Restoration of microRNA‑29c in type I endometrioid cancer reduced endometrial cancer cell growth

Van Sinderen,Michelle; Griffiths,Meaghan; Menkhorst,Ellen; Niven,Keith; Dimitriadis,Evdokia

doi:10.3892/ol.2019.10588

Restoration of microRNA‑29c in type I endometrioid cancer reduced endometrial cancer cell growth

Authors:
- Michelle Van Sinderen
- Meaghan Griffiths
- Ellen Menkhorst
- Keith Niven
- Evdokia Dimitriadis
View Affiliations

Affiliations: Embryo Implantation Laboratory, Centre for Reproductive Health, Hudson Institute of Medical Research, Clayton, Victoria 3186, Australia, FlowCore, Technology Research Platforms, Monash University, Clayton, Victoria 3800, Australia
Published online on: July 9, 2019 https://doi.org/10.3892/ol.2019.10588
Pages: 2684-2693

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

Endometrial cancer is the most common gynaecological cancer worldwide, and the prognosis of patients with advanced disease remains poor. MicroRNAs (miRs) are dysregulated in endometrial cancer. miRs‑29‑a, ‑b and ‑c expression levels are downregulated in endometrial cancer; however, a specific role for miR‑29c and its target genes remain to be elucidated. The aim of the present study was to determine the functional effect of restoring miR‑29c expression in endometrial cancer cell lines and to identify miR‑29c targets involved in cancer progression. miR‑29c expression in human endometrial tumour grades 1‑3 and benign tissue as well as in the endometrial cancer cell lines Ishikawa, HEC1A and AN3CA were analysed using reverse transcriptase‑quantitative PCR (RT‑qPCR). The cell lines were transfected with miR‑29c mimic, miR‑29c inhibitor or scrambled control. xCELLigence real‑time cell monitoring analysed proliferation and migration, and flow cytometry was used to analyse apoptosis and cell cycle. The expression of miR‑29c target genes in transfected cell lines was analysed using RT‑qPCR. miR‑29c was downregulated in grade 1‑3 endometrial cancer samples compared with benign endometrium. miR‑29c was reduced in Ishikawa and AN3CA cells, but not in HEC1A cell lines compared with non‑cancerous primary human endometrial epithelial cells. Overexpression of miR‑29c variably reduced proliferation, increased apoptosis and reduced the expression levels of miR‑29c target genes, including cell division cycle 42, HMG‑box transcription factor 1, integrin subunit β 1, MCL1 apoptosis regulator BCL2 family member, MDM2 proto‑oncogene, serum/glucocorticoid regulated kinase 1, sirtuin 1 and vascular endothelial growth factor A, across the three cell lines investigated. Inhibition of miR‑29c in HEC1A cells increased proliferation and collagen type IV α 1 chain expression. The re‑introduction of miR‑29c to endometrial cancer cell lines reduced proliferation, increased apoptosis and reduced miR‑29c target gene expression in vitro. The present results suggested that miR‑29c may be a potential therapeutic target for endometrial cancer.

