Hypoxia‑induced lactate dehydrogenase A protects cells from apoptosis in endometriosis

Zheng,Jinyan; Dai,Yongdong; Lin,Xiang; Huang,Qianmeng; Shi,Libing; Jin,Xiaoying; Liu,Na; Zhou,Feng; Zhang,Songying

doi:10.3892/mmr.2021.12276

Hypoxia‑induced lactate dehydrogenase A protects cells from apoptosis in endometriosis

Authors:
- Jinyan Zheng
- Yongdong Dai
- Xiang Lin
- Qianmeng Huang
- Libing Shi
- Xiaoying Jin
- Na Liu
- Feng Zhou
- Songying Zhang
View Affiliations

Affiliations: Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, P.R. China
Published online on: July 6, 2021 https://doi.org/10.3892/mmr.2021.12276
Article Number: 637
Copyright: © Zheng 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

The pathological expression and function of lactate dehydrogenase A (LDHA), a key enzyme that converts pyruvate into lactic acid during glycolysis, remains unknown in endometriosis. In the present study, LDHA expression in tissue samples was determined by immunohistochemistry. To examine whether LDHA was induced by hypoxia, primary cultured endometrial stromal cells (ESCs) and glandular epithelial Ishikawa cells were exposed to 1% O₂ (hypoxia) or 21% O₂ (normoxia). Cellular functions were assessed by flow cytometry, Transwell and Cell Counting Kit‑8 assays in LDHA‑silenced ESCs and Ishikawa cells. Mitochondrial functions were evaluated using mitochondrial membrane potential JC‑1 staining, reactive oxygen species flow cytometric analysis and ATP detection. Additionally, lactic acid production was examined and western blotting was used to evaluate the expression levels of proteins associated with apoptosis, cell cycle and glycolysis, as well as regulatory proteins involved in epithelial‑mesenchymal transformation and glycolytic pathways. LDHA was localized to endometrial glandular cells and stromal cells. However, LDHA protein expression was higher in endometriotic lesions compared with that in normal and eutopic endometria. LDHA expression levels in ectopic glandular cells were higher during the proliferative stage compared with during the secretory stage. Hypoxia treatment of Ishikawa cells and ESCs markedly induced the mRNA and protein expression of LDHA. Silencing of LDHA expression in Ishikawa cells and THESC cells significantly promoted impaired mitochondrial function and apoptosis while inhibiting migration and glycolysis. However, it had no obvious effect on proliferation. In conclusion, the present study revealed that LDHA was highly expressed in endometriotic tissues, where it may serve a notable role in the occurrence and development of endometriosis.

