Assessment of Expression of Homeobox A5 in Endometrial Cancer on the mRNA and Protein Level

Author(s): Konrad Dziobek*, Marcin Oplawski, Nikola Zmarzły, Beniamin O. Gabarek, Robert Kiełbasiński, Kamil Kiełbasiński, Przemysław Kieszkowski, Karol Talkowski, Dariusz Boroń

Journal Name: Current Pharmaceutical Biotechnology

Volume 21 , Issue 7 , 2020


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Abstract:

Background: Endometrial cancer is one of the most common gynecological cancer in the developed countries and occurs mainly in postmenopausal women. Angiogenesis is important for cancer formation as it provides nutrients for growing tumor mass. Most tumors do not show detectable Homeobox A5 (HOXA5 level), suggesting its potential role as a cancer suppressor. It was demonstrated that HOXA5 is involved in the progression of various types of cancer and the loss of its expression correlates with higher pathological grade and poorer outcome.

Objective: The aim of the study was to evaluate HOXA5 expression at transcriptome and protein levels.

Materials and Methods: The study enrolled 45 women diagnosed with endometrial cancer and 15 without neoplastic changes. The histopathological examination allowed us to divide cancer tissue samples according to the degree of histological differentiation: G1, 17; G2, 15; G3, 13. The expression of the HOXA5 protein was determined by immunohistochemistry. Microarray and RT-qPCR techniques were used to assess HOXA5 expression at the mRNA level.

Results: The reaction to the HOXA5 protein was only visible in glandular cells in G1 endometrial cancer and was lower compared to the control. In grades 2 and 3, reactions were noted at the limit of the method’s sensitivity. In addition, reduced HOXA5 expression was observed at the transcriptome level.

Conclusion: HOXA5 may become a potential complementary molecular marker, allowing early detection of neoplastic changes in the endometrium. It also seems that detection of HOXA5 at the mRNA and protein levels may be helpful in improving the accuracy of diagnosis and planning effective oncological therapy.

Keywords: Endometrial cancer, HOXA5, immunohistochemistry, RT-qPCR, microarray, mRNA level.

