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Knockdown of long non-coding HOTAIR enhances the sensitivity to progesterone in endometrial cancer by epigenetic regulation of progesterone receptor isoform B

  • Shuqi Chi
  • Yan Liu
  • Xing Zhou
  • Dilu Feng
  • Xianjin Xiao
  • Wenliang Li
  • Yingchao Zhao
  • Hongbo Wang
Original Article
  • 88 Downloads

Abstract

Purpose

Progesterone, particularly medroxyprogesterone acetate (MPA) has been mainly used for young endometrial carcinoma (EC) patients with conservative treatment. However, its treatment benefits are limited by insensitivity or acquired resistance. In this study, we aim to investigate the effect of long non-coding RNA HOTAIR on progesterone sensitivity in endometrial cancer, as well as the underlying mechanisms.

Methods

The expression of HOTAIR was detected by quantitative real-time PCR. The impact of MPA on the endometrial cancer cells was examined by MTT, colony formation, apoptosis-related protein detection and flow cytometry. Chromatin immunoprecipitation (ChIP) assay was performed to detect the regulatory mechanism between HOTAIR and progesterone receptor B (PRB). We further confirm the function of HOTAIR in vivo though xenograft tumor assay.

Results

We found that HOTAIR inversely correlated with PRB expression in endometrial carcinoma. Knockdown of HOTAIR promoted the MPA sensitivity by upregulating PRB, which can be largely reversed by PRB downregulation. Moreover, inhibiting LSD1, a HOTAIR-associated protein that removed activating H3K4me2 chromatin marks, induced PRB expression and promoted apoptosis induced by MPA. We further showed that silencing HOTAIR strengthened the H3K4me2 occupation on the promotor of PRB.

Conclusions

Our findings provide compelling evidence that HOTAIR and LSD1 collaboratively repress PRB expression and thus mediate progesterone sensitivity in endometrial carcinoma cells. HOTAIR is a potential predictor for progesterone response in EC and down-regulated expression of HOTAIR might be an effective strategy for overcoming progesterone resistance.

Keywords

Endometrial cancer HOTAIR Progesterone sensitivity Epigenetic modification 

Notes

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants and animals were in accordance with the ethical standards of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

280_2018_3727_MOESM1_ESM.docx (396 kb)
Supplementary material 1 (DOCX 395 KB)

