Autophagy inhibition and microRNA‑199a‑5p upregulation in paclitaxel‑resistant A549/T lung cancer cells

Zeng,Tianjiao; Xu,Mingshi; Zhang,Wanli; Gu,Xiaofan; Zhao,Fangqing; Liu,Xuan; Zhang,Xiongwen

doi:10.3892/or.2021.8100

Autophagy inhibition and microRNA‑199a‑5p upregulation in paclitaxel‑resistant A549/T lung cancer cells

Authors:
- Tianjiao Zeng
- Mingshi Xu
- Wanli Zhang
- Xiaofan Gu
- Fangqing Zhao
- Xuan Liu
- Xiongwen Zhang
View Affiliations

Affiliations: Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200062, P.R. China, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China
Published online on: June 1, 2021 https://doi.org/10.3892/or.2021.8100
Article Number: 149
Copyright: © Zeng 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

Multidrug resistance (MDR) is one of the major reasons for the clinical failure of cancer chemotherapy. Autophagy activation serves a crucial role in MDR. However, the specific molecular mechanism linking autophagy with MDR remains unknown. The results of the present study demonstrated that autophagy was inhibited and microRNA (miR)‑199a‑5p levels were upregulated in MDR model lung cancer cells (A549/T and H1299/T) compared with those in the parental cell lines. Paclitaxel (PTX) treatment increased the expression levels of miR‑199a‑5p in parental lung cancer cells compared with those in PTX‑untreated cells, and these expression levels were negatively correlated with PTX sensitivity of the cells. miR‑199a‑5p knockdown in A549/T cells induced autophagy and resensitized cells to multiple chemotherapeutic drugs including PTX, taxotere, topotecan, SN38, oxaliplatin and vinorelbine. By contrast, miR‑199a‑5p overexpression in A549 cells suppressed autophagy and desensitized cells to these chemotherapeutic drugs. Mechanistically, the results of the present study demonstrated that miR‑199a‑5p blocked autophagy by activating the PI3K/Akt/mTOR signaling pathway and inhibiting the protein expression of autophagy‑related 5. Furthermore, p62 protein was identified as a direct target of miR‑199a‑5p; miR‑199a‑5p bound to p62 mRNA to decrease its mRNA and protein expression levels. In conclusion, the results of the present study suggested that miR‑199a‑5p may contribute to MDR development in lung cancer cells by inhibiting autophagy and targeting p62. The regulatory effect of miR‑199a‑5p on autophagy may provide novel insights for future multidrug‑resistant lung cancer chemotherapy.