[1]
Ryan, A.J.; Susil, B.; Jobling, T.W.; Oehler, M.K. Endometrial cancer. Cell Tissue Res., 2005, 322(1), 53-61.
[http://dx.doi.org/10.1007/s00441-005-1109-5] [PMID: 15947972]
[2]
Helpman, L.; Kupets, R.; Covens, A.; Saad, R.S.; Khalifa, M.A.; Ismiil, N.; Ghorab, Z.; Dubé, V.; Nofech-Mozes, S. Assessment of endometrial sampling as a predictor of final surgical pathology in endometrial cancer. Br. J. Cancer, 2014, 110(3), 609-615.
[http://dx.doi.org/10.1038/bjc.2013.766] [PMID: 24366295]
[3]
Liu, W.J.; Zhang, T.; Guo, Q.L.; Liu, C.Y.; Bai, Y.Q. Effect of ATRA on the expression of HOXA5 gene in K562 cells and its relationship with cell cycle and apoptosis. Mol. Med. Rep., 2016, 13(5), 4221-4228.
[http://dx.doi.org/10.3892/mmr.2016.5086] [PMID: 27052693]
[4]
Agger, K.; Cloos, P.A.; Christensen, J.; Pasini, D.; Rose, S.; Rappsilber, J.; Issaeva, I.; Canaani, E.; Salcini, A.E.; Helin, K. UTX and JMJD3 are histone H3K27 demethylases involved in HOX gene regulation and development. Nature, 2007, 449(7163), 731-734.
[http://dx.doi.org/10.1038/nature06145] [PMID: 17713478]
[5]
Liu, W.J.; Huang, M.X.; Guo, Q.L.; Chen, J.H.; Shi, H. Effect of human cytomegalovirus infection on the expression of Hoxb2 and Hoxb4 genes in the developmental process of cord blood erythroid progenitors. Mol. Med. Rep., 2011, 4(6), 1307-1311.
[http://dx.doi.org/10.3892/mmr.2011.576] [PMID: 21879259]
[6]
Bagadi, S.A.; Prasad, C.P.; Kaur, J.; Srivastava, A.; Prashad, R.; Gupta, S.D.; Ralhan, R. Clinical significance of promoter hypermethylation of RASSF1A, RARbeta2, BRCA1 and HOXA5 in breast cancers of Indian patients. Life Sci., 2008, 82(25-26), 1288-1292.
[http://dx.doi.org/10.1016/j.lfs.2008.04.020] [PMID: 18538349]
[7]
Huang, H.P.; Liu, W.J.; Guo, Q.L.; Bai, Y.Q. Effect of silencing HOXA5 gene expression using RNA interference on cell cycle and apoptosis in Jurkat cells. Int. J. Mol. Med., 2016, 37(3), 669-678.
[http://dx.doi.org/10.3892/ijmm.2016.2480] [PMID: 26846409]
[8]
Myers, C.; Charboneau, A.; Cheung, I.; Hanks, D.; Boudreau, N. Sustained expression of homeobox D10 inhibits angiogenesis. Am. J. Pathol., 2002, 161(6), 2099-2109.
[http://dx.doi.org/10.1016/S0002-9440(10)64488-4] [PMID: 12466126]
[9]
Chen, Y.Q.; Yang, T.Q.; Zhou, B.; Yang, M.X.; Feng, H.J.; Wang, Y.L. HOXA5 overexpression promotes osteosarcoma cell apoptosis through the p53 and p38α MAPK pathway. Gene, 2019, 689, 18-23.
[http://dx.doi.org/10.1016/j.gene.2018.11.081] [PMID: 30521886]
[10]
Cuevas, I.; Layman, H.; Coussens, L.; Boudreau, N. Sustained endothelial expression of HoxA5 in vivo impairs pathological angiogenesis and tumor progression. PLoS One, 2015, 10(3)e0121720
[http://dx.doi.org/10.1371/journal.pone.0121720] [PMID: 25821967]
[11]
Arderiu, G.; Cuevas, I.; Chen, A.; Carrio, M.; East, L.; Boudreau, N.J. HoxA5 stabilizes adherens junctions via increased Akt1. Cell Adhes. Migr., 2007, 1(4), 185-195.
[http://dx.doi.org/10.4161/cam.1.4.5448] [PMID: 19262140]
[12]
Zhang, M.L.; Nie, F.Q.; Sun, M.; Xia, R.; Xie, M.; Lu, K.H.; Li, W. HOXA5 indicates poor prognosis and suppresses cell proliferation by regulating p21 expression in non small cell lung cancer. Tumour Biol., 2015, 36(5), 3521-3531.
[http://dx.doi.org/10.1007/s13277-014-2988-4] [PMID: 25549794]
[13]
Jeannotte, L.; Gotti, F.; Landry-Truchon, K. Hoxa5, a key player in development and disease. J. Dev. Biol., 2016, 4(2), 13.
[http://dx.doi.org/10.3390/jdb4020013] [PMID: 29615582]
[14]
Wang, C.C.; Su, K.Y.; Chen, H.Y.; Chang, S.Y.; Shen, C.F.; Hsieh, C.H.; Hong, Q.S.; Chiang, C.C.; Chang, G.C.; Yu, S.L.; Chen, J.J. HOXA5 inhibits metastasis via regulating cytoskeletal remodelling and associates with prolonged survival in non-small-cell lung carcinoma. PLoS One, 2015, 10(4)e0124191
[http://dx.doi.org/10.1371/journal.pone.0124191] [PMID: 25875824]