References

  1. 1.
    Ferlay J, Steliarova-Foucher E, Lortet-Tieulent J, Rosso S, Coebergh JW, Comber H, Forman D, Bray F (2013) Cancer incidence and mortality patterns in Europe: estimates for 40 countries in 2012. Eur J Cancer 49(6):1374–1403CrossRefGoogle Scholar
  2. 2.
    Schindler AE (2009) Progestogen deficiency and endometrial cancer risk. Maturitas 62(4):334–337CrossRefGoogle Scholar
  3. 3.
    Kim JJ, Kurita T, Bulun SE (2013) Progesterone action in endometrial cancer, endometriosis, uterine fibroids, and breast cancer. Endocr Rev 34(1):130–162CrossRefGoogle Scholar
  4. 4.
    Erkanli S, Ayhan A (2010) Fertility-sparing therapy in young women with endometrial cancer: 2010 update. Int J Gynecol Cancer 20(7):1170–1187CrossRefGoogle Scholar
  5. 5.
    Kim JJ, Chapman-Davis E (2010) Role of progesterone in endometrial cancer. Semin Reprod Med 28(1):81–90CrossRefGoogle Scholar
  6. 6.
    Hahn HS, Yoon SG, Hong JS, Hong SR, Park SJ, Lim JY, Kwon YS, Lee IH, Lim KT, Lee KH, Shim JU, Mok JE, Kim TJ (2009) Conservative treatment with progestin and pregnancy outcomes in endometrial cancer. Int J Gynecol Cancer 19(6):1068–1073CrossRefGoogle Scholar
  7. 7.
    Wang S, Pudney J, Song J, Mor G, Schwartz PE, Zheng W (2003) Mechanisms involved in the evolution of progestin resistance in human endometrial hyperplasia—precursor of endometrial cancer. Gynecol Oncol 88(2):108–117CrossRefGoogle Scholar
  8. 8.
    Kastner P, Krust A, Turcotte B, Stropp U, Tora L, Gronemeyer H, Chambon P (1990) Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. EMBO j 9(5):1603–1614CrossRefGoogle Scholar
  9. 9.
    Dai D, Wolf DM, Litman ES, White MJ, Leslie KK (2002) Progesterone inhibits human endometrial cancer cell growth and invasiveness: down-regulation of cellular adhesion molecules through progesterone B receptors. Cancer Res 62(3):881–886PubMedGoogle Scholar
  10. 10.
    Sakaguchi H, Fujimoto J, Hong BL, Nakagawa Y, Tamaya T (2004) Drastic decrease of progesterone receptor form B but not A mRNA reflects poor patient prognosis in endometrial cancers. Gynecol Oncol 93(2):394–399CrossRefGoogle Scholar
  11. 11.
    Pijnenborg JM, Romano A, Dam-de Veen GC, Dunselman GA, Fischer DC, Groothuis PG, Kieback DG (2005) Aberrations in the progesterone receptor gene and the risk of recurrent endometrial carcinoma. J Pathol 205(5):597–605CrossRefGoogle Scholar
  12. 12.
    Neubauer NL, Ward EC, Patel P, Lu Z, Lee I, Blok LJ, Hanifi-Moghaddam P, Schink J, Kim JJ (2011) Progesterone receptor-B induction of BIRC3 protects endometrial cancer cells from AP1-59-mediated apoptosis. Horm Cancer 2(3):170–181CrossRefGoogle Scholar
  13. 13.
    Ward EC, Hoekstra AV, Blok LJ, Hanifi-Moghaddam P, Lurain JR, Singh DK, Buttin BM, Schink JC, Kim JJ (2008) The regulation and function of the forkhead transcription factor, Forkhead box O1, is dependent on the progesterone receptor in endometrial carcinoma. Endocrinology 149(4):1942–1950CrossRefGoogle Scholar
  14. 14.
    Utsunomiya H, Suzuki T, Ito K, Moriya T, Konno R, Sato S, Yaegashi N, Okamura K, Sasano H (2003) The correlation between the response to progestogen treatment and the expression of progesterone receptor B and 17beta-hydroxysteroid dehydrogenase type 2 in human endometrial carcinoma. Clin Endocrinol (Oxf) 58(6):696–703CrossRefGoogle Scholar
  15. 15.
    Morlando M, Fatica A (2018) Alteration of epigenetic regulation by long noncoding RNAs in cancer. Int J Mol Sci 19(2):570CrossRefGoogle Scholar
  16. 16.
    Lin C, Yang L (2018) Long noncoding RNA in cancer: wiring signaling circuitry. Trends Cell Biol 28(4):287–301CrossRefGoogle Scholar
  17. 17.
    Arun G, Diermeier SD, Spector DL (2018) Therapeutic targeting of long non-coding RNAs in cancer. Trends Mol Med 24(3):257–277CrossRefGoogle Scholar
  18. 18.
    Zhang HY, Liang F, Zhang JW, Wang F, Wang L, Kang XG (2017) Effects of long noncoding RNA-ROR on tamoxifen resistance of breast cancer cells by regulating microRNA-205. Cancer Chemother Pharmacol 79(2):327–337CrossRefGoogle Scholar
  19. 19.
    Wang L, Wang F, Na L, Yu J, Huang L, Meng ZQ, Chen Z, Chen H, Ming LL, Hua YQ (2018) LncRNA AB209630 inhibits gemcitabine resistance cell proliferation by regulating PI3K/AKT signaling in pancreatic ductal adenocarcinoma. Cancer Biomark 22(1):169–174CrossRefGoogle Scholar
  20. 20.
    You QY, Tao H, Ling B (2014) Long noncoding RNA HOX transcript antisense intergenic RNA (HOTAIR) as a foe and novel potential therapeutic target for endometrial carcinoma. Int J Gynecol Cancer 24(9):1536CrossRefGoogle Scholar
  21. 21.
    Bhan A, Mandal SS (2015) LncRNA HOTAIR: a master regulator of chromatin dynamics and cancer. Biochim Biophys Acta 1856(1):151–164PubMedPubMedCentralGoogle Scholar
  22. 22.