View Figures

View References

1	Alfarouk KO, Stock CM, Taylor S, Walsh M, Muddathir AK, Verduzco D, Bashir AH, Mohammed OY, Elhassan GO, Harguindey S, et al: Resistance to cancer chemotherapy: Failure in drug response from ADME to P-gp. Cancer Cell Int. 15:712015. View Article : Google Scholar : PubMed/NCBI
2	Wu Q, Yang Z, Nie Y, Shi Y and Fan D: Multi-drug resistance in cancer chemotherapeutics: Mechanisms and lab approaches. Cancer Lett. 347:159–166. 2014. View Article : Google Scholar : PubMed/NCBI
3	Mizushima N, Levine B, Cuervo AM and Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 451:1069–1075. 2008. View Article : Google Scholar : PubMed/NCBI
4	Feng Y, He D, Yao Z and Klionsky DJ: The machinery of macroautophagy. Cell Res. 24:24–41. 2014. View Article : Google Scholar : PubMed/NCBI
5	Tanida I, Ueno T and Kominami E: LC3 and autophagy. Methods Mol Biol. 445:77–88. 2008. View Article : Google Scholar : PubMed/NCBI
6	Runwal G, Stamatakou E, Siddiqi FH, Puri C, Zhu Y and Rubinsztein DC: LC3-positive structures are prominent in autophagy-deficient cells. Sci Rep. 9:101472019. View Article : Google Scholar : PubMed/NCBI
7	Oh S, Xiaofei E, Ni D, Pirooz SD, Lee JY, Lee D, Zhao Z, Lee S, Lee H, Ku B, et al: Downregulation of autophagy by Bcl-2 promotes MCF7 breast cancer cell growth independent of its inhibition of apoptosis. Cell Death Differ. 18:452–464. 2011. View Article : Google Scholar : PubMed/NCBI
8	Yang J, Pi C and Wang G: Inhibition of PI3K/Akt/mTOR pathway by apigenin induces apoptosis and autophagy in hepatocellular carcinoma cells. Biomed Pharmacother. 103:699–707. 2018. View Article : Google Scholar : PubMed/NCBI
9	Tekirdag KA, Korkmaz G, Ozturk DG, Agami R and Gozuacik D: MIR181A regulates starvation- and rapamycin-induced autophagy through targeting of ATG5. Autophagy. 9:374–385. 2013. View Article : Google Scholar : PubMed/NCBI
10	Fang LM, Li B, Guan JJ, Xu HD, Shen GH, Gao QG and Qin ZH: Transcription factor EB is involved in autophagy-mediated chemoresistance to doxorubicin in human cancer cells. Acta Pharmacol Sin. 38:1305–1316. 2017. View Article : Google Scholar : PubMed/NCBI
11	Mishima Y, Terui Y, Mishima Y, Taniyama A, Kuniyoshi R, Takizawa T, Kimura S, Ozawa K and Hatake K: Autophagy and autophagic cell death are next targets for elimination of the resistance to tyrosine kinase inhibitors. Cancer Sci. 99:2200–2208. 2008. View Article : Google Scholar : PubMed/NCBI
12	Hu YL, Jahangiri A, Delay M and Aghi MK: Tumor cell autophagy as an adaptive response mediating resistance to treatments such as antiangiogenic therapy. Cancer Res. 72:4294–4299. 2012. View Article : Google Scholar : PubMed/NCBI
13	Sun WL, Chen J, Wang YP and Zheng H: Autophagy protects breast cancer cells from epirubicin-induced apoptosis and facilitates epirubicin-resistance development. Autophagy. 7:1035–1044. 2011. View Article : Google Scholar : PubMed/NCBI
14	Zhang P, Liu X, Li H, Chen Z, Yao X, Jin J and Ma X: TRPC5-induced autophagy promotes drug resistance in breast carcinoma via CaMKKβ/AMPKα/mTOR pathway. Sci Rep. 7:31582017. View Article : Google Scholar : PubMed/NCBI
15	Kanzawa T, Germano IM, Komata T, Ito H, Kondo Y and Kondo S: Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ. 11:448–457. 2004. View Article : Google Scholar : PubMed/NCBI
16	Lv XA, Wang B, Xu XH, Lei P, Bin W, Dong XX, Zeng CH and Du QW: Curcumin re-sensitizes multidrug resistant (MDR) breast cancer to cisplatin through inducing autophagy by decreasing CCAT1 expression. RSC Adv. 7:33572–33579. 2017. View Article : Google Scholar
17	Wu J, Li W, Ning J, Yu W, Rao T and Cheng F: Long noncoding RNA UCA1 targets miR-582-5p and contributes to the progression and drug resistance of bladder cancer cells through ATG7-mediated autophagy inhibition. Onco Targets Ther. 12:495–508. 2019. View Article : Google Scholar : PubMed/NCBI