View Figures

View References

1	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar : PubMed/NCBI
2	Soliman PT, Oh JC, Schmeler KM, Sun CC, Slomovitz BM, Gershenson DM, Burke TW and Lu KH: Risk factors for young premenopausal women with endometrial cancer. Obstet Gynecol. 105:575–580. 2005. View Article : Google Scholar : PubMed/NCBI
3	Bilyk O, Coatham M, Jewer M and Postovit LM: Epithelial-to-Mesenchymal Transition in the female reproductive tract: From normal function to disease pathology. Front Oncol. 7:1452017. View Article : Google Scholar : PubMed/NCBI
4	Hong B, Le Gallo M and Bell DW: The mutational landscape of endometrial cancer. Curr Opin Genet Dev. 30:25–31. 2015. View Article : Google Scholar : PubMed/NCBI
5	Di Cristofano A and Ellenson LH: Endometrial carcinoma. Annu Rev Pathol. 2:57–85. 2007. View Article : Google Scholar : PubMed/NCBI
6	Piulats JM, Guerra E, Gil-Martin M, Roman-Canal B, Gatius S, Sanz-Pamplona R, Velasco A, Vidal A and Matias-Guiu X: Molecular approaches for classifying endometrial carcinoma. Gynecol Oncol. 145:200–207. 2017. View Article : Google Scholar : PubMed/NCBI
7	Arend RC, Jones BA, Martinez A and Goodfellow P: Endometrial cancer: Molecular markers and management of advanced stage disease. Gynecol Oncol. 150:569–580. 2018. View Article : Google Scholar : PubMed/NCBI
8	Kim VN: MicroRNA biogenesis: Coordinated cropping and dicing. Nat Rev Mol Cell Biol. 6:376–385. 2005. View Article : Google Scholar : PubMed/NCBI
9	Schmitt MJ, Margue C, Behrmann I and Kreis S: MiRNA-29: A microRNA family with tumor-suppressing and immune-modulating properties. Curr Mol Med. 13:572–585. 2013. View Article : Google Scholar : PubMed/NCBI
10	Castilla MÁ, Moreno-Bueno G, Romero-Pérez L, Van De Vijver K, Biscuola M, López-García MÁ, Prat J, Matías-Guiu X, Cano A, Oliva E and Palacios J: Micro-RNA signature of the epithelial-mesenchymal transition in endometrial carcinosarcoma. J Pathol. 223:72–80. 2011. View Article : Google Scholar : PubMed/NCBI
11	Chen HX, Xu XX, Tan BZ, Zhang Z and Zhou XD: Micro-RNA-29b inhibits angiogenesis by targeting VEGFA through the MAPK/ERK and PI3K/Akt signaling pathways in endometrial carcinoma. Cell Physiol Biochem. 41:933–946. 2017. View Article : Google Scholar : PubMed/NCBI
12	Hiroki E, Akahira J, Suzuki F, Nagase S, Ito K, Suzuki T, Sasano H and Yaegashi N: Changes in microRNA expression levels correlate with clinicopathological features and prognoses in endometrial serous adenocarcinomas. Cancer Sci. 101:241–249. 2010. View Article : Google Scholar : PubMed/NCBI
13	Jiang T, Sui D, You D, Yao S, Zhang L, Wang Y, Zhao J and Zhang Y: miR-29a-5p inhibits proliferation and invasion and induces apoptosis in endometrial carcinoma via targeting TPX2. Cell Cycle. 17:1268–1278. 2018. View Article : Google Scholar : PubMed/NCBI
14	Ushakov DS, Dorozhkova AS, Babayants EV, Ovchinnikov VY, Kushlinskii DN, Adamyan LV, Gulyaeva LF and Kushlinskii NE: Expression of microRNA potentially regulated by AhR and CAR in malignant tumors of the endometrium. Bull Exp Biol Med. 165:688–691. 2018. View Article : Google Scholar : PubMed/NCBI
15	Mazzoccoli L, Robaina MC, Apa AG, Bonamino M, Pinto LW, Queiroga E, Bacchi CE and Klumb CE: MiR-29 silencing modulates the expression of target genes related to proliferation, apoptosis and methylation in Burkitt lymphoma cells. J Cancer Res Clin Oncol. 144:483–497. 2018. View Article : Google Scholar : PubMed/NCBI
16	Griffiths M, Van Sinderen M, Rainczuk K and Dimitriadis E: miR-29c overexpression and COL4A1 downregulation in infertile human endometrium reduces endometrial epithelial cell adhesive capacity in vitro implying roles in receptivity. Sci Rep. 9:86442019. View Article : Google Scholar : PubMed/NCBI
17	Long M, Wan X, La X, Gong X and Cai X: miR-29c is downregulated in the ectopic endometrium and exerts its effects on endometrial cell proliferation, apoptosis and invasion by targeting c-Jun. Int J Mol Med. 35:1119–1125. 2015. View Article : Google Scholar : PubMed/NCBI