[15]
Opławski, M.; Michalski, M.; Witek, A.; Michalski, B.; Zmarzły, N.; Jęda-Golonka, A.; Styblińska, M.; Gola, J.; Kasprzyk-Żyszczyńska, M.; Mazurek, U.; Plewka, A. Identification of a gene expression profile associated with the regulation of angiogenesis in endometrial cancer. Mol. Med. Rep., 2017, 16(3), 2547-2555.
[http://dx.doi.org/10.3892/mmr.2017.6868] [PMID: 28656251]
[16]
Abdalla, A.M.E.; Xiao, L.; Ullah, M.W.; Yu, M.; Ouyang, C.; Yang, G. Current challenges of cancer anti-angiogenic therapy and the promise of nanotherapeutics. Theranostics, 2018, 8(2), 533-548.
[http://dx.doi.org/10.7150/thno.21674] [PMID: 29290825]
[17]
Teo, W.W.; Merino, V.F.; Cho, S.; Korangath, P.; Liang, X.; Wu, R.C.; Neumann, N.M.; Ewald, A.J.; Sukumar, S. HOXA5 determines cell fate transition and impedes tumor initiation and progression in breast cancer through regulation of E-cadherin and CD24. Oncogene, 2016, 35(42), 5539-5551.
[http://dx.doi.org/10.1038/onc.2016.95] [PMID: 27157614]
[18]
Chen, H.; Chung, S.; Sukumar, S. HOXA5-induced apoptosis in breast cancer cells is mediated by caspases 2 and 8. Mol. Cell. Biol., 2004, 24(2), 924-935.
[http://dx.doi.org/10.1128/MCB.24.2.924-935.2004] [PMID: 14701762]
[19]
Raman, V.; Martensen, S.A.; Reisman, D.; Evron, E.; Odenwald, W.F.; Jaffee, E.; Marks, J.; Sukumar, S. Compromised HOXA5 function can limit p53 expression in human breast tumours. Nature, 2000, 405(6789), 974-978.
[http://dx.doi.org/10.1038/35016125] [PMID: 10879542]
[20]
Rodini, C.O.; Xavier, F.C.; Paiva, K.B.; De Souza Setúbal Destro, M.F.; Moyses, R.A.; Michaluarte, P.; Carvalho, M.B.; Fukuyama, E.E.; Tajara, E.H.; Okamoto, O.K.; Nunes, F.D. Head And Neck Genome Project Gencapo. Homeobox gene expression profile indicates HOXA5 as a candidate prognostic marker in oral squamous cell carcinoma. Int. J. Oncol., 2012, 40(4), 1180-1188.
[http://dx.doi.org/10.3892/ijo.2011.1321] [PMID: 22227861]
[21]
Tan, S.H.; Barker, N. Stemming colorectal cancer growth and metastasis, HOXA5 forces cancer stem cells to differentiate. Cancer Cell, 2015, 28(6), 683-685.
[http://dx.doi.org/10.1016/j.ccell.2015.11.004] [PMID: 26678334]
[22]
Amant, F.; Moerman, P.; Neven, P.; Timmerman, D.; Van Limbergen, E.; Vergote, I. Endometrial cancer. Lancet, 2005, 366(9484), 491-505.
[http://dx.doi.org/10.1016/S0140-6736(05)67063-8] [PMID: 16084259]
[23]
Vilarrasa-Blasi, R.; Soler-Vila, P.; Verdaguer-Dot, N.; Russiñol, N.; Di Stefano, M.; Chapaprieta, V.; Prosper, F. Dynamics of genome architecture and chromatin function during human B cell differentiation and neoplastic transformation. bioRxiv, 2019, •••764910
[24]
Ordóñez-Morán, P.; Dafflon, C.; Imajo, M.; Nishida, E.; Huelsken, J. HOXA5 counteracts stem cell traits by inhibiting Wnt signaling in colorectal cancer. Cancer Cell, 2015, 28(6), 815-829.
[http://dx.doi.org/10.1016/j.ccell.2015.11.001] [PMID: 26678341]
[25]
Chatterjee, A.; Rodger, E.J.; Eccles, M.R. Epigenetic drivers of tumourigenesis and cancer metastasis. Semin. Cancer Biol., 2018, 51, 149-159.
[http://dx.doi.org/10.1016/j.semcancer.2017.08.004] [PMID: 28807546]
[26]
Pidsley, R.; Lawrence, M.G.; Zotenko, E.; Niranjan, B.; Statham, A.; Song, J.; Chabanon, R.M.; Qu, W.; Wang, H.; Richards, M.; Nair, S.S.; Armstrong, N.J.; Nim, H.T.; Papargiris, M.; Balanathan, P.; French, H.; Peters, T.; Norden, S.; Ryan, A.; Pedersen, J.; Kench, J.; Daly, R.J.; Horvath, L.G.; Stricker, P.; Frydenberg, M.; Taylor, R.A.; Stirzaker, C.; Risbridger, G.P.; Clark, S.J. Enduring epigenetic landmarks define the cancer microenvironment. Genome Res., 2018, 28(5), 625-638.
[http://dx.doi.org/10.1101/gr.229070.117] [PMID: 29650553]
[27]
