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01-04-2017 | Original Paper

HPIP promotes epithelial-mesenchymal transition and cisplatin resistance in ovarian cancer cells through PI3K/AKT pathway activation

Authors: Suresh Bugide, Vijay Kumar Gonugunta, Vasudevarao Penugurti, Vijaya Lakshmi Malisetty, Ratna K. Vadlamudi, Bramanandam Manavathi

Published in: Cellular Oncology | Issue 2/2017

Abstract

Purpose

Hematopoietic PBX interacting protein (HPIP), a scaffold protein, is known to regulate the proliferation, migration and invasion in different cancer cell types. The aim of this study was to assess the role of HPIP in ovarian cancer cell migration, invasion and epithelial-mesenchymal transition (EMT), and to unravel the mechanism by which it regulates these processes.

Methods

HPIP expression was assessed by immunohistochemistry of tissue microarrays containing primary ovarian tumor samples of different grades. OAW42, an ovarian carcinoma-derived cell line exhibiting a high HPIP expression, was used to study the role of HPIP in cell migration, invasion and EMT. HPIP knockdown in these cells was achieved using a small hairpin RNA (shRNA) approach. Cell migration and invasion were assessed using scratch wound and transwell invasion assays, respectively. The extent of EMT was assessed by determining the expression levels of Snail, Vimentin and E-cadherin using Western blotting. The effect of HPIP expression on AKT and MAPK activation was also investigated by Western blotting. Cell viabilities in response to cisplatin treatment were assessed using a MTT assay, whereas apoptosis was assessed by determining caspase-3 and PARP cleavage in ovarian carcinoma-derived SKOV3 cells.

Results

We found that HPIP is highly expressed in high-grade primary ovarian tumors. In addition, we found that HPIP promotes the migration, invasion and EMT in OAW42 cells and induces EMT in these cells via activation of the PI3K/AKT pathway. The latter was found to lead to stabilization of the Snail protein and to repression of E-cadherin expression through inactivation of GSK-3β. We also found that HPIP expression confers cisplatin resistance to SKOV3 cells after prolonged exposure and that its subsequent knockdown decreases the viability of these cells and increases caspase-3 activation and PARP proteolysis in these cells following cisplatin treatment.

Conclusions

From these results we conclude that HPIP expression is associated with high-grade ovarian tumors and may promote their migration, invasion and EMT, a process that is associated with metastasis. In addition, we conclude that HPIP may serve as a potential therapeutic target for cisplatin resistant ovarian tumors.

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R.P. Perez, A.K. Godwin, T.C. Hamilton, R.F. Ozols, Ovarian cancer biology. Semin Oncol 18, 186–204 (1991)PubMed

K.R. Cho, M. Shih Ie, Ovarian cancer. Ann Rev Pathol 4, 287–313 (2009). doi:10.1146/annurev.pathol.4.110807.092246 CrossRef

T.A. Yap, C.P. Carden, S.B. Kaye, Beyond chemotherapy: targeted therapies in ovarian cancer. Nat Rev Cancer 9, 167–181 (2009). doi:10.1038/nrc2583 CrossRefPubMed

N. Colombo, T. Van Gorp, G. Parma, F. Amant, G. Gatta, C. Sessa, I. Vergote, Ovarian cancer. Crit Rev Oncol Hematol 60, 159–179 (2006). doi:10.1016/j.critrevonc.2006.03.004 CrossRefPubMed

S. Patel, L. Kumar, N. Singh, Metformin and epithelial ovarian cancer therapeutics. Cell Oncol 38, 365–375 (2015). doi:10.1007/s13402-015-0235-7 CrossRef

M. Zou, X. Zhang, C. Xu, IL6-induced metastasis modulators p-STAT3, MMP-2 and MMP-9 are targets of 3,3'-diindolylmethane in ovarian cancer cells. Cell Oncol 39, 47–57 (2016). doi:10.1007/s13402-015-0251-7 CrossRef