18	Marwood M, Visser K, Salamonsen LA and Dimitriadis E: Interleukin-11 and leukemia inhibitory factor regulate the adhesion of endometrial epithelial cells: Implications in fertility regulation. Endocrinology. 150:2915–2923. 2009. View Article : Google Scholar : PubMed/NCBI
19	Winship A, Van Sinderen M, Rainczuk K and Dimitriadis E: Therapeutically blocking Interleukin-11 Receptor-alpha enhances doxorubicin cytotoxicity in high grade type I endometrioid tumours. Oncotarget. 8:22716–22729. 2017. View Article : Google Scholar : PubMed/NCBI
20	Van Sinderen M, Cuman C, Winship A, Menkhorst E and Dimitriadis E: The chrondroitin sulfate proteoglycan (CSPG4) regulates human trophoblast function. Placenta. 34:907–912. 2013. View Article : Google Scholar : PubMed/NCBI
21	Livak K and Schmittgen T: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta DeltaC(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
22	Licholai S, Szczekilk W and Sanak M: miR-29c-3p is an effective biomarker of abdominal aortic aneurysm in patients undergoing elective surgery. MicroRNA. 5:124–131. 2016. View Article : Google Scholar : PubMed/NCBI
23	Xu Z, Zou L, Gang M, Wu X, Huang F, Feng T, Li S, Lin Q, He X, Liu Z and Cao X: Integrin b1 is a critical effector in promoting metastasis and chemo-resistance of esophageal squamous cell carcinoma. Am J Cancer Res. 7:531–542. 2017.PubMed/NCBI
24	Gupta A, Shah K, Oza MJ and Behl T: Reactivation of p53 gene by MDM2 inhibitors: a novel therapy for cancer treatment. Biomed Pharmacother. 109:484–492. 2019. View Article : Google Scholar : PubMed/NCBI
25	Weigelt B, Warne PH, Lambros MB, Reis-Filho JS and Downward J: PI3K pathway dependencies in endometrial cancer cell lines. Clin Cancer Res. 19:3533–3544. 2013. View Article : Google Scholar : PubMed/NCBI
26	Bhardwaj A, Singh H, Rajapakshe K, Tachibana K, Ganesan N, Pan Y, Gunaratne PH, Coarfa C and Bedrosian I: Regulation of miRNA-29c and its downstream pathways in preneoplastic progression of triple-negative breast cancer. Oncotarget. 8:19645–19660. 2017. View Article : Google Scholar : PubMed/NCBI
27	Chen G, Zhou T, Li Y, Yu Z and Sun L: p53 target miR-29c-3p suppresses colon cancer cell invasion and migration through inhibition of PHLDB2. Biochem Biophys Res Commun. 487:90–95. 2017. View Article : Google Scholar : PubMed/NCBI
28	Chuang TD and Khorram O: Mechanisms underlying aberrant expression of miR-29c in uterine leiomyoma. Fertil Steril. 105:236–245 e1. 2016. View Article : Google Scholar : PubMed/NCBI
29	He B, Xiao YF, Tang B, Wu YY, Hu CJ, Xie R, Yang X, Yu ST, Dong H, Zhao XY, et al: hTERT mediates gastric cancer metastasis partially through the indirect targeting of ITGB1 by microRNA-29a. Sci Rep. 6:219552016. View Article : Google Scholar : PubMed/NCBI
30	Chen J, Gu L, Ni J, Hu P, Hu K and Shi YL: MiR-183 Regulates ITGB1P Expression and Promotes Invasion of Endometrial Stromal Cells. Biomed Res Int. 2015:3402182015.PubMed/NCBI
31	Jiang Z, Xu W, Dan G, Liu Y and Xiong J: P53 and murine double mimute 2 (MDM2) expression changes and significance in different types of endometrial lesions. Med Sci Monit. 22:4786–4793. 2016. View Article : Google Scholar : PubMed/NCBI
32	Bollaert E, de Rocca Serra A and Demoulin JB: The HMG box transcription factor HBP1: A cell cycle inhibitor at the crossroads of cancer signaling pathways. Cell Mol Life Sci. 76:1529–1539. 2019. View Article : Google Scholar : PubMed/NCBI
33	He S, Yang S, Niu M, Zhong Y, Dan Gao, Zhang Y, Ma H, Xiong W, Zhou M, Zhou Y, et al: HMG-box transcription factor 1: A positive regulator of the G1/S transition through the Cyclin-CDK-CDKI molecular network in nasopharyngeal carcinoma. Cell Death Dis. 9:1002018. View Article : Google Scholar : PubMed/NCBI
34	Conza D, Mirra P, Cali G, Tortora T, Insabato L, Fiory F, Schenone S, Amato R, Beguinot F, Perrotti N and Ulianich L: The SGK1 inhibitor SI113 induces autophagy, apoptosis, and endoplasmic reticulum stress in endometrial cancer cells. J Cell Physiol. 232:3735–3743. 2017. View Article : Google Scholar : PubMed/NCBI