Watson, R.E.; Curtin, G.M.; Hellmann, G.M.; Doolittle, D.J.; Goodman, J.I. Increased DNA methylation in the HoxA5 promoter region correlates with decreased expression of the gene during tumor promotion. Mol. Carcinog., 2004, 41(1), 54-66.
[http://dx.doi.org/10.1002/mc.20043] [PMID: 15352125]
[28]
Li, D.; Bai, Y.; Feng, Z.; Li, W.; Yang, C.; Guo, Y.; Lin, C.; Zhang, Y.; He, Q.; Hu, G.; Li, X. Study of promoter methylation patterns of HOXA2, HOXA5 and HOXA6 and its clinicopathological characteristics in colorectal cancer. Front. Oncol., 2019, 9, 394.
[http://dx.doi.org/10.3389/fonc.2019.00394] [PMID: 31165042]
[29]
Issa, J.P. Cancer prevention: epigenetics steps up to the plate. Cancer Prev. Res. (Phila.), 2008, 1(4), 219-222.
[http://dx.doi.org/10.1158/1940-6207.CAPR-08-0029] [PMID: 19138962]
[30]
Wang, Z.; Yu, C.; Wang, H. HOXA5 inhibits the proliferation and induces the apoptosis of cervical cancer cells via regulation of protein kinase B and p27. Oncol. Rep., 2019, 41(2), 1122-1130.
[PMID: 30483748]
[31]
Rhoads, K.; Arderiu, G.; Charboneau, A.; Hansen, S.L.; Hoffman, W.; Boudreau, N. A role for Hox A5 in regulating angiogenesis and vascular patterning. Lymphat. Res. Biol., 2005, 3(4), 240-252.
[http://dx.doi.org/10.1089/lrb.2005.3.240] [PMID: 16379594]
[32]
Romero-Cordoba, S.L.; Salido-Guadarrama, I.; Rodriguez-Dorantes, M.; Hidalgo-Miranda, A. miRNA biogenesis: biological impact in the development of cancer. Cancer Biol. Ther., 2014, 15(11), 1444-1455.
[http://dx.doi.org/10.4161/15384047.2014.955442] [PMID: 25482951]
[33]
Hukowska-Szematowicz, B.; Deptuła, B. Biologiczna rola mikroRNA (miRNA) – nowe dane. Post Biol Kom., 2010, 37, 1-13.
[34]
Kavitha, N.; Vijayarathna, S.; Jothy, S.L.; Oon, C.E.; Chen, Y.; Kanwar, J.R.; Sasidharan, S. MicroRNAs: biogenesis, roles for carcinogenesis and as potential biomarkers for cancer diagnosis and prognosis. Asian Pac. J. Cancer Prev., 2014, 15(18), 7489-7497.
[http://dx.doi.org/10.7314/APJCP.2014.15.18.7489] [PMID: 25292018]
[35]
Liu, X.H.; Lu, K.H.; Wang, K.M.; Sun, M.; Zhang, E.B.; Yang, J.S.; Yin, D.D.; Liu, Z.L.; Zhou, J.; Liu, Z.J.; De, W.; Wang, Z.X. MicroRNA-196a promotes non-small cell lung cancer cell proliferation and invasion through targeting HOXA5. BMC Cancer, 2012, 12, 348.
[http://dx.doi.org/10.1186/1471-2407-12-348] [PMID: 22876840]
[36]
Wcisło-Dziadecka, D.; Grabarek, B.; Zmarzły, N.; Skubis, A.; Sikora, B.; Kruszniewska-Rajs, C.; Gola, J.; Mazurek, U.; Kucharz, E. Influence of adalimumab on the expression profile of genes associated with the histaminergic system in the skin fibroblast in vitro. BioMed Res. Int., 2018, 20181582173
[http://dx.doi.org/10.1155/2018/1582173] [PMID: 29487864]
[37]
Rudini, N.; Novello, C.; Destro, A.; Riboldi, E.; Donadon, M.; Viganò, L.; Morenghi, E.; Roncalli, M.; Di Tommaso, L. Phenotypic and molecular changes in nodule-in-nodule hepatocellular carcinoma with pathogenetic implications. Histopathology, 2018, 73(4), 601-611.
[http://dx.doi.org/10.1111/his.13659] [PMID: 29791027]
[38]
Cohen, S.A.; Pritchard, C.C.; Jarvik, G.P. Lynch syndrome: From screening to diagnosis to treatment in the era of modern molecular oncology. Annu. Rev. Genomics Hum. Genet., 2019, 20, 293-307.
[http://dx.doi.org/10.1146/annurev-genom-083118-015406] [PMID: 30848956]
[39]
Oplawski, M.; Dziobek, K.; Zmarzły, N.; Grabarek, B.O.; Kiełbasiński, R.; Kieszkowski, P.; Januszyk, P.; Talkowski, K.; Plewka, A.; Boroń, D. Variances in the level of of COX-2 and iNOS in different grades of endometrial cancer. Curr. Pharm. Biotechnol., 2020, 21(1), 52-59.
[http://dx.doi.org/10.2174/1389201020666190918104105] [PMID: 31533599]


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Article Details

VOLUME: 21
ISSUE: 7
Year: 2020
Published on: 27 December, 2019
Page: [635 - 641]
Pages: 7
DOI: 10.2174/1389201021666191227121627

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