    Liu L, Cui S, Wan T, Li X, Tian W, Zhang R, Luo L, Shi Y (2018) Long non-coding RNA HOTAIR acts as a competing endogenous RNA to promote glioma progression by sponging miR-126-5p. J Cell Physiol 233(9):6822–6831CrossRefGoogle Scholar
  23. 23.
    Xiao Z, Qu Z, Chen Z, Fang Z, Zhou K, Huang Z, Guo X, Zhang Y (2018) LncRNA HOTAIR is a prognostic biomarker for the proliferation and chemoresistance of colorectal cancer via MiR-203a-3p-mediated Wnt/ß-catenin signaling pathway. Cell Physiol Biochem 46(3):1275–1285CrossRefGoogle Scholar
  24. 24.
    Yang Y, Jiang C, Yang Y, Guo L, Huang J, Liu X, Wu C, Zou J (2018) Silencing of LncRNA-HOTAIR decreases drug resistance of non-small cell lung cancer cells by inactivating autophagy via suppressing the phosphorylation of ULK1. Biochem Biophys Res Commun 497(4):1003–1010CrossRefGoogle Scholar
  25. 25.
    Zhou JJ, Cheng D, He XY, Meng Z, Ye HL, Chen RF (2017) Knockdown of long non-coding RNA HOTAIR sensitizes hepatocellular carcinoma cell to cisplatin by suppressing the STAT3/ABCB1 signaling pathway. Oncol Lett 14(6):7986–7992PubMedPubMedCentralGoogle Scholar
  26. 26.
    Shang C, Guo Y, Zhang H, Xue YX (2016) Long noncoding RNA HOTAIR is a prognostic biomarker and inhibits chemosensitivity to doxorubicin in bladder transitional cell carcinoma. Cancer Chemother Pharmacol 77(3):507–513CrossRefGoogle Scholar
  27. 27.
    Chu Y, Wang Y, Zhang G, Chen H, Dowdy SC, Xiong Y, Liu F, Zhang R, Li J, Jiang SW (2014) Chromatin composition alterations and the critical role of MeCP2 for epigenetic silencing of progesterone receptor-B gene in endometrial cancers. CMLS 71(17):3393–3408CrossRefGoogle Scholar
  28. 28.
    Cai B, Song XQ, Cai JP, Zhang S (2014) HOTAIR: a cancer-related long non-coding RNA. Neoplasma 61(4):379–391CrossRefGoogle Scholar
  29. 29.
    Huang M, Chen C, Geng J, Han D, Wang T, Xie T, Wang L, Wang Y, Wang C, Lei Z, Chu X (2017) Targeting KDM1A attenuates Wnt/beta-catenin signaling pathway to eliminate sorafenib-resistant stem-like cells in hepatocellular carcinoma. Cancer Lett 398:12–21CrossRefGoogle Scholar
  30. 30.
    Woo CJ, Kingston RE (2007) HOTAIR lifts noncoding RNAs to new levels. Cell 129(7):1257–1259CrossRefGoogle Scholar
  31. 31.
    Xue X, Yang YA, Zhang A, Fong KW, Kim J, Song B, Li S, Zhao JC, Yu J (2016) LncRNA HOTAIR enhances ER signaling and confers tamoxifen resistance in breast cancer. Oncogene 35(21):2746–2755CrossRefGoogle Scholar
  32. 32.
    Zhang A, Zhao JC, Kim J, Fong KW, Yang YA, Chakravarti D, Mo YY, Yu J (2015) LncRNA HOTAIR enhances the androgen-receptor-mediated transcriptional program and drives castration-resistant prostate cancer. Cell Rep 13(1):209–221CrossRefGoogle Scholar
  33. 33.
    Nie L, Zhao YB, Pan JL, Lei Y, Liu M, Long Y, Zhang JH, Hu Y, Xu MQ, Yuan DZ, Yue LM (2017) Progesterone-induced miR-152 inhibits the proliferation of endometrial epithelial cells by downregulating WNT-1. Reprod Sci 24(10):1444–1453CrossRefGoogle Scholar
  34. 34.
    Lee II, Maniar K, Lydon JP, Kim JJ (2016) Akt regulates progesterone receptor B-dependent transcription and angiogenesis in endometrial cancer cells. Oncogene 35(39):5191–5201CrossRefGoogle Scholar
  35. 35.
    Yu Y, Zhang M, Wang N, Li Q, Yang J, Yan S, He X, Ji G, Miao L (2018) Epigenetic silencing of tumor suppressor gene CDKN1A by oncogenic long non-coding RNA SNHG1 in cholangiocarcinoma. Cell Death Dis 9(7):746CrossRefGoogle Scholar
  36. 36.
    Ma Z, Peng P, Zhou J, Hui B, Ji H, Wang J, Wang K (2018) Long non-coding RNA SH3PXD2A-AS1 promotes cell progression partly through epigenetic silencing P57 and KLF2 in colorectal cancer. Cell Physiol Biochem 46(6):2197–2214CrossRefGoogle Scholar
  37. 37.
    Shi Y, Lan F, Matson C, Mulligan P, Whetstine JR, Cole PA, Casero RA, Shi Y (2004) Histone demethylation mediated by the nuclear amine oxidase homolog LSD1. Cell 119(7):941–953CrossRefGoogle Scholar
  38. 38.
    Metzger E, Wissmann M, Yin N, Muller JM, Schneider R, Peters AH, Gunther T, Buettner R, Schule R (2005) LSD1 demethylates repressive histone marks to promote androgen-receptor-dependent transcription. Nature 437(7057):436–439CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Shuqi Chi
    • 1
  • Yan Liu
    • 1
  • Xing Zhou
    • 1
  • Dilu Feng
    • 1
  • Xianjin Xiao
    • 4
  • Wenliang Li
    • 3
  • Yingchao Zhao
    • 2
  • Hongbo Wang
    • 1
  1. 1.Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Cancer Center, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  3. 3.Texas Therapeutics Institute, Brown Foundation Institute of Molecular MedicineUniversity of Texas Health Science Center at HoustonHoustonUSA
  4. 4.Centre of Reproductive Medicine/Family Planning Research Institute, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina

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