X and Tang H: miR-20a targets BNIP2 and contributes chemotherapeutic resistance in colorectal adenocarcinoma SW480 and SW620 cell lines. Acta Biochim Biophys Sin (Shanghai). 43:217–225. 2011.PubMed/NCBI View Article : Google Scholar
12	Guo Y, Pang Y, Gao X, Zhao M, Zhang X, Zhang H, Xuan B and Wang Y: MicroRNA-137 chemosensitizes colon cancer cells to the chemotherapeutic drug oxaliplatin (OXA) by targeting YBX1. Cancer Biomarkers. 18:1–9. 2017.PubMed/NCBI View Article : Google Scholar
13	Qu J, Zhao L, Zhang P, Wang J, Xu N, Mi W, Jiang X, Zhang C and Qu J: MicroRNA-195 chemosensitizes colon cancer cells to the chemotherapeutic drug doxorubicin by targeting the first binding site of BCL2L2 mRNA. J Cell Physiol. 230:535–545. 2015.PubMed/NCBI View Article : Google Scholar
14	Liu J, Cheng LG and Li HG: LncRNA SNHG20 promoted the proliferation of glioma cells via sponging miR-4486 to regulate the MDM2-p53 pathway. Eur Rev Med Pharmacol Sci. 23:5323–5331. 2019.PubMed/NCBI View Article : Google Scholar
15	Asensio VJ, Tomás A, Iglesias A, de Llano LP, Del Pozo V and Cosío BG: CHACOS study group. Eosinophilic COPD patients display a distinctive serum mirna profile from asthma and non-eosinophilic COPD. Arch Bronconeumol (Engl ED). 56:234–241. 2020.PubMed/NCBI View Article : Google Scholar : (In English, Spanish).
16	Kim SH, Yun SW, Kim HR and Chae SA: Exosomal microRNA expression profiles of cerebrospinal fluid in febrile seizure patients. Seizure. 81:47–52. 2020.PubMed/NCBI View Article : Google Scholar
17	Hao H, Xia G, Wang C, Zhong F, Liu L and Zhang D: miR-106a suppresses tumor cells death in colorectal cancer through targeting ATG7. Med Mol Morphol. 50:76–85. 2017.PubMed/NCBI View Article : Google Scholar
18	Xie Q, Liu Y and Li X: The interaction mechanism between autophagy and apoptosis in colon cancer. Transl Oncol. 13(100871)2020.PubMed/NCBI View Article : Google Scholar
19	Benson AB III, Venook AP, Cederquist L, Chan E, Chen YJ, Cooper HS, Deming D, Engstrom PF, Enzinger PC, Fichera A, et al: Colon cancer, version 1.2017, NCCN clinical practice guidelines in oncology. J Natl Compr Canc Netw. 15:370–398. 2017.PubMed/NCBI View Article : Google Scholar
20	Bosman FT, Carneiro F, Hruban RH and Theise ND (eds.): WHO Classification of Tumors of the Digestive System. Vol 3. 4th edition. IARC Press, Lyon, 2010.