R.C. Bast Jr., B. Hennessy, G.B. Mills, The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer 9, 415–428 (2009). doi:10.1038/nrc2644 CrossRefPubMedPubMedCentral

H. Naora, D.J. Montell, Ovarian cancer metastasis: integrating insights from disparate model organisms. Nat Rev Cancer 5, 355–366 (2005). doi:10.1038/nrc1611 CrossRefPubMed

J. Mikula-Pietrasik, P. Uruski, K. Matuszkiewicz, S. Szubert, R. Moszynski, D. Szpurek, S. Sajdak, A. Tykarski, K. Ksiazek, Ovarian cancer-derived ascitic fluids induce a senescence-dependent pro-cancerogenic phenotype in normal peritoneal mesothelial cells. Cell Oncol 39, 473–481 (2016). doi:10.1007/s13402-016-0289-1 CrossRef

10.

Y. Kuwabara, T. Yamada, K. Yamazaki, W.L. Du, K. Banno, D. Aoki, M. Sakamoto, Establishment of an ovarian metastasis model and possible involvement of E-cadherin down-regulation in the metastasis. Cancer Sci 99, 1933–1939 (2008). doi:10.1111/j.1349-7006.2008.00946.x PubMed

11.

D. Vergara, B. Merlot, J.P. Lucot, P. Collinet, D. Vinatier, I. Fournier, M. Salzet, Epithelial-mesenchymal transition in ovarian cancer. Cancer Lett 291, 59–66 (2010). doi:10.1016/j.canlet.2009.09.017 CrossRefPubMed

12.

B.T. Hennessy, D.L. Smith, P.T. Ram, Y. Lu, G.B. Mills, Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat Rev Drug Discov 4, 988–1004 (2005). doi:10.1038/nrd1902 CrossRefPubMed

13.

I. Vivanco, C.L. Sawyers, The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer 2, 489–501 (2002). doi:10.1038/nrc839 CrossRefPubMed

14.

B. Manavathi, F. Acconcia, S.K. Rayala, R. Kumar, An inherent role of microtubule network in the action of nuclear receptor. Proc Natl Acad Sci U S A 103, 15981–15986 (2006). doi:10.1073/pnas.0607445103 CrossRefPubMedPubMedCentral

15.

X. Xu, Z. Fan, L. Kang, J. Han, C. Jiang, X. Zheng, Z. Zhu, H. Jiao, J. Lin, K. Jiang, L. Ding, H. Zhang, L. Cheng, H. Fu, Y. Song, Y. Jiang, J. Liu, R. Wang, N. Du, Q. Ye, Hepatitis B virus X protein represses miRNA-148a to enhance tumorigenesis. J Clin Invest 123, 630–645 (2013). doi:10.1172/JCI64265 PubMedPubMedCentral

16.

S. Bugide, D. David, A. Nair, N. Kannan, V.S. Samanthapudi, J. Prabhakar, B. Manavathi, Hematopoietic PBX-interacting protein (HPIP) is over expressed in breast infiltrative ductal carcinoma and regulates cell adhesion and migration through modulation of focal adhesion dynamics. Oncogene 34, 4601–4612 (2015). doi:10.1038/onc.2014.389 CrossRefPubMed

17.

Y. Feng, X. Xu, Y. Zhang, J. Ding, Y. Wang, X. Zhang, Z. Wu, L. Kang, Y. Liang, L. Zhou, S. Song, K. Zhao, Q. Ye, HPIP is upregulated in colorectal cancer and regulates colorectal cancer cell proliferation, apoptosis and invasion. Sci Rep 5, 9429 (2015). doi:10.1038/srep09429 CrossRefPubMedPubMedCentral

18.

