Top

Journal of Experimental & Clinical Cancer Research

Published in:

01-12-2021 | Ovarian Cancer | Research

ALKBH5-HOXA10 loop-mediated JAK2 m6A demethylation and cisplatin resistance in epithelial ovarian cancer

Authors: Sipei Nie, Lin Zhang, Jinhui Liu, Yicong Wan, Yi Jiang, Jing Yang, Rui Sun, Xiaolling Ma, Guodong Sun, Huangyang Meng, Mengting Xu, Wenjun Cheng

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2021

Abstract

Background

Chemotherapy resistance remains a barrier to improving the prognosis of epithelial ovarian cancer (EOC). ALKBH5 has recently been shown to be one of the RNA N6-methyladenosine (m6A) demethyltransferases associated with various cancers, but its role in cancer therapeutic resistance remains unclear. This study aimed to investigate the role of AlkB homolog 5 (ALKBH5) in cisplatin-resistant EOC.

Methods

Functional assays were performed both in vitro and in vivo. RNA sequencing (RNA-seq), m6A-modified RNA immunoprecipitation sequencing (MeRIP-seq), chromatin immunoprecipitation, RNA immunoprecipitation, and luciferase reporter and actinomycin-D assays were performed to investigate RNA/RNA interaction and m6A modification of the ALKBH5-HOXA10 loop.

Results

ALKBH5 was upregulated in cisplatin-resistant EOC and promoted cancer cell cisplatin resistance both in vivo and in vitro. Notably, HOXA10 formed a loop with ALKBH5 and was found to be the upstream transcription factor of ALKBH5. HOXA10 overexpression also facilitated EOC cell chemoresistance both in vivo and in vitro. Collective results of MeRIP-seq and RNA-seq showed that JAK2 is the m6A-modified gene targeted by ALKBH5. The JAK2/STAT3 signaling pathway was activated by overexpression of the ALKBH5-HOXA10 loop, resulting in EOC chemoresistance. Cell sensitivity to cisplatin was rescued by ALKBH5 and HOXA10 knockdown or inhibition of the JAK2/STAT3 signaling pathway in EOC cells overexpressing ALKBH5-HOXA10.

Conclusions

The ALKBH5-HOXA10 loop jointly activates the JAK2/STAT3 signaling pathway by mediating JAK2 m6A demethylation, promoting EOC resistance to cisplatin. Thus, inhibition of the expression of the ALKBH5-HOXA10 loop may be a potential strategy to overcome cisplatin resistance in EOC.

Available only for authorised users

Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70(1):7–30. https://doi.org/10.3322/caac.21590.CrossRefPubMed

Stewart C, Ralyea C, Lockwood S. Ovarian Cancer: An integrated review. Semin Oncol Nurs. 2019;35(2):151–6. https://doi.org/10.1016/j.soncn.2019.02.001.CrossRefPubMed

Shen H, Wang GC, Li X, Ge X, Wang M, Shi ZM, et al. S6K1 blockade overcomes acquired resistance to EGFR-TKIs in non-small cell lung cancer. Oncogene. 2020;39(49):7181–95. https://doi.org/10.1038/s41388-020-01497-4.CrossRefPubMedPubMedCentral

Thompson PA, Eam B, Young NP, Fish S, Chen J, Barrera M, et al. Targeting oncogene mRNA translation in B cell malignancies with eFT226, a potent and selective inhibitor of eIF4A1. Mol Cancer Ther. 2020.

Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, et al. Molecular mechanisms of cisplatin resistance. Oncogene. 2012;31(15):1869–83. https://doi.org/10.1038/onc.2011.384.CrossRefPubMed

Liang Z, Kidwell RL, Deng H, Xie Q. Epigenetic N6-methyladenosine modification of RNA and DNA regulates cancer. Cancer Biol Med. 2020;17(1):9–19. https://doi.org/10.20892/j.issn.2095-3941.2019.0347.CrossRefPubMedPubMedCentral

Wang X, Huang J, Zou T, Yin P. Human m(6) a writers: two subunits, 2 roles. RNA Biol. 2017;14(3):300–4. https://doi.org/10.1080/15476286.2017.1282025.CrossRefPubMedPubMedCentral

Shi H, Wei J, He C. Where, when, and how: context-dependent functions of RNA methylation writers, readers, and erasers. Mol Cell. 2019;74(4):640–50. https://doi.org/10.1016/j.molcel.2019.04.025.CrossRefPubMedPubMedCentral

Xu J, Wan Z, Tang M, Lin Z, Jiang S, Ji L, et al. N(6)-methyladenosine-modified CircRNA-SORE sustains sorafenib resistance in hepatocellular carcinoma by regulating beta-catenin signaling. Mol Cancer. 2020;19(1):163. https://doi.org/10.1186/s12943-020-01281-8.CrossRefPubMedPubMedCentral

10.

