Effect of Long Non-Coding RNA-02075 on Glycolysis in Endometrium Stromal Cells of Endometriosis: A Preliminary Experimental Study

doi:10.12280/gjszjk.20230047

Abstract

Abstract:

Objective: To compare the expression levels of long non-coding RNA-02075 (LINC02075) in the eutopic and ectopic endometrium from patients with endometriosis, and to investigate the possible roles of LINC02075 in the regulation of glycolysis in endometrial stromal cells. Methods: The samples of eutopic and ectopic endometrium were from 10 patients with endometriosis aged 23-36 years. The endometrium samples from 10 normal women accepting the surgery of intrauterine device placement aged 25-36 years were used as the control. The expression of LINC02075 was evaluated by RT-qPCR. The ectopic endometrial stromal cells were in vitro cultured. The overexpression of LINC02075 was induced by the lentivirus and adenovirus vectors. The glycolytic enzymes, cell proliferation, migration and apoptosis rates were tested. Results: LINC02075 was widely expressed in the nucleus and cytoplasm of endometrial stromal cells. The expression level of LINC02075 mRNA in ectopic endometrium (the endometriosis group) was significantly higher than that in normal endometrium (the control group) and eutopic endometrium (the endometriosis group) (all P<0.05). Overexpression of LINC02075 significantly increased glucose consumption (P<0.05), and increased lactate production (but P>0.05) in ectopic endothelial stromal cells, while the expression levels of glycolysis-related enzymes such as hexokinase 1 (HK1), pyruvate dehydrogenase kinase 1 (PDK1), lactate dehydrogenase A (LDHA), glucose transporter type 1 (GLUT1) and aldolase A (ALDOA) were also increased, among which the increased expression level of LDHA mRNA was statistically significant (P<0.05). Overexpressed LINC02075 also increased the proliferation and apoptosis and decreased the migration of endometrial stromal cells (all P<0.05). Conclusions: The abnormally elevated LINC02075 in ectopic endometrium of patients with endometriosis may promote proliferation and reduce migration of endometrial stromal cells by the enhanced glycolytic mechanism.

Key words: Endometriosis, Cell proliferation, Warburg effect, oncologic, Aerobic glycolysis, LINC02075

ZHU Pei-pei, LI Kai-quan, ZHONG Chen-yi, SHU Li, WU Chun-xiang, LIU Jin-yong, HOU Zhen, DIAO Fei-yang, MAO Yun-dong. Effect of Long Non-Coding RNA-02075 on Glycolysis in Endometrium Stromal Cells of Endometriosis: A Preliminary Experimental Study[J]. Journal of International Reproductive Health/Family Planning, 2023, 42(4): 265-271.