Y. Feng, L. Li, X. Zhang, Y. Zhang, Y. Liang, J. Lv, Z. Fan, J. Guo, T. Hong, B. Ji, Q. Ji, G. Mei, L. Ding, S. Zhang, X. Xu, Q. Ye, HPIP is overexpressed in gastric cancer and promotes gastric cancer cell proliferation, migration and invasion. Cancer Sci 106, 1313–1322 (2015). doi:10.1111/cas.12754 CrossRefPubMedPubMedCentral

19.

J. Pan, Y. Qin, M. Zhang, HPIP promotes non-small cell lung cancer cell proliferation, migration and invasion through regulation of the sonic hedgehog signaling pathway. Biomed Pharmacother 77, 176–181 (2016). doi:10.1016/j.biopha.2015.12.012 CrossRefPubMed

20.

S. Okada, T. Irie, J. Tanaka, R. Yasuhara, G. Yamamoto, T. Isobe, C. Hokazono, T. Tachikawa, Y. Kohno, K. Mishima, Potential role of hematopoietic pre-B-cell leukemia transcription factor-interacting protein in oral carcinogenesis. J Oral Pathol Med 44, 115–125 (2015). doi:10.1111/jop.12210 CrossRefPubMed

21.

S.C. Wang, D.S. Chai, C.B. Chen, Z.Y. Wang, L. Wang, HPIP promotes thyroid cancer cell growth, migration and EMT through activating PI3K/AKT signaling pathway. Biomed Pharmacother 75, 33–39 (2015). doi:10.1016/j.biopha.2015.08.027 CrossRefPubMed

22.

D.G. van Vuurden, E. Aronica, E. Hulleman, L.E. Wedekind, D. Biesmans, A. Malekzadeh, M. Bugiani, D. Geerts, D.P. Noske, W.P. Vandertop, G.J. Kaspers, J. Cloos, T. Wurdinger, P.P. van der Stoop, Pre-B-cell leukemia homeobox interacting protein 1 is overexpressed in astrocytoma and promotes tumor cell growth and migration. Neuro-Oncology 16, 946–959 (2014). doi:10.1093/neuonc/not308 CrossRefPubMedPubMedCentral

23.

K. Shostak, F. Patrascu, S.I. Goktuna, P. Close, L. Borgs, L. Nguyen, F. Olivier, A. Rammal, H. Brinkhaus, M. Bentires-Alj, J.C. Marine, A. Chariot, MDM2 restrains estrogen-mediated AKT activation by promoting TBK1-dependent HPIP degradation. Cell Death Differ 21, 811–824 (2014). doi:10.1038/cdd.2014.2 CrossRefPubMedPubMedCentral

24.

V.N. Gajulapalli, V.S. Samanthapudi, M. Pulaganti, S.S. Khumukcham, V.L. Malisetty, L. Guruprasad, S.K. Chitta, B. Manavathi, A transcriptional repressive role for epithelial-specific ETS factor ELF3 on oestrogen receptor alpha in breast cancer cells. Biochem J 473, 1047–1061 (2016). doi:10.1042/BCJ20160019 CrossRefPubMed

25.

G.R. Sareddy, B.C. Nair, S.K. Krishnan, V.K. Gonugunta, Q.G. Zhang, T. Suzuki, N. Miyata, A.J. Brenner, D.W. Brann, R.K. Vadlamudi, KDM1 is a novel therapeutic target for the treatment of gliomas. Oncotarget 4, 18–28 (2013). doi:10.18632/oncotarget.725 PubMed

26.

T. Bonome, D.A. Levine, J. Shih, M. Randonovich, C.A. Pise-Masison, F. Bogomolniy, L. Ozbun, J. Brady, J.C. Barrett, J. Boyd, M.J. Birrer, A gene signature predicting for survival in suboptimally debulked patients with ovarian cancer. Cancer Res 68, 5478–5486 (2008). doi:10.1158/0008-5472.CAN-07-6595 CrossRefPubMed

27.