18	Chen S, Wu J, Jiao K, Wu Q, Ma J, Chen D, Kang J and Zhao G, Shi Y, Fan D and Zhao G: MicroRNA-495-3p inhibits multidrug resistance by modulating autophagy through GRP78/mTOR axis in gastric cancer. Cell Death Dis. 9:10702018. View Article : Google Scholar : PubMed/NCBI
19	Li Y, Jiang W, Hu Y, Da Z, Zeng C, Tu M, Deng Z and Xiao W: MicroRNA-199a-5p inhibits cisplatin-induced drug resistance via inhibition of autophagy in osteosarcoma cells. Oncol Lett. 12:4203–4208. 2016. View Article : Google Scholar : PubMed/NCBI
20	Li Y, Zhang G, Wu B, Yang W and Liu Z: MiR-199a-5p represses protective autophagy and overcomes chemoresistance by directly targeting DRAM1 in acute myeloid leukemia. J Oncol. 2019:56134172019. View Article : Google Scholar : PubMed/NCBI
21	Chen PH, Liu AJ, Ho KH, Chiu YT, Anne Lin ZH, Lee YT, Shih CM and Chen KC: MicroRNA-199a/b-5p enhance imatinib efficacy via repressing WNT2 signaling-mediated protective autophagy in imatinib-resistant chronic myeloid leukemia cells. Chem Biol Interact. 291:144–151. 2018. View Article : Google Scholar : PubMed/NCBI
22	Li Y, Li L, Guan Y, Liu X, Meng Q and Guo Q: MiR-92b regulates the cell growth, cisplatin chemosensitivity of A549 non small cell lung cancer cell line and target PTEN. Biochem Biophys Res Commun. 440:604–610. 2013. View Article : Google Scholar : PubMed/NCBI
23	Li Y, Chen L, Feng L, Zhu M, Shen Q, Fang Y, Liu X and Zhang X: NEK2 promotes proliferation, migration and tumor growth of gastric cancer cells via regulating KDM5B/H3K4me3. Am J Cancer Res. 9:2364–2378. 2019.PubMed/NCBI
24	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. View Article : Google Scholar : PubMed/NCBI
25	Mauthe M, Orhon I, Rocchi C, Zhou X, Luhr M, Hijlkema KJ, Coppes RP, Engedal N, Mari M and Reggiori F: Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion. Autophagy. 14:1435–1455. 2018. View Article : Google Scholar : PubMed/NCBI
26	Moore CE, Wang X, Xie J, Pickford J, Barron J, Regufe da Mota S, Versele M and Proud CG: Elongation factor 2 kinase promotes cell survival by inhibiting protein synthesis without inducing autophagy. Cell Signal. 28:284–293. 2016. View Article : Google Scholar : PubMed/NCBI
27	Chen XL, Liu P, Zhu WL and Lou LG: DCZ5248, a novel dual inhibitor of Hsp90 and autophagy, exerts antitumor activity against colon cancer. Acta Pharmacol Sin. 42:132–141. 2021. View Article : Google Scholar : PubMed/NCBI
28	Duprez L, Wirawan E, Vanden Berghe T and Vandenabeele P: Major cell death pathways at a glance. Microbes Infect. 11:1050–1062. 2009. View Article : Google Scholar : PubMed/NCBI
29	Chang H and Zou Z: Targeting autophagy to overcome drug resistance: Further developments. J Hematol Oncol. 13:1592020. View Article : Google Scholar : PubMed/NCBI
30	Yoon JH, Ahn SG, Lee BH, Jung SH and Oh SH: Role of autophagy in chemoresistance: Regulation of the ATM-mediated DNA-damage signaling pathway through activation of DNA-PKcs and PARP-1. Biochem Pharmacol. 83:747–757. 2012. View Article : Google Scholar : PubMed/NCBI
31	Li S and Wei Y: Association of HMGB1, BRCA1 and P62 expression in ovarian cancer and chemotherapy sensitivity. Oncol Lett. 15:9572–9576. 2018.PubMed/NCBI
32	Huang CY, Chiang SF, Chen WT, Ke TW, Chen TW, You YS, Lin CY, Chao KSC and Huang CY: HMGB1 promotes ERK-mediated mitochondrial Drp1 phosphorylation for chemoresistance through RAGE in colorectal cancer. Cell Death Dis. 9:10042018. View Article : Google Scholar : PubMed/NCBI
33	Zheng H, Chen JN, Yu X, Jiang P, Yuan L, Shen HS, Zhao LH, Chen PF and Yang M: HMGB1 enhances drug resistance and promotes in vivo tumor growth of lung cancer cells. DNA Cell Biol. 35:622–627. 2016. View Article : Google Scholar : PubMed/NCBI