Liu X, Gonzalez G, Dai X, Miao W, Yuan J, Huang M, et al. Adenylate kinase 4 modulates the resistance of breast Cancer cells to tamoxifen through an m(6)A-based Epitranscriptomic mechanism. Mol Ther. 2020;28(12):2593–604. https://doi.org/10.1016/j.ymthe.2020.09.007.CrossRefPubMedPubMedCentral

11.

Jin D, Guo J, Wu Y, Du J, Yang L, Wang X, et al. m(6) a mRNA methylation initiated by METTL3 directly promotes YAP translation and increases YAP activity by regulating the MALAT1-miR-1914-3p-YAP axis to induce NSCLC drug resistance and metastasis. J Hematol Oncol. 2019;12(1):135. https://doi.org/10.1186/s13045-019-0830-6.CrossRefPubMedPubMedCentral

12.

Yu H, Yang X, Tang J, Si S, Zhou Z, Lu J, et al. ALKBH5 inhibited cell proliferation and sensitized bladder Cancer cells to cisplatin by m6A-CK2alpha-mediated glycolysis. Mol Ther Nucleic Acids. 2021;23:27–41. https://doi.org/10.1016/j.omtn.2020.10.031.CrossRefPubMed

13.

Zhu H, Gan X, Jiang X, Diao S, Wu H, Hu J. ALKBH5 inhibited autophagy of epithelial ovarian cancer through miR-7 and BCL-2. J Exp Clin Cancer Res. 2019;38(1):163. https://doi.org/10.1186/s13046-019-1159-2.CrossRefPubMedPubMedCentral

14.

Fukumoto T, Zhu H, Nacarelli T, Karakashev S, Fatkhutdinov N, Wu S, et al. N(6)-methylation of adenosine of FZD10 mRNA contributes to PARP inhibitor resistance. Cancer Res. 2019;79(11):2812–20. https://doi.org/10.1158/0008-5472.CAN-18-3592.CrossRefPubMedPubMedCentral

15.

Armstrong DK, Alvarez RD, Bakkum-Gamez JN, Barroilhet L, Behbakht K, Berchuck A, et al. NCCN guidelines insights: ovarian Cancer, version 1.2019. J Natl Compr Cancer Netw. 2019;17(8):896–909. https://doi.org/10.6004/jnccn.2019.0039.CrossRef

16.

Gong M, Luo C, Meng H, Li S, Nie S, Jiang Y, et al. Upregulated LINC00565 accelerates ovarian Cancer progression by targeting GAS6. Onco Targets Ther. 2019;12:10011–22. https://doi.org/10.2147/OTT.S227758.CrossRefPubMedPubMedCentral

17.

Turinetto V, Giachino C. Multiple facets of histone variant H2AX: a DNA double-strand-break marker with several biological functions. Nucleic Acids Res. 2015;43(5):2489–98. https://doi.org/10.1093/nar/gkv061.CrossRefPubMedPubMedCentral

18.

Cheng W, Liu J, Yoshida H, Rosen D, Naora H. Lineage infidelity of epithelial ovarian cancers is controlled by HOX genes that specify regional identity in the reproductive tract. Nat Med. 2005;11(5):531–7. https://doi.org/10.1038/nm1230.CrossRefPubMed

19.

Tang W, Jiang Y, Mu X, Xu L, Cheng W, Wang X. MiR-135a functions as a tumor suppressor in epithelial ovarian cancer and regulates HOXA10 expression. Cell Signal. 2014;26(7):1420–6. https://doi.org/10.1016/j.cellsig.2014.03.002.CrossRefPubMed

20.

Liu J, Jiang Y, Wan Y, Zhou S, Thapa S, Cheng W. MicroRNA665 suppresses the growth and migration of ovarian cancer cells by targeting HOXA10. Mol Med Rep. 2018;18(3):2661–8. https://doi.org/10.3892/mmr.2018.9252.CrossRefPubMedPubMedCentral

21.

Buchert M, Burns CJ, Ernst M. Targeting JAK kinase in solid tumors: emerging opportunities and challenges. Oncogene. 2016;35(8):939–51. https://doi.org/10.1038/onc.2015.150.CrossRefPubMed

22.