R. Kalluri, R.A. Weinberg, The basics of epithelial-mesenchymal transition. J Clin Invest 119, 1420–1428 (2009). doi:10.1172/JCI39104 CrossRefPubMedPubMedCentral

28.

C.L. Chaffer, R.A. Weinberg, A perspective on cancer cell metastasis. Science 331, 1559–1564 (2011). doi:10.1126/science.1203543 CrossRefPubMed

29.

E. Batlle, E. Sancho, C. Franci, D. Dominguez, M. Monfar, J. Baulida, A. Garcia De Herreros, The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol 2, 84–89 (2000). doi:10.1038/35000034 CrossRefPubMed

30.

J.P. Thiery, H. Acloque, R.Y. Huang, M.A. Nieto, Epithelial-mesenchymal transitions in development and disease. Cell 139, 871–890 (2009). doi:10.1016/j.cell.2009.11.007 CrossRefPubMed

31.

B.P. Zhou, J. Deng, W. Xia, J. Xu, Y.M. Li, M. Gunduz, M.C. Hung, Dual regulation of snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat Cell Biol 6, 931–940 (2004). doi:10.1038/ncb1173 CrossRefPubMed

32.

D.A. Cross, D.R. Alessi, P. Cohen, M. Andjelkovich, B.A. Hemmings, Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 378, 785–789 (1995). doi:10.1038/378785a0 CrossRefPubMed

33.

A. Cano, M.A. Perez-Moreno, I. Rodrigo, A. Locascio, M.J. Blanco, M.G. del Barrio, F. Portillo, M.A. Nieto, The transcription factor snail controls epithelial-mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2, 76–83 (2000). doi:10.1038/35000025 CrossRefPubMed

34.

S. Lee, E.J. Choi, C. Jin, D.H. Kim, Activation of PI3K/Akt pathway by PTEN reduction and PIK3CA mRNA amplification contributes to cisplatin resistance in an ovarian cancer cell line. Gynecol Oncol 97, 26–34 (2005). doi:10.1016/j.ygyno.2004.11.051 CrossRefPubMed

35.

H. Asechi, E. Hatano, T. Nitta, M. Tada, K. Iwaisako, N. Tamaki, H. Nagata, M. Narita, A. Yanagida, I. Ikai, S. Uemoto, Resistance to cisplatin-induced apoptosis via PI3K-dependent survivin expression in a rat hepatoma cell line. Int J Oncol 37, 89–96 (2010)PubMed

36.

E. Lengyel, Ovarian cancer development and metastasis. Am J Pathol 177, 1053–1064 (2010). doi:10.2353/ajpath.2010.100105 CrossRefPubMedPubMedCentral

37.

R.J. Kurman, M. Shih Ie, Molecular pathogenesis and extraovarian origin of epithelial ovarian cancer--shifting the paradigm. Hum Pathol 42, 918–931 (2011). doi:10.1016/j.humpath.2011.03.003 CrossRefPubMedPubMedCentral

38.

J.L. Brun, A. Feyler, G. Chene, J. Saurel, G. Brun, C. Hocke, Long-term results and prognostic factors in patients with epithelial ovarian cancer. Gynecol Oncol 78, 21–27 (2000). doi:10.1006/gyno.2000.5805 CrossRefPubMed

39.

R.G. Verhaak, P. Tamayo, J.Y. Yang, D. Hubbard, H. Zhang, C.J. Creighton, S. Fereday, M. Lawrence, S.L. Carter, C.H. Mermel, A.D. Kostic, D. Etemadmoghadam, G. Saksena, K. Cibulskis, S. Duraisamy, K. Levanon, C. Sougnez, A. Tsherniak, S. Gomez, R. Onofrio, S. Gabriel, L. Chin, N. Zhang, P.T. Spellman, Y. Zhang, R. Akbani, K.A. Hoadley, A. Kahn, M. Kobel, D. Huntsman, R.A. Soslow, A. Defazio, M.J. Birrer, J.W. Gray, J.N. Weinstein, D.D. Bowtell, R. Drapkin, J.P. Mesirov, G. Getz, D.A. Levine, M. Meyerson, Cancer genome atlas research, Prognostically relevant gene signatures of high-grade serous ovarian carcinoma. J Clin Invest 123, 517–525 (2013). doi:10.1172/JCI65833 PubMed

40.