21	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
22	Song L, Zhou Z, Gan Y, Li P, Xu Y, Zhang Z, Luo F, Xu J, Zhou Q and Dai F: Long noncoding RNA OIP5-AS1 causes cisplatin resistance in osteosarcoma through inducing the LPAATbeta/PI3K/AKT/mTOR signaling pathway by sponging the miR-340-5p. J Cell Biochem. 120:9656–9666. 2019.PubMed/NCBI View Article : Google Scholar
23	Pan B, Chen Y, Song H, Xu Y, Wang R and Chen L: Mir-24-3p downregulation contributes to VP16-DDP resistance in small-cell lung cancer by targeting ATG4A. Oncotarget. 6:317–331. 2015.PubMed/NCBI View Article : Google Scholar
24	Lin C, Xie L, Lu Y, Hu Z and Chang J: miR-133b reverses cisplatin resistance by targeting GSTP1 in cisplatin-resistant lung cancer cells. Int J Mol Med. 41:2050–2058. 2018.PubMed/NCBI View Article : Google Scholar
25	Qin X, Guo H, Wang X, Zhu X, Yan M, Wang X, Xu Q, Shi J, Lu E, Chen W and Zhang J: Exosomal miR-196a derived from cancer-associated fibroblasts confers cisplatin resistance in head and neck cancer through targeting CDKN1B and ING5. Genome Biol. 20(12)2019.PubMed/NCBI View Article : Google Scholar
26	Chan H, Pearson CS, Green CM, Li Z, Zhang J, Belfort G, Shekhtman A, Li H and Belfort M: Exploring intein inhibition by platinum compounds as an antimicrobial strategy. J Biol Chem. 291:22661–22670. 2016.PubMed/NCBI View Article : Google Scholar
27	Dasari S and Tchounwou PB: Cisplatin in cancer therapy: Molecular mechanisms of action. Eur J Pharmacol. 740:364–378. 2014.PubMed/NCBI View Article : Google Scholar
28	Sun CY, Zhang QY, Zheng GJ and Feng B: Phytochemicals: Current strategy to sensitize cancer cells to cisplatin. Biomed Pharmacother. 110:518–527. 2019.PubMed/NCBI View Article : Google Scholar
29	Siddik ZH: Cisplatin: Mode of cytotoxic action and molecular basis of resistance. Oncogene. 22:7265–7279. 2003.PubMed/NCBI View Article : Google Scholar
30	Oun R, Moussa YE and Wheate NJ: Correction: The side effects of platinum-based chemotherapy drugs: A review for chemists. Dalton Trans. 47(7848)2018.PubMed/NCBI View Article : Google Scholar
31	Köberle B, Tomicic MT, Usanova S and Kaina B: Cisplatin resistance: Preclinical findings and clinical implications. Biochim Biophys Acta. 1806:172–182. 2010.PubMed/NCBI View Article : Google Scholar
32	Wu WK, Coffelt SB, Cho CH, Wang XJ, Lee CW, Chan FK, Yu J and Sung JJ: The autophagic paradox in cancer therapy. Oncogene. 31:939–953. 2012.PubMed/NCBI View Article : Google Scholar
33	Wang K: Autophagy and apoptosis in liver injury. Cell Cycle. 14:1631–1642. 2015.PubMed/NCBI View Article : Google Scholar
34	Bursch W: The autophagosomal-lysosomal compartment in programmed cell death. Cell Death Differ. 8:569–581. 2001.PubMed/NCBI View Article : Google Scholar
35	Shimizu S, Kanaseki T, Mizushima N, Mizuta T, Arakawa-Kobayashi S, Thompson CB and Tsujimoto Y: Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes. Nat Cell Biol. 6:1221–1228. 2004.PubMed/NCBI View Article : Google Scholar
36	He C and Levine B: The beclin 1 interactome. Curr Opin Cell Biol. 22:140–149. 2010.PubMed/NCBI View Article : Google Scholar
37	Itakura E, Kishi C, Inoue K and Mizushima N: Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG. Mol Biol Cell. 19:5360–5372. 2008.PubMed/NCBI View Article : Google Scholar
38	Deretic V: Autophagosome and phagosome. Methods Mol Biol. 445:1–10. 2008.PubMed/NCBI View Article : Google Scholar
