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Knockdown of MPP8 suppresses cell proliferation via regulation of HOXA5 in non-small cell lung cancer cells
Corresponding Author(s) : Xiang-Yang Gao
Cellular and Molecular Biology,
Vol. 64 No. 1: Issue 1
Abstract
M-phase phosphoprotein 8 (MPP8) is reported to be closely implicated in cancer initiation and progression. In addition, the homeobox gene HOXA5 has been shown to play critical roles in hematopoiesis, embryogenesis, and tumorigenesis. Nevertheless, the functional relevance of MPP8 and it's relation with HOXA5 in non-small cell lung cancer (NSCLC) is unknown. Therefore, the present study aimed to detect the expression profile of MPP8 in NSCLC and further explore it's biological roles in lung cancer cells. Cell proliferation was measured by CCK-8 assay and EdU incorporation assay. Real-time PCR was applied to detect the mRNA expression of MPP8 and HOXA5. The protein levels of MPP8 and HOXA5 were evaluated by western blot. Our study found that the expression of MPP8 was significantly increased in the NSCLC tissue compared with the adjacent non-tumorous tissue. Compared with the human lung fibroblasts, the elevated gene expression of MPP8 was also detected in the human NSCLC cell lines including NCI-H23 and NCI-H1299. In addition, knockdown of MPP8 led to an obvious reduction in cell viability and DNA synthesis in NCI-H23 and NCI-H1299 cells. Furthermore, down-regulation of MPP8 resulted in elevated expression of HOXA5 in NSCLC cells both at the mRNA and protein levels. Moreover, depletion of HOXA5 abolished the anti-tumor function of MPP8 knockdown in NSCLC cells. The present study demonstrated that MPP8 was associated with NSCLC cell proliferation through regulation of HOXA5, suggesting that MPP8 may act as a novel therapeutic target for treatment of NSCLC.
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- References
- Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin 2016; 66:7-30.
- Brambilla E, Le Teuff G, Marguet S, Lantuejoul S, Dunant A, Graziano S, et al. Prognostic effect of tumor lymphocytic infiltration in resectable Non-Small-Cell lung cancer. J Clin Oncol 2016; 34:1223-1230.
- Kumarakulasinghe NB, van Zanwijk N, Soo RA. Molecular targeted therapy in the treatment of advanced stage non-small cell lung cancer (NSCLC). Respirology 2015; 20:370-378.
- Ileana EE, Wistuba II, Izzo JG. From uniplex to multiplex molecular profiling in advanced Non-Small cell lung carcinoma. Cancer J 2015; 21:413-424.
- Matsumoto-Taniura N, Pirollet F, Monroe R, Gerace L, Westendorf JM. Identification of novel M phase phosphoproteins by expression cloning. Mol Biol Cell 1996; 7:1455-1469.
- Chang Y, Sun L, Kokura K, Horton JR, Fukuda M, Espejo A, et al. MPP8 mediates the interactions between DNA methyltransferase Dnmt3a and H3K9 methyltransferase GLP/G9a. Nat Commun 2011; 2:533.
- Nishigaki M, Kawada Y, Misaki T, Murata K, Goshima T, Hirokawa T, et al. Mitotic phosphorylation of MPP8 by cyclin-dependent kinases regulates chromatin dissociation. Biochem Biophys Res Commun 2013; 432:654-659.
- Kokura K, Sun L, Bedford MT, Fang J. Methyl-H3K9-binding protein MPP8 mediates E-cadherin gene silencing and promotes tumour cell motility and invasion. EMBO J 2010; 29:3673-3687.
- Maximov VV, Martynenko AV, Arman IP, Tarantul VZ. Humanin binds MPP8: Mapping interaction sites of the peptide and protein. J Pept Sci 2013; 19:301-307.
- Plowright L, Harrington KJ, Pandha HS, Morgan R. HOX transcription factors are potential therapeutic targets in non-small-cell lung cancer (targeting HOX genes in lung cancer). Br J Cancer 2009; 100:470-475.