View Figures

View References

1	Bulun SE: Endometriosis. N Engl J Med. 360:268–279. 2009. View Article : Google Scholar : PubMed/NCBI
2	Batt RE; Invited comment on the paper by Benagiano, ; et al: Entitled ‘the history of endometriosis’. Gynecol Obstet Invest. 78:10–11. 2014. View Article : Google Scholar : PubMed/NCBI
3	Shubina AN, Egorova AA, Baranov VS and Kiselev AV: Recent advances in gene therapy of endometriosis. Recent Pat DNA Gene Seq. 7:169–178. 2013. View Article : Google Scholar : PubMed/NCBI
4	Giudice LC and Kao LC: Endometriosis. Lancet. 364:1789–1799. 2004. View Article : Google Scholar : PubMed/NCBI
5	Wu MH, Chen KF, Lin SC, Lgu CW and Tsai SJ: Aberrant expression of leptin in human endometriotic stromal cells is induced by elevated levels of hypoxia inducible factor-1alpha. Am J Pathol. 170:590–598. 2007. View Article : Google Scholar : PubMed/NCBI
6	Hsiao KY, Lin SC, Wu MH and Tsai SJ: Pathological functions of hypoxia in endometriosis. Front Biosci (Elite Ed). 7:309–321. 2015.PubMed/NCBI
7	Lin X, Dai Y, Xu W, Shi L, Jin X, Li C, Zhou F, Pan Y, Zhang Y, Lin X and Zhang S: Hypoxia promotes ectopic adhesion ability of endometrial stromal cells via TGF-β1/smad signaling in endometriosis. Endocrinology. 159:1630–1641. 2018. View Article : Google Scholar : PubMed/NCBI
8	Dai Y, Lin X, Xu W, Lin X, Huang Q, Shi L, Pan Y, Zhang Y, Zhu Y, Li C, et al: MiR-210-3p protects endometriotic cells from oxidative stress-induced cell cycle arrest by targeting BARD1. Cell Death Dis. 10:1442019. View Article : Google Scholar : PubMed/NCBI
9	Young VJ, Brown JK, Maybin J, Saunders PT, Duncan WC and Horne AW: Transforming growth factor-β induced Warburg-like metabolic reprogramming may underpin the development of peritoneal endometriosis. J Clin Endocrinol Metab. 99:3450–3459. 2014. View Article : Google Scholar : PubMed/NCBI
10	Molinari E, Bar H, Pyle AM and Patrizio P: Transcriptome analysis of human cumulus cells reveals hypoxia as the main determinant of follicular senescence. Mol Hum Reprod. 22:866–876. 2016. View Article : Google Scholar : PubMed/NCBI
11	Lee HC, Lin SC, Wu MH and Tsai SJ: Induction of pyruvate dehydrogenase kinase 1 by hypoxia alters cellular metabolism and inhibits apoptosis in endometriotic stromal cells. Reprod Sci. 26:734–744. 2019. View Article : Google Scholar : PubMed/NCBI
12	Rong Y, Wu W, Ni X, Kuang T, Jin D, Wang D and Lou W: Lactate dehydrogenase A is overexpressed in pancreatic cancer and promotes the growth of pancreatic cancer cells. Tumour Biol. 34:1523–1530. 2013. View Article : Google Scholar : PubMed/NCBI
13	Koukourakis MI, Giatromanolaki A, Winter S, Leek R, Sivridis E and Harris AL: Lactate dehydrogenase 5 expression in squamous cell head and neck cancer relates to prognosis following radical or postoperative radiotherapy. Oncology. 77:285–292. 2009. View Article : Google Scholar : PubMed/NCBI
14	Giatromanolaki A, Koukourakis MI, Pezzella F, Sivridis E, Turley H, Harris AL and Gatter KC: Lactate dehydrogenase 5 expression in non-Hodgkin B-cell lymphomas is associated with hypoxia regulated proteins. Leuk Lymphoma. 49:2181–2186. 2008. View Article : Google Scholar : PubMed/NCBI
15	Miao P, Sheng S, Sun X, Liu J and Huang G: Lactate dehydrogenase A in cancer: A promising target for diagnosis and therapy. IUBMB Life. 65:904–910. 2013. View Article : Google Scholar : PubMed/NCBI
16	Semenza GL: HIF-1 mediates metabolic responses to intratumoral hypoxia and oncogenic mutations. J Clin Invest. 123:3664–3671. 2013. View Article : Google Scholar : PubMed/NCBI
17	Yeung SJ, Pan J and Lee MH: Roles of p53, MYC and HIF-1 in regulating glycolysis-the seventh hallmark of cancer. Cell Mol Life Sci. 65:3981–3999. 2008. View Article : Google Scholar : PubMed/NCBI
18	Holmes RS and Goldberg E: Computational analyses of mammalian lactate dehydrogenases: Human, mouse, opossum and platypus LDHs. Comput Biol Chem. 33:379–385. 2009. View Article : Google Scholar : PubMed/NCBI
19	Zhan L, Wang W, Zhang Y, Song E, Fan Y and Wei B: Hypoxia-inducible factor-1alpha: A promising therapeutic target in endometriosis. Biochimie. 123:130–137. 2016. View Article : Google Scholar : PubMed/NCBI