39	Fu Ll, Cheng Y and Liu B: Beclin-1: Autophagic regulator and therapeutic target in cancer. Int J Biochem Cell Biol. 45:921–924. 2013.PubMed/NCBI View Article : Google Scholar
40	Noguchi M, Hirata N, Tanaka T, Suizu F, Nakajima H and Chiorini JA: Autophagy as a modulator of cell death machinery. Cell Death Dis. 11(517)2020.PubMed/NCBI View Article : Google Scholar
41	Kang R, Zeh HJ, Lotze MT and Tang D: The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ. 18:571–580. 2011.PubMed/NCBI View Article : Google Scholar
42	Guo GF, Wang YX, Zhang YJ, Chen XX, Lu JB, Wang HH, Jiang C, Qiu HQ and Xia LP: Predictive and prognostic implications of 4E-BP1, Beclin-1, and LC3 for cetuximab treatment combined with chemotherapy in advanced colorectal cancer with wild-type KRAS: Analysis from real-world data. World J Gastroenterol. 25:1840–1853. 2019.PubMed/NCBI View Article : Google Scholar
43	Guo JY, Xia B and White E: Autophagy-mediated tumor promotion. Cell. 155:1216–1219. 2013.PubMed/NCBI View Article : Google Scholar
44	Bellodi C, Lidonnici MR, Hamilton A, Helgason GV, Soliera AR, Ronchetti M, Galavotti S, Young KW, Selmi T, Yacobi R, et al: Targeting autophagy potentiates tyrosine kinase inhibitor-induced cell death in Philadelphia chromosome-positive cells, including primary CML stem cells. J Clin Invest. 119:1109–1123. 2009.PubMed/NCBI View Article : Google Scholar
45	Li H, Jin X, Zhang Z, Xing Y and Kong X: Inhibition of autophagy enhances apoptosis induced by the PI3K/AKT/mTor inhibitor NVP-BEZ235 in renal cell carcinoma cells. Cell Biochem Funct. 31:427–433. 2013.PubMed/NCBI View Article : Google Scholar
46	Lin CJ, Lee CC, Shih YL, Lin TY, Wang SH, Lin YF and Shih CM: Resveratrol enhances the therapeutic effect of temozolomide against malignant glioma in vitro and in vivo by inhibiting autophagy. Free Radic Biol Med. 52:377–391. 2012.PubMed/NCBI View Article : Google Scholar
47	Chen X, Xu H, Yu X, Wang X, Zhu X and Xu X: Apigenin inhibits in vitro and in vivo tumorigenesis in cisplatin-resistant colon cancer cells by inducing autophagy, programmed cell death and targeting m-TOR/PI3K/Akt signalling pathway. J BUON. 24:488–493. 2019.PubMed/NCBI
48	Tan S, Shi H, Ba M, Lin S, Tang H, Zeng X and Zhang X: miR-409-3p sensitizes colon cancer cells to oxaliplatin by inhibiting Beclin-1-mediated autophagy. Int J Mol Med. 37:1030–1038. 2016.PubMed/NCBI View Article : Google Scholar
49	He Y, Zhang L, Tan F, Wang LF, Liu DH, Wang RJ and Yin XZ: MiR-153-5p promotes sensibility of colorectal cancer cells to oxaliplatin via targeting Bcl-2-mediated autophagy pathway. Biosci Biotechnol Biochem. 84:1645–1651. 2020.PubMed/NCBI View Article : Google Scholar
50	Cernigliaro C, D'Anneo A, Carlisi D, Giuliano M, Gammazza AM, Barone R, Longhitano L, Cappello F, Emanuele S and Distefano A: Ethanol-mediated stress promotes autophagic survival and aggressiveness of colon cancer cells via activation of Nrf2/HO-1 pathway. Cancers (Basel). 11(505)2019.PubMed/NCBI View Article : Google Scholar
51	Schaaf MB, Keulers TG, Vooijs MA and Rouschop KM: LC3/GABARAP family proteins: autophagy-(un)related functions. FASEB J. 30:3961–3978. 2016.PubMed/NCBI View Article : Google Scholar
52	Vera-Ramirez L, Vodnala SK, Nini R, Hunter KW and Green JE: Autophagy promotes the survival of dormant breast cancer cells and metastatic tumour recurrence. Nat Commun. 9(1944)2018.PubMed/NCBI View Article : Google Scholar
53	Ma J, Dong C and Ji C: MicroRNA and drug resistance. Cancer Gene Ther. 17:523–531. 2010.PubMed/NCBI View Article : Google Scholar