Figures/Tables 5

References 27

[1]	Wei Y, Liang Y, Lin H, et al. Autonomic nervous system and inflammation interaction in endometriosis-associated pain[J]. J Neuroinflammation, 2020, 17(1):80. doi: 10.1186/s12974-020-01752-1. doi: 10.1186/s12974-020-01752-1
[2]	Chapron C, Marcellin L, Borghese B, et al. Rethinking mechanisms, diagnosis and management of endometriosis[J]. Nat Rev Endocrinol, 2019, 15(11):666-682. doi: 10.1038/s41574-019-0245-z. doi: 10.1038/s41574-019-0245-z pmid: 31488888
[3]	Sampson JEA. Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity[J]. Am J Obstet Gynecol, 1927, 14(4):93-94. doi: 10.1016/S0002-9378(15)30003-X. doi: 10.1016/S0002-9378(15)30003-X
[4]	Moen MH, Magnus P. The familial risk of endometriosis[J]. Acta Obstet Gynecol Scand, 1993, 72(7):560-564. doi: 10.3109/00016349309058164. doi: 10.3109/00016349309058164
[5]	Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation[J]. Cell, 2011, 144(5):646-674. doi: 10.1016/j.cell.2011.02.013. doi: 10.1016/j.cell.2011.02.013 pmid: 21376230
[6]	Richart L, Picod-Chedotel ML, Wassef M, et al. XIST loss impairs mammary stem cell differentiation and increases tumorigenicity through Mediator hyperactivation[J]. Cell, 2022, 185(12):2164-2183.e25. doi: 10.1016/j.cell.2022.04.034. doi: 10.1016/j.cell.2022.04.034 pmid: 35597241
[7]	Wang R, Ma Z, Feng L, et al. LncRNA MIR31HG targets HIF1A and P21 to facilitate head and neck cancer cell proliferation and tumorigenesis by promoting cell-cycle progression[J]. Mol Cancer, 2018, 17(1):162. doi: 10.1186/s12943-018-0916-8. doi: 10.1186/s12943-018-0916-8 pmid: 30458787
[8]	Pandey RR, Mondal T, Mohammad F, et al. Kcnq1ot1 antisense noncoding RNA mediates lineage-specific transcriptional silencing through chromatin-level regulation[J]. Mol Cell, 2008, 32(2):232-246. doi: 10.1016/j.molcel.2008.08.022. doi: 10.1016/j.molcel.2008.08.022 pmid: 18951091
[9]	Tsai MC, Manor O, Wan Y, et al. Long noncoding RNA as modular scaffold of histone modification complexes[J]. Science, 2010, 329(5992):689-693. doi: 10.1126/science.1192002. doi: 10.1126/science.1192002 URL
[10]	Gong C, Maquat LE. lncRNAs transactivate STAU1-mediated mRNA decay by duplexing with 3′ UTRs via Alu elements[J]. Nature, 2011, 470(7333):284-288. doi: 10.1038/nature09701. doi: 10.1038/nature09701
[11]	Tripathi V, Ellis JD, Shen Z, et al. The nuclear-retained noncoding RNA MALAT1 regulates alternative splicing by modulating SR splicing factor phosphorylation[J]. Mol Cell, 2010, 39(6):925-938. doi: 10.1016/j.molcel.2010.08.011. doi: 10.1016/j.molcel.2010.08.011 pmid: 20797886
[12]	Wang C, Li Y, Yan S, et al. Interactome analysis reveals that lncRNA HULC promotes aerobic glycolysis through LDHA and PKM2[J]. Nat Commun, 2020, 11(1):3162. doi: 10.1038/s41467-020-16966-3. doi: 10.1038/s41467-020-16966-3 pmid: 32572027
[13]	Wang X, Zhang J, Liu X, et al. Long noncoding RNAs in endometriosis: Biological functions, expressions, and mechanisms[J]. J Cell Physiol, 2021, 236(1):6-14. doi: 10.1002/jcp.29847. doi: 10.1002/jcp.29847 URL
[14]	Qi X, Zhang Y, Ji H, et al. Knockdown of prohibitin expression promotes glucose metabolism in eutopic endometrial stromal cells from women with endometriosis[J]. Reprod Biomed Online, 2014, 29(6):761-770. doi: 10.1016/j.rbmo.2014.09.004. doi: 10.1016/j.rbmo.2014.09.004 pmid: 25444511
[15]	Fang Y, Fullwood MJ. Roles, Functions, and Mechanisms of Long Non-coding RNAs in Cancer[J]. Genomics Proteomics Bioinformatics, 2016, 14(1):42-54. doi: 10.1016/j.gpb.2015.09.006. doi: 10.1016/j.gpb.2015.09.006 URL
[16]	Jia H, Osak M, Bogu GK, et al. Genome-wide computational identification and manual annotation of human long noncoding RNA genes[J]. RNA, 2010, 16(8):1478-1487. doi: 10.1261/rna.1951310. doi: 10.1261/rna.1951310 pmid: 20587619
[17]	Lu C, Qiao P, Fu R, et al. Phosphorylation of PFKFB4 by PIM2 promotes anaerobic glycolysis and cell proliferation in endometriosis[J]. Cell Death Dis, 2022, 13(9):790. doi: 10.1038/s41419-022-05241-6. doi: 10.1038/s41419-022-05241-6 pmid: 36109523
[18]	Zhu Y, Jin L, Shi R, et al. The long noncoding RNA glycoLINC assembles a lower glycolytic metabolon to promote glycolysis[J]. Mol Cell, 2022, 82(3):542-554.e6. doi: 10.1016/j.molcel.2021.11.017. doi: 10.1016/j.molcel.2021.11.017 pmid: 35081364
[19]	Park MK, Zhang L, Min KW, et al. NEAT1 is essential for metabolic changes that promote breast cancer growth and metastasis[J]. Cell Metab, 2021, 33(12):2380-2397.e9. doi: 10.1016/j.cmet.2021.11.011. doi: 10.1016/j.cmet.2021.11.011 pmid: 34879239
[20]	Cui D, Ma J, Liu Y, et al. Analysis of long non-coding RNA expression profiles using RNA sequencing in ovarian endometriosis[J]. Gene, 2018, 673:140-148. doi: 10.1016/j.gene.2018.06.046. doi: S0378-1119(18)30702-9 pmid: 29920364
[21]	Xue L, Li J, Lin Y, et al. m⁶A transferase METTL3-induced lncRNA ABHD11-AS1 promotes the Warburg effect of non-small-cell lung cancer[J]. J Cell Physiol, 2021, 236(4):2649-2658. doi: 10.1002/jcp.30023. doi: 10.1002/jcp.30023 URL
[22]	Warburg O. The Metabolism of Carcinoma Cells1[J]. J Cancer Res, 1925, 9(1):148-163. doi: 10.1158/jcr.1925.148. doi: 10.1158/jcr.1925.148
[23]	Li B, Kang H, Xiao Y, et al. LncRNA GAL promotes colorectal cancer liver metastasis through stabilizing GLUT1[J]. Oncogene, 2022, 41(13):1882-1894. doi: 10.1038/s41388-022-02230-z. doi: 10.1038/s41388-022-02230-z pmid: 35149838
[24]	Zhao S, Guan B, Mi Y, et al. LncRNA MIR17HG promotes colorectal cancer liver metastasis by mediating a glycolysis-associated positive feedback circuit[J]. Oncogene, 2021, 40(28):4709-4724. doi: 10.1038/s41388-021-01859-6. doi: 10.1038/s41388-021-01859-6 pmid: 34145399
[25]	Diao R, Wei W, Zhao J, et al. CCL19/CCR7 contributes to the pathogenesis of endometriosis via PI3K/Akt pathway by regulating the proliferation and invasion of ESCs[J]. Am J Reprod Immunol,2017 Nov; 78(5). doi: 10.1111/aji.12744. doi: 10.1111/aji.12744
[26]	Hou XX, Zhou WJ, Wang XQ, et al. Fractalkine/CX3CR1 is involved in the pathogenesis of endometriosis by regulating endometrial stromal cell proliferation and invasion[J]. Am J Reprod Immunol, 2016, 76(4):318-325. doi: 10.1111/aji.12557. doi: 10.1111/aji.12557 URL
[27]	Lee HC, Lin SC, Wu MH, et al. Induction of Pyruvate Dehydrogenase Kinase 1 by Hypoxia Alters Cellular Metabolism and Inhibits Apoptosis in Endometriotic Stromal Cells[J]. Reprod Sci, 2019, 26(6):734-744. doi: 10.1177/1933719118789513. doi: 10.1177/1933719118789513 URL