20	Griffiths EA, Pritchard SA, Valentine HR, Whitchelo N, Bishop PW, Ebert MP, Price PM, Welch IM and West CM: Hypoxia-inducible factor-1alpha expression in the gastric carcinogenesis sequence and its prognostic role in gastric and gastro-oesophageal adenocarcinomas. Br J Cancer. 96:95–103. 2007. View Article : Google Scholar : PubMed/NCBI
21	Sun HC, Qiu ZJ, Liu J, Sun J, Jiang T, Huang KJ, Yao M and Huang C: Expression of hypoxia-inducible factor-1 alpha and associated proteins in pancreatic ductal adenocarcinoma and their impact on prognosis. Int J Oncol. 30:1359–1367. 2007.PubMed/NCBI
22	Maybin JA, Hirani N, Brown P, Jabbour HN and Critchley HO: The regulation of vascular endothelial growth factor by hypoxia and prostaglandin F2α during human endometrial repair. J Clin Endocrinol Metab. 96:2475–2483. 2011. View Article : Google Scholar : PubMed/NCBI
23	Maybin JA, Hirani N, Jabbour HN and Critchley HO: Novel roles for hypoxia and prostaglandin E2 in the regulation of IL-8 during endometrial repair. Am J Pathol. 178:1245–1256. 2011. View Article : Google Scholar : PubMed/NCBI
24	D'Amico F, Skarmoutsou E, Quaderno G, Malaponte G, La Corte C, Scibilia G, D'Agate G, Scollo P, Fraggetta F, Spandidos DA and Mazzarino MC: Expression and localisation of osteopontin and prominin-1 (CD133) in patients with endometriosis. Int J Mol Med. 31:1011–1016. 2013. View Article : Google Scholar
25	He G, Jiang Y, Zhang B and Wu G: The effect of HIF-1alpha on glucose metabolism, growth and apoptosis of pancreatic cancerous cells. Asia Pac J Clin Nutr. 23:174–180. 2014.PubMed/NCBI
26	Revised American Society for Reproductive Medicine classification of endometriosis, . 1996. Fertil Steril. 67:817–821. 1997. View Article : Google Scholar : PubMed/NCBI
27	Ryan IP, Schriock ED and Taylor RN: Isolation, characterization, and comparison of human endometrial and endometriosis cells in vitro. J Clin Endocrinol Metab. 78:642–649. 1994. View Article : Google Scholar : PubMed/NCBI
28	Hsiao KY, Wu MH, Chang N, Yang SH, Wu CW, Sun HS and Tsai SJ: Coordination of AUF1 and miR-148a destabilizes DNA methyltransferase 1 mRNA under hypoxia in endometriosis. Mol Hum Reprod. 21:894–904. 2015. View Article : Google Scholar : PubMed/NCBI
29	Hsiao KY, Chang N, Lin SC, Li YH and Wu MH: Inhibition of dual specificity phosphatase-2 by hypoxia promotes interleukin-8-mediated angiogenesis in endometriosis. Hum Reprod. 29:2747–2755. 2014. View Article : Google Scholar : PubMed/NCBI
30	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
31	Chiappini F, Baston JI, Vaccarezza A, Singla JJ, Pontillo C, Miret N, Farina M, Meresman G and Randi A: Enhanced cyclooxygenase-2 expression levels and metalloproteinase 2 and 9 activation by Hexachlorobenzene in human endometrial stromal cells. Biochem Pharmacol. 109:91–104. 2016. View Article : Google Scholar : PubMed/NCBI
32	Wang D, Luo Y, Wang G and Yang Q: CircATRNL1 promotes epithelial-mesenchymal transition in endometriosis by upregulating Yes-associated protein 1 in vitro. Cell Death Dis. 11:5942020. View Article : Google Scholar : PubMed/NCBI
33	Cui XG, Han ZT, He SH, Wu XD, Chen TR, Shao CH, Chen DL, Su N, Chen YM, Wang T, et al: HIF1/2α mediates hypoxia-induced LDHA expression in human pancreatic cancer cells. Oncotarget. 8:24840–24852. 2017. View Article : Google Scholar : PubMed/NCBI
34	Abate M, Festa A, Falco M, Lombardi A, Luce A, Grimaldi A, Zappavigna S, Sperlongano P, Irace C, Caraglia M, et al: Mitochondria as playmakers of apoptosis, autophagy and senescence. Semin Cell Dev Biol. 98:139–153. 2020. View Article : Google Scholar : PubMed/NCBI
35	Chen J, Liao W, Gao W, Huang J and Gao Y: Intermittent hypoxia protects cerebral mitochondrial function from calcium overload. Acta Neurol Belg. 113:507–513. 2013. View Article : Google Scholar : PubMed/NCBI
36	Wu MH, Hsiao KY and Tsai SJ: Hypoxia: The force of endometriosis. J Obstet Gynaecol Res. 45:532–541. 2019. View Article : Google Scholar : PubMed/NCBI