34	Wirawan E, Vanden Berghe T, Lippens S, Agostinis P and Vandenabeele P: Autophagy: For better or for worse. Cell Res. 22:43–61. 2012. View Article : Google Scholar : PubMed/NCBI
35	Xie T, Li SJ, Guo MR, Wu Y, Wang HY, Zhang K, Zhang X, Ouyang L and Liu J: Untangling knots between autophagic targets and candidate drugs, in cancer therapy. Cell Prolif. 48:119–139. 2015. View Article : Google Scholar : PubMed/NCBI
36	Wu H, Yang JM, Jin S, Zhang H and Hait WN: Elongation factor-2 kinase regulates autophagy in human glioblastoma cells. Cancer Res. 66:3015–3023. 2006. View Article : Google Scholar : PubMed/NCBI
37	Pan Z, Chen Y, Liu J, Jiang Q, Yang S, Guo L and He G: Design, synthesis, and biological evaluation of polo-like kinase 1/eukaryotic elongation factor 2 kinase (PLK1/EEF2K) dual inhibitors for regulating breast cancer cells apoptosis and autophagy. Eur J Med Chem. 144:517–528. 2018. View Article : Google Scholar : PubMed/NCBI
38	Xie CM, Liu XY, Sham KW, Lai JM and Cheng CH: Silencing of EEF2K (eukaryotic elongation factor-2 kinase) reveals AMPK-ULK1-dependent autophagy in colon cancer cells. Autophagy. 10:1495–1508. 2014. View Article : Google Scholar : PubMed/NCBI
39	Sun D, Zhu L, Zhao Y, Jiang Y, Chen L, Yu Y and Ouyang L: Fluoxetine induces autophagic cell death via eEF2K-AMPK-mTOR-ULK complex axis in triple negative breast cancer. Cell Prolif. 51:e124022018. View Article : Google Scholar : PubMed/NCBI
40	Wang H, Gao H, Duan S and Song X: Inhibition of microRNA-199a-5p reduces the replication of HCV via regulating the pro-survival pathway. Virus Res. 208:7–12. 2015. View Article : Google Scholar : PubMed/NCBI
41	Wang G, Sun D, Li W and Xin Y: AKNA is a potential prognostic biomarker in gastric cancer and function as a tumor suppressor by modulating EMT-related pathways. Biomed Res Int. 2020:67267592020.PubMed/NCBI
42	Murugesan M and Premkumar K: Integrative miRNA-mRNA functional analysis identifies miR-182 as a potential prognostic biomarker in breast cancer. Mol Omics. Apr 22–2021.(Epub ahead of print). doi: 10.1039/d0mo00160k. View Article : Google Scholar : PubMed/NCBI
43	Zhang K, Zhang X, Cai Z, Zhou J, Cao R, Zhao Y, Chen Z, Wang D, Ruan W, Zhao Q, et al: A novel class of microRNA-recognition elements that function only within open reading frames. Nat Struct Mol Biol. 25:1019–1027. 2018. View Article : Google Scholar : PubMed/NCBI
44	Moscat J, Karin M and Diaz-Meco MT: p62 in cancer: Signaling adaptor beyond autophagy. Cell. 167:606–609. 2016. View Article : Google Scholar : PubMed/NCBI
45	Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, Stenmark H and Johansen T: p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol. 171:603–614. 2005. View Article : Google Scholar : PubMed/NCBI
46	Peng H, Yang J, Li G, You Q, Han W, Li T, Gao D, Xie X, Lee BH, Du J, et al: Ubiquitylation of p62/sequestosome1 activates its autophagy receptor function and controls selective autophagy upon ubiquitin stress. Cell Res. 27:657–674. 2017. View Article : Google Scholar : PubMed/NCBI
47	Gao Z, Gammoh N, Wong PM, Erdjument-Bromage H, Tempst P and Jiang X: Processing of autophagic protein LC3 by the 20S proteasome. Autophagy. 6:126–137. 2010. View Article : Google Scholar : PubMed/NCBI
48	Bjørkøy G, Lamark T, Pankiv S, Øvervatn A, Brech A and Johansen T: Monitoring autophagic degradation of p62/SQSTM1. Methods Enzymol. 452:181–197. 2009. View Article : Google Scholar
49	Zhang CF, Gruber F, Ni C, Mildner M, Koenig U, Karner S, Barresi C, Rossiter H, Narzt MS, Nagelreiter IM, et al: Suppression of autophagy dysregulates the antioxidant response and causes premature senescence of melanocytes. J Invest Dermatol. 135:1348–1357. 2015. View Article : Google Scholar : PubMed/NCBI