M. Osaki, M. Oshimura, H. Ito, PI3K-Akt pathway: its functions and alterations in human cancer. Apoptosis 9, 667–676 (2004). doi:10.1023/B:APPT.0000045801.15585.dd CrossRefPubMed

41.

D.A. Altomare, H.Q. Wang, K.L. Skele, A. De Rienzo, A.J. Klein-Szanto, A.K. Godwin, J.R. Testa, AKT and mTOR phosphorylation is frequently detected in ovarian cancer and can be targeted to disrupt ovarian tumor cell growth. Oncogene 23, 5853–5857 (2004). doi:10.1038/sj.onc.1207721 CrossRefPubMed

42.

A.J. Hanrahan, N. Schultz, M.L. Westfal, R.A. Sakr, D.D. Giri, S. Scarperi, M. Janakiraman, N. Olvera, E.V. Stevens, Q.B. She, C. Aghajanian, T.A. King, E. Stanchina, D.R. Spriggs, A. Heguy, B.S. Taylor, C. Sander, N. Rosen, D.A. Levine, D.B. Solit, Genomic complexity and AKT dependence in serous ovarian cancer. Cancer Discov 2, 56–67 (2012). doi:10.1158/2159-8290.CD-11-0170 CrossRefPubMedPubMedCentral

43.

K. Kurose, X.P. Zhou, T. Araki, S.A. Cannistra, E.R. Maher, C. Eng, Frequent loss of PTEN expression is linked to elevated phosphorylated Akt levels, but not associated with p27 and cyclin D1 expression, in primary epithelial ovarian carcinomas. Am J Pathol 158, 2097–2106 (2001). doi:10.1016/S0002-9440(10)64681-0 CrossRefPubMedPubMedCentral

44.

M. Sun, G. Wang, J.E. Paciga, R.I. Feldman, Z.Q. Yuan, X.L. Ma, S.A. Shelley, R. Jove, P.N. Tsichlis, S.V. Nicosia, J.Q. Cheng, AKT1/PKBalpha kinase is frequently elevated in human cancers and its constitutive activation is required for oncogenic transformation in NIH3T3 cells. Am J Pathol 159, 431–437 (2001)CrossRefPubMedPubMedCentral

45.

Z.Q. Yuan, M. Sun, R.I. Feldman, G. Wang, X. Ma, C. Jiang, D. Coppola, S.V. Nicosia, J.Q. Cheng, Frequent activation of AKT2 and induction of apoptosis by inhibition of phosphoinositide-3-OH kinase/Akt pathway in human ovarian cancer. Oncogene 19, 2324–2330 (2000). doi:10.1038/sj.onc.1203598 CrossRefPubMed

46.

L. Shayesteh, Y. Lu, W.L. Kuo, R. Baldocchi, T. Godfrey, C. Collins, D. Pinkel, B. Powell, G.B. Mills, J.W. Gray, PIK3CA is implicated as an oncogene in ovarian cancer. Nat Genet 21, 99–102 (1999). doi:10.1038/5042 CrossRefPubMed

47.

K.E. Sheppard, C. Cullinane, K.M. Hannan, M. Wall, J. Chan, F. Barber, J. Foo, D. Cameron, A. Neilsen, P. Ng, J. Ellul, M. Kleinschmidt, K.M. Kinross, D.D. Bowtell, J.G. Christensen, R.J. Hicks, R.W. Johnstone, G.A. McArthur, R.D. Hannan, W.A. Phillips, R.B. Pearson, Synergistic inhibition of ovarian cancer cell growth by combining selective PI3K/mTOR and RAS/ERK pathway inhibitors. Eur J Cancer 49, 3936–3944 (2013). doi:10.1016/j.ejca.2013.08.007 CrossRefPubMed

48.