- Ordonez-Moran P, Dafflon C, Imajo M, Nishida E, Huelsken J. HOXA5 counteracts stem cell traits by inhibiting wnt signaling in colorectal cancer. Cancer Cell 2015; 28:815-829.
- Teo WW, Merino VF, Cho S, Korangath P, Liang X, Wu RC, et al. HOXA5 determines cell fate transition and impedes tumor initiation and progression in breast cancer through regulation of E-cadherin and CD24. Oncogene 2016; 35:5539-5551.
- Yoo KH, Park YK, Kim HS, Jung WW, Chang SG. Epigenetic inactivation of HOXA5 and MSH2 gene in clear cell renal cell carcinoma. Pathol Int 2010; 60:661-666.
- Nunes FD, de Almeida FC, Tucci R, de Sousa SC. Homeobox genes: A molecular link between development and cancer. Pesqui Odontol Bras 2003; 17:94-98.
- Abate-Shen C. Deregulated homeobox gene expression in cancer: Cause or consequence? Nat Rev Cancer 2002; 2:777-785.
- Grier DG, Thompson A, Kwasniewska A, McGonigle GJ, Halliday HL, Lappin TR. The pathophysiology of HOX genes and their role in cancer. J Pathol 2005; 205:154-171.
- You JS, Jones PA. Cancer genetics and epigenetics: Two sides of the same coin? Cancer Cell 2012; 22:9-20.
- Sadikovic B, Al-Romaih K, Squire JA, Zielenska M. Cause and consequences of genetic and epigenetic alterations in human cancer. Curr Genomics 2008; 9:394-408.
- Liang X, Liu T, Zhang W, Zhang K, Guo S, Liang J. Lentivirus-mediated knockdown of M-phase phosphoprotein 8 inhibits proliferationNof colon cancer cells. Biotechnol Appl Biochem 2016.
- Iimura T, Pourquie O. Hox genes in time and space during vertebrate body formation. Dev Growth Differ 2007; 49:265-275.
- Abramovich C, Humphries RK. Hox regulation of normal and leukemic hematopoietic stem cells. Curr Opin Hematol 2005; 12:210-216.
- Daniels TR, Neacato II, Rodriguez JA, Pandha HS, Morgan R, Penichet ML. Disruption of HOX activity leads to cell death that can be enhanced by the interference of iron uptake in malignant B cells. Leukemia 2010; 24:1555-1565.
- Tan SH, Barker N. Stemming colorectal cancer growth and metastasis: HOXA5 forces cancer stem cells to differentiate. Cancer Cell 2015; 28:683-685.
- Wang CC, Su KY, Chen HY, Chang SY, Shen CF, Hsieh CH, et al. HOXA5 inhibits metastasis via regulating cytoskeletal remodelling and associates with prolonged survival in non-small-cell lung carcinoma. PLoS One 2015; 10:e124191.
- Mandeville I, Aubin J, LeBlanc M, Lalancette-Hebert M, Janelle MF, Tremblay GM, et al. Impact of the loss of Hoxa5 function on lung alveogenesis. Am J Pathol 2006; 169:1312-1327.
References
References
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin 2016; 66:7-30.
Brambilla E, Le Teuff G, Marguet S, Lantuejoul S, Dunant A, Graziano S, et al. Prognostic effect of tumor lymphocytic infiltration in resectable Non-Small-Cell lung cancer. J Clin Oncol 2016; 34:1223-1230.
Kumarakulasinghe NB, van Zanwijk N, Soo RA. Molecular targeted therapy in the treatment of advanced stage non-small cell lung cancer (NSCLC). Respirology 2015; 20:370-378.
Ileana EE, Wistuba II, Izzo JG. From uniplex to multiplex molecular profiling in advanced Non-Small cell lung carcinoma. Cancer J 2015; 21:413-424.
Matsumoto-Taniura N, Pirollet F, Monroe R, Gerace L, Westendorf JM. Identification of novel M phase phosphoproteins by expression cloning. Mol Biol Cell 1996; 7:1455-1469.
Chang Y, Sun L, Kokura K, Horton JR, Fukuda M, Espejo A, et al. MPP8 mediates the interactions between DNA methyltransferase Dnmt3a and H3K9 methyltransferase GLP/G9a. Nat Commun 2011; 2:533.