Liu J, Harada BT, He C. Regulation of gene expression by N(6)-methyladenosine in Cancer. Trends Cell Biol. 2019;29(6):487–99. https://doi.org/10.1016/j.tcb.2019.02.008.CrossRefPubMedPubMedCentral

23.

Hua W, Zhao Y, Jin X, Yu D, He J, Xie D, et al. METTL3 promotes ovarian carcinoma growth and invasion through the regulation of AXL translation and epithelial to mesenchymal transition. Gynecol Oncol. 2018;151(2):356–65. https://doi.org/10.1016/j.ygyno.2018.09.015.CrossRefPubMed

24.

Huang H, Wang Y, Kandpal M, Zhao G, Cardenas H, Ji Y, et al. FTO-dependent N (6)-Methyladenosine modifications inhibit ovarian Cancer stem cell self-renewal by blocking cAMP signaling. Cancer Res. 2020;80(16):3200–14. https://doi.org/10.1158/0008-5472.CAN-19-4044.CrossRefPubMedPubMedCentral

25.

Liu T, Wei Q, Jin J, Luo Q, Liu Y, Yang Y, et al. The m6A reader YTHDF1 promotes ovarian cancer progression via augmenting EIF3C translation. Nucleic Acids Res. 2020;48(7):3816–31. https://doi.org/10.1093/nar/gkaa048.CrossRefPubMedPubMedCentral

26.

Muller S, Glass M, Singh AK, Haase J, Bley N, Fuchs T, et al. IGF2BP1 promotes SRF-dependent transcription in cancer in a m6A- and miRNA-dependent manner. Nucleic Acids Res. 2019;47(1):375–90. https://doi.org/10.1093/nar/gky1012.CrossRefPubMed

27.

Guo T, Liu DF, Peng SH, Xu AM. ALKBH5 promotes colon cancer progression by decreasing methylation of the lncRNA NEAT1. Am J Transl Res. 2020;12(8):4542–9.PubMedPubMedCentral

28.

Pu X, Gu Z, Gu Z. ALKBH5 regulates IGF1R expression to promote the proliferation and Tumorigenicity of endometrial Cancer. J Cancer. 2020;11(19):5612–22. https://doi.org/10.7150/jca.46097.CrossRefPubMedPubMedCentral

29.

Zhang X, Wang F, Wang Z, Yang X, Yu H, Si S, et al. ALKBH5 promotes the proliferation of renal cell carcinoma by regulating AURKB expression in an m(6)A-dependent manner. Ann Transl Med. 2020;8(10):646. https://doi.org/10.21037/atm-20-3079.CrossRefPubMedPubMedCentral

30.

Guo X, Li K, Jiang W, Hu Y, Xiao W, Huang Y, et al. RNA demethylase ALKBH5 prevents pancreatic cancer progression by posttranscriptional activation of PER1 in an m6A-YTHDF2-dependent manner. Mol Cancer. 2020;19(1):91. https://doi.org/10.1186/s12943-020-01158-w.CrossRefPubMedPubMedCentral

31.

Chen Y, Zhao Y, Chen J, Peng C, Zhang Y, Tong R, et al. ALKBH5 suppresses malignancy of hepatocellular carcinoma via m(6)A-guided epigenetic inhibition of LYPD1. Mol Cancer. 2020;19(1):123. https://doi.org/10.1186/s12943-020-01239-w.CrossRefPubMedPubMedCentral

32.

Kowalski-Chauvel A, Lacore MG, Arnauduc F, Delmas C, Toulas C, Cohen-Jonathan-Moyal E, et al. The m6A RNA demethylase ALKBH5 promotes Radioresistance and invasion capability of glioma stem cells. Cancers (Basel). 2020;13(1). https://doi.org/10.3390/cancers13010040.

33.

Li N, Kang Y, Wang L, Huff S, Tang R, Hui H, et al. ALKBH5 regulates anti-PD-1 therapy response by modulating lactate and suppressive immune cell accumulation in tumor microenvironment. Proc Natl Acad Sci U S A. 2020;117(33):20159–70. https://doi.org/10.1073/pnas.1918986117.CrossRefPubMedPubMedCentral

34.

Daftary GS, Taylor HS. Endocrine regulation of HOX genes. Endocr Rev. 2006;27(4):331–55. https://doi.org/10.1210/er.2005-0018.CrossRefPubMed

35.