引物名称	引物序列（5′-3′）
LINC02075	上游：CTGAGACCATCAACCCTATC
	下游：ACTTATTCAACTGGCACTCC
GLUT1	上游：GGCCAAGAGTGTGCTAAAGAA
	下游：ACAGCGTTGATGCCAGACAG
PDK1	上游：TGTGAAGATGAGTGACCGAGGAG
	下游：GGCAATCCATAACCAAAACCAG
HK1	上游：GCTCTCCGATGAAACTCTCATAG
	下游：GGACCTTACGAATGTTGGCAA
PKM	上游：ATGTCGAAGCCCCATAGTGAA
	下游：TGGGTGGTGAATCAATGTCCA
ALDOA	上游：ATGTCGAAGCCCCATAGTGAA
	下游：TGGGTGGTGAATCAATGTCCA
LDHA	上游：ATGGCAACTCTAAAGGATCAGC
	下游：CCAACCCCAACAACTGTAATCT
SDHA	上游：CGGCATTCCCACCAACTA
	下游：GCCCGACCAAAGACAACC
β-actin	上游：TCAAGATCATTGCTCCTCCTGAG
	下游：ACATCTGCTGGAAGGTGGACA

引物名称	引物序列（5′-3′）
LINC02075	上游：CTGAGACCATCAACCCTATC
	下游：ACTTATTCAACTGGCACTCC
GLUT1	上游：GGCCAAGAGTGTGCTAAAGAA
	下游：ACAGCGTTGATGCCAGACAG
PDK1	上游：TGTGAAGATGAGTGACCGAGGAG
	下游：GGCAATCCATAACCAAAACCAG
HK1	上游：GCTCTCCGATGAAACTCTCATAG
	下游：GGACCTTACGAATGTTGGCAA
PKM	上游：ATGTCGAAGCCCCATAGTGAA
	下游：TGGGTGGTGAATCAATGTCCA
ALDOA	上游：ATGTCGAAGCCCCATAGTGAA
	下游：TGGGTGGTGAATCAATGTCCA
LDHA	上游：ATGGCAACTCTAAAGGATCAGC
	下游：CCAACCCCAACAACTGTAATCT
SDHA	上游：CGGCATTCCCACCAACTA
	下游：GCCCGACCAAAGACAACC
β-actin	上游：TCAAGATCATTGCTCCTCCTGAG
	下游：ACATCTGCTGGAAGGTGGACA