Nishigaki M, Kawada Y, Misaki T, Murata K, Goshima T, Hirokawa T, et al. Mitotic phosphorylation of MPP8 by cyclin-dependent kinases regulates chromatin dissociation. Biochem Biophys Res Commun 2013; 432:654-659.
Kokura K, Sun L, Bedford MT, Fang J. Methyl-H3K9-binding protein MPP8 mediates E-cadherin gene silencing and promotes tumour cell motility and invasion. EMBO J 2010; 29:3673-3687.
Maximov VV, Martynenko AV, Arman IP, Tarantul VZ. Humanin binds MPP8: Mapping interaction sites of the peptide and protein. J Pept Sci 2013; 19:301-307.
Plowright L, Harrington KJ, Pandha HS, Morgan R. HOX transcription factors are potential therapeutic targets in non-small-cell lung cancer (targeting HOX genes in lung cancer). Br J Cancer 2009; 100:470-475.
Ordonez-Moran P, Dafflon C, Imajo M, Nishida E, Huelsken J. HOXA5 counteracts stem cell traits by inhibiting wnt signaling in colorectal cancer. Cancer Cell 2015; 28:815-829.
Teo WW, Merino VF, Cho S, Korangath P, Liang X, Wu RC, et al. HOXA5 determines cell fate transition and impedes tumor initiation and progression in breast cancer through regulation of E-cadherin and CD24. Oncogene 2016; 35:5539-5551.
Yoo KH, Park YK, Kim HS, Jung WW, Chang SG. Epigenetic inactivation of HOXA5 and MSH2 gene in clear cell renal cell carcinoma. Pathol Int 2010; 60:661-666.
Nunes FD, de Almeida FC, Tucci R, de Sousa SC. Homeobox genes: A molecular link between development and cancer. Pesqui Odontol Bras 2003; 17:94-98.
Abate-Shen C. Deregulated homeobox gene expression in cancer: Cause or consequence? Nat Rev Cancer 2002; 2:777-785.
Grier DG, Thompson A, Kwasniewska A, McGonigle GJ, Halliday HL, Lappin TR. The pathophysiology of HOX genes and their role in cancer. J Pathol 2005; 205:154-171.
You JS, Jones PA. Cancer genetics and epigenetics: Two sides of the same coin? Cancer Cell 2012; 22:9-20.
Sadikovic B, Al-Romaih K, Squire JA, Zielenska M. Cause and consequences of genetic and epigenetic alterations in human cancer. Curr Genomics 2008; 9:394-408.
Liang X, Liu T, Zhang W, Zhang K, Guo S, Liang J. Lentivirus-mediated knockdown of M-phase phosphoprotein 8 inhibits proliferationNof colon cancer cells. Biotechnol Appl Biochem 2016.
Iimura T, Pourquie O. Hox genes in time and space during vertebrate body formation. Dev Growth Differ 2007; 49:265-275.
Abramovich C, Humphries RK. Hox regulation of normal and leukemic hematopoietic stem cells. Curr Opin Hematol 2005; 12:210-216.
Daniels TR, Neacato II, Rodriguez JA, Pandha HS, Morgan R, Penichet ML. Disruption of HOX activity leads to cell death that can be enhanced by the interference of iron uptake in malignant B cells. Leukemia 2010; 24:1555-1565.
Tan SH, Barker N. Stemming colorectal cancer growth and metastasis: HOXA5 forces cancer stem cells to differentiate. Cancer Cell 2015; 28:683-685.
Wang CC, Su KY, Chen HY, Chang SY, Shen CF, Hsieh CH, et al. HOXA5 inhibits metastasis via regulating cytoskeletal remodelling and associates with prolonged survival in non-small-cell lung carcinoma. PLoS One 2015; 10:e124191.
Mandeville I, Aubin J, LeBlanc M, Lalancette-Hebert M, Janelle MF, Tremblay GM, et al. Impact of the loss of Hoxa5 function on lung alveogenesis. Am J Pathol 2006; 169:1312-1327.