37	Kasvandik S, Samuel K, Peters M, Eimre M, Peet N, Roost AM, Padrik L, Paju K, Peil L and Salumets A: Deep quantitative proteomics reveals extensive metabolic reprogramming and cancer-like changes of ectopic endometriotic stromal cells. J Proteome Res. 15:572–584. 2016. View Article : Google Scholar : PubMed/NCBI
38	Gebel HM, Braun DP, Tambur A, Frame D, Rana N and Dmowski WP: Spontaneous apoptosis of endometrial tissue is impaired in women with endometriosis. Fertil Steril. 69:1042–1047. 1998. View Article : Google Scholar : PubMed/NCBI
39	Vaskivuo TE, Stenback F, Karhumaa P, Risteli J, Dunkel L and Tapanainen JS: Apoptosis and apoptosis-related proteins in human endometrium. Mol Cell Endocrinol. 165:75–83. 2000. View Article : Google Scholar : PubMed/NCBI
40	Singh R, Letai A and Sarosiek K: Regulation of apoptosis in health and disease: The balancing act of BCL-2 family proteins. Nat Rev Mol Cell Biol. 20:175–193. 2019. View Article : Google Scholar : PubMed/NCBI
41	Oltvai ZN and Korsmeyer SJ: Checkpoints of dueling dimers foil death wishes. Cell. 79:189–192. 1994. View Article : Google Scholar : PubMed/NCBI
42	Harada M, Suganuma N, Furuhashi M, Nagasaka T, Nakashima N, Kikkawa F, Tomoda Y and Furui K: Detection of apoptosis in human endometriotic tissues. Mol Hum Reprod. 2:307–315. 1996. View Article : Google Scholar : PubMed/NCBI
43	Meresman GF, Vighi S, Buquet RA, Contreras-Ortiz O, Tesone M and Rumi LS: Apoptosis and expression of Bcl-2 and Bax in eutopic endometrium from women with endometriosis. Fertil Steril. 74:760–766. 2000. View Article : Google Scholar : PubMed/NCBI
44	McLaren J, Prentice A, Charnock-Jones DS, Sharkey AM and Smith SK: Immunolocalization of the apoptosis regulating proteins Bcl-2 and Bax in human endometrium and isolated peritoneal fluid macrophages in endometriosis. Hum Reprod. 12:146–152. 1997. View Article : Google Scholar : PubMed/NCBI
45	Depalo R, Cavallini A, Lorusso F, Bassi E, Totaro I, Marzullo A, Bettocchi S and Selvaggi L: Apoptosis in normal ovaries of women with and without endometriosis. Reprod Biomed Online. 19:808–815. 2009. View Article : Google Scholar : PubMed/NCBI
46	Zorov DB, Juhaszova M and Sollott SJ: Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev. 94:909–950. 2014. View Article : Google Scholar : PubMed/NCBI
47	Bulthuis EP, Adjobo-Hermans MJW, Willems P and Koopman WJH: Mitochondrial morphofunction in mammalian cells. Antioxid Redox Signal. 30:2066–2109. 2019. View Article : Google Scholar : PubMed/NCBI
48	Liu L, Feng D, Chen G, Chen M, Zheng Q, Song P, Ma Q, Zhu C, Wang R, Qi W, et al: Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells. Nat Cell Biol. 14:177–185. 2012. View Article : Google Scholar : PubMed/NCBI
49	Nagafuchi A, Shirayoshi Y, Okazaki K, Yasuda K and Takeichi M: Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 329:341–343. 1987. View Article : Google Scholar : PubMed/NCBI
50	Saliminejad K, Edalatkhah H, Kamali K, Memariani T, Nasiri M, Saket M and Khorram Khorshid HR: Association of common variations of the E-cadherin with endometriosis. Gynecol Endocrinol. 31:899–902. 2015. View Article : Google Scholar : PubMed/NCBI
51	Ma T, Fu B, Yang X, Xiao Y and Pan M: Adipose mesenchymal stem cell-derived exosomes promote cell proliferation, migration, and inhibit cell apoptosis via Wnt/beta-catenin signaling in cutaneous wound healing. J Cell Biochem. 120:10847–10854. 2019. View Article : Google Scholar : PubMed/NCBI
52	Hinck L, Nathke IS, Papkoff J and Nelson WJ: Dynamics of cadherin/catenin complex formation: Novel protein interactions and pathways of complex assembly. J Cell Biol. 125:1327–1340. 1994. View Article : Google Scholar : PubMed/NCBI
53	Zhang X, Liu Z, Chen M, Cao Q and Huang D: Effects of S100A6 gene silencing on the biological features of eutopic endometrial stromal cells and betacatenin expression. Mol Med Rep. 15:1279–1285. 2017. View Article : Google Scholar : PubMed/NCBI
54	Zhao J, Huang X, Xu Z, Dai J, He H, Zhu Y and Wang H: LDHA promotes tumor metastasis by facilitating epithelialmesenchymal transition in renal cell carcinoma. Mol Med Rep. 16:8335–8344. 2017. View Article : Google Scholar : PubMed/NCBI