35	Palmirotta R, Cives M, Della-Morte D, Capuani B, Lauro D, Guadagni F and Silvestris F: Sirtuins and cancer: role in the epithelial-mesenchymal transition. Oxid Med Cell Longev. 2016:30314592016. View Article : Google Scholar : PubMed/NCBI
36	Bartosch C, Monteiro-Reis S, Almeida-Rios D, Vieira R, Castro A, Moutinho M, Rodrigues M, Graça I, Lopes JM and Jerónimo C: Assessing sirtuin expression in endometrial carcinoma and non-neoplastic endometrium. Oncotarget. 7:1144–1154. 2016. View Article : Google Scholar : PubMed/NCBI
37	Asaka R, Miyamoto T, Yamada Y, Ando H, Mvunta DH, Kobara H and Shiozawa T: Sirtuin 1 promotes the growth and cisplatin resistance of endometrial carcinoma cells: A novel therapeutic target. Lab Invest. 95:1363–1373. 2015. View Article : Google Scholar : PubMed/NCBI
38	Bae HJ, Noh JH, Kim JK, Eun JW, Jung KH, Kim MG, Chang YG, Shen Q, Kim SJ, Park WS, et al: MicroRNA-29c functions as a tumor suppressor by direct targeting oncogenic SIRT1 in hepatocellular carcinoma. Oncogene. 33:2557–2567. 2014. View Article : Google Scholar : PubMed/NCBI
39	Mahecha AM and Wang H: The influence of vascular endothelial growth factor-A and matrix metalloproteinase-2 and −9 in angiogenesis, metastasis, and prognosis of endometrial cancer. Onco Targets Ther. 10:4617–4624. 2017. View Article : Google Scholar : PubMed/NCBI
40	Park SY, Lee JH, Ha M, Nam JW and Kim VN: miR-29 miRNAs activate p53 by targeting p85 alpha and CDC42. Nat Struct Mol Biol. 16:23–29. 2009. View Article : Google Scholar : PubMed/NCBI
41	Qadir MI, Parveen A and Ali M: Cdc42: Role in cancer management. Chem Biol Drug Des. 86:432–439. 2015. View Article : Google Scholar : PubMed/NCBI
42	Ye H, Zhang Y, Geng L and Li Z: Cdc42 expression in cervical cancer and its effects on cervical tumor invasion and migration. Int J Oncol. 46:757–763. 2015. View Article : Google Scholar : PubMed/NCBI
43	Feng Y, Fang S and Li M: Expression of P21-activated kinase 1 and cell division control protein 42 homolog correlates with clinicopathological features and prognosis in cervical carcinoma. J Obstet Gynaecol Res. 42:860–869. 2016. View Article : Google Scholar : PubMed/NCBI
44	Shi C, Ren L, Sun C, Yu L, Bian X, Zhou X, Wen Y, Hua D, Zhao S, Luo W, et al: miR-29a/b/c function as invasion suppressors for gliomas by targeting CDC42 and predict the prognosis of patients. Br J Cancer. 117:1036–1047. 2017. View Article : Google Scholar : PubMed/NCBI
45	Yang D, Zhang Y, Cheng Y, Hong L, Wang C, Wei Z, Cai Q and Yan R: High expression of cell division cycle 42 promotes pancratic cancer growth and predicts poor outcome of pancreatic cancer patients. Dig Dis Sci. 62:958–967. 2017. View Article : Google Scholar : PubMed/NCBI
46	Kamai T, Yamanishi T, Shirataki H, Takagi K, Asami H, Ito Y and Yoshida K: Overexpression of RhoA, Rac1 and Cdc42 GTPases is associated with progression in testicular cancer. Clin Cancer Res. 10:4799–4805. 2004. View Article : Google Scholar : PubMed/NCBI
47	Konno Y, Dong P, Xiong Y, Suzuki F, Lu J, Cai M, Watari H, Mitamura T, Hosaka M, Hanley SJ, et al: MicroRNA-101 targets EZH2, MCL-1 and FOS to suppress proliferation, invasion and stem cell-like phenotype of aggressive endometrial cancer cells. Oncotarget. 5:6049–6062. 2014. View Article : Google Scholar : PubMed/NCBI
48	Simmen F, Su Y, Xiao R, Zeng Z and Simmen RC: The Kruppel-like factor 9 (KLF9) network in HEC-1-A endometrial carcinoma cells suggests the carcinogenic potential of dys-regulated KLF9 expression. Reprod Biol Endocrinol. 6:412008. View Article : Google Scholar : PubMed/NCBI
49	Menkhorst E, Griffiths M, Van Sinderen M, Rainczuk K, Niven K and Dimitriadis E: Galectin-7 is elevated in endometrioid (type I) endometrial cancer and promotes cell migration. Oncol Lett. 16:4721–4728. 2018.PubMed/NCBI
50	Van Agtmael T and Bruckner-Tuderman L: Basement membranes and human disease. Cell Tissue Res. 339:167–188. 2010. View Article : Google Scholar : PubMed/NCBI
51	Kelly FD, Tawia SA and Rogers PA: Immunohistochemical characterization of human endometrial microvascular basement membrane components during the normal menstrual cycle. Hum Reprod. 10:268–276. 1995. View Article : Google Scholar : PubMed/NCBI