Richie JP. Re: loss of nuclear functions of HOXA10 is associated with testicular Cancer proliferation. J Urol. 2019;202(2):213. https://doi.org/10.1097/JU.0000000000000301.CrossRefPubMed

36.

Trissal MC, Wong TN, Yao JC, Ramaswamy R, Kuo I, Baty J, et al. MIR142 loss-of-function mutations Derepress ASH1L to increase HOXA gene expression and promote Leukemogenesis. Cancer Res. 2018;78(13):3510–21. https://doi.org/10.1158/0008-5472.CAN-17-3592.CrossRefPubMedPubMedCentral

37.

Yoshida H, Broaddus R, Cheng W, Xie S, Naora H. Deregulation of the HOXA10 homeobox gene in endometrial carcinoma: role in epithelial-mesenchymal transition. Cancer Res. 2006;66(2):889–97. https://doi.org/10.1158/0008-5472.CAN-05-2828.CrossRefPubMed

38.

Liu J, Li C, Jiang Y, Wan Y, Zhou S, Cheng W. Tumor-suppressor role of miR-139-5p in endometrial cancer. Cancer Cell Int. 2018;18(1):51. https://doi.org/10.1186/s12935-018-0545-8.CrossRefPubMedPubMedCentral

39.

Yu H, Pardoll D, Jove R. STATs in cancer inflammation and immunity: a leading role for STAT3. Nat Rev Cancer. 2009;9(11):798–809. https://doi.org/10.1038/nrc2734.CrossRefPubMedPubMedCentral

40.

Yogev O, Almeida GS, Barker KT, George SL, Kwok C, Campbell J, et al. In vivo modeling of Chemoresistant neuroblastoma provides new insights into Chemorefractory disease and metastasis. Cancer Res. 2019;79(20):5382–93. https://doi.org/10.1158/0008-5472.CAN-18-2759.CrossRefPubMed

41.

Raghavan S, Mehta P, Ward MR, Bregenzer ME, Fleck E, Tan L, et al. Personalized medicine-based approach to model patterns of Chemoresistance and tumor recurrence using ovarian Cancer stem cell spheroids. Clin Cancer Res. 2017;23(22):6934–45. https://doi.org/10.1158/1078-0432.CCR-17-0133.CrossRefPubMedPubMedCentral

42.

Zhang Z, Wang F, Du C, Guo H, Ma L, Liu X, et al. BRM/SMARCA2 promotes the proliferation and chemoresistance of pancreatic cancer cells by targeting JAK2/STAT3 signaling. Cancer Lett. 2017;402:213–24. https://doi.org/10.1016/j.canlet.2017.05.006.CrossRefPubMed

43.

Wu R, Liu Y, Zhao Y, Bi Z, Yao Y, Liu Q, et al. m(6) a methylation controls pluripotency of porcine induced pluripotent stem cells by targeting SOCS3/JAK2/STAT3 pathway in a YTHDF1/YTHDF2-orchestrated manner. Cell Death Dis. 2019;10(3):171. https://doi.org/10.1038/s41419-019-1417-4.CrossRefPubMedPubMedCentral

44.

Wang X, Lu Z, Gomez A, Hon GC, Yue Y, Han D, et al. N6-methyladenosine-dependent regulation of messenger RNA stability. Nature. 2014;505(7481):117–20. https://doi.org/10.1038/nature12730.CrossRefPubMed

Title: ALKBH5-HOXA10 loop-mediated JAK2 m6A demethylation and cisplatin resistance in epithelial ovarian cancer
Authors: Sipei Nie
Lin Zhang
Jinhui Liu
Yicong Wan
Yi Jiang
Jing Yang
Rui Sun
Xiaolling Ma
Guodong Sun
Huangyang Meng
Mengting Xu
Wenjun Cheng
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Ovarian Cancer
Ovarian Cancer
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2021
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-021-02088-1

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2021

Differences between intrinsic and acquired nucleoside analogue resistance in acute myeloid leukaemia cells

Claudin-2 inhibits renal clear cell carcinoma progression by inhibiting YAP-activation

PARP inhibitors in gastric cancer: beacon of hope

Lnc-GAN1 expression is associated with good survival and suppresses tumor progression by sponging mir-26a-5p to activate PTEN signaling in non-small cell lung cancer

A novel humanized Frizzled-7-targeting antibody enhances antitumor effects of Bevacizumab against triple-negative breast cancer via blocking Wnt/β-catenin signaling pathway

The CD112R/CD112 axis: a breakthrough in cancer immunotherapy

Keynote webinar | Spotlight on antibody–drug conjugates in cancer