N. Cancer, Genome atlas research, integrated genomic analyses of ovarian carcinoma. Nature 474, 609–615 (2011). doi:10.1038/nature10166 CrossRef

49.

V. Auner, G. Kriegshauser, D. Tong, R. Horvat, A. Reinthaller, A. Mustea, R. Zeillinger, KRAS mutation analysis in ovarian samples using a high sensitivity biochip assay. BMC Cancer 9, 111 (2009). doi:10.1186/1471-2407-9-111 CrossRefPubMedPubMedCentral

50.

G. Singer, R. Oldt 3rd, Y. Cohen, B.G. Wang, D. Sidransky, R.J. Kurman, M. Shih Ie, Mutations in BRAF and KRAS characterize the development of low-grade ovarian serous carcinoma. J Natl Cancer Inst 95, 484–486 (2003)CrossRefPubMed

51.

A. Bagnato, L. Rosano, Epithelial-mesenchymal transition in ovarian cancer progression: a crucial role for the endothelin axis. Cells Tissues Organs 185, 85–94 (2007). doi:10.1159/000101307 CrossRefPubMed

52.

Y. Yang, J. Zhang, Y. Zhu, Z. Zhang, H. Sun, Y. Feng, Follicle-stimulating hormone induced epithelial-mesenchymal transition of epithelial ovarian cancer cells through follicle-stimulating hormone receptor PI3K/Akt-snail signaling pathway. Int J Gynecol Cancer 24, 1564–1574 (2014). doi:10.1097/IGC.0000000000000279 CrossRefPubMed

53.

G.Y. Zhang, A.H. Liu, G.M. Li, J.R. Wang, HPIP silencing prevents epithelial-mesenchymal transition induced by TGF-beta1 in human ovarian cancer cells. Oncol Res 24, 33–39 (2016). doi:10.3727/096504016X14575597858654 CrossRefPubMed

54.

L. Galluzzi, L. Senovilla, I. Vitale, J. Michels, I. Martins, O. Kepp, M. Castedo, G. Kroemer, Molecular mechanisms of cisplatin resistance. Oncogene 31, 1869–1883 (2012). doi:10.1038/onc.2011.384 CrossRefPubMed

55.

J. Chien, R. Kuang, C. Landen, V. Shridhar, Platinum-sensitive recurrence in ovarian cancer: the role of tumor microenvironment. Front Oncol 3, 251 (2013). doi:10.3389/fonc.2013.00251 CrossRefPubMedPubMedCentral

56.

Y. Zhang, C. Bao, Q. Mu, J. Chen, J. Wang, Y. Mi, A.J. Sayari, Y. Chen, M. Guo, Reversal of cisplatin resistance by inhibiting PI3K/Akt signal pathway in human lung cancer cells. Neoplasma 63, 362–370 (2016). doi:10.4149/304_150806N433 CrossRefPubMed

57.

C.F. Hu, Y.Y. Huang, Y.J. Wang, F.G. Gao, Upregulation of ABCG2 via the PI3K-Akt pathway contributes to acidic microenvironment-induced cisplatin resistance in A549 and LTEP-a-2 lung cancer cells. Oncol Rep 36, 455–461 (2016). doi:10.3892/or.2016.4827 CrossRefPubMed

Title: HPIP promotes epithelial-mesenchymal transition and cisplatin resistance in ovarian cancer cells through PI3K/AKT pathway activation
Authors: Suresh Bugide
Vijay Kumar Gonugunta
Vasudevarao Penugurti
Vijaya Lakshmi Malisetty
Ratna K. Vadlamudi
Bramanandam Manavathi
Publication date: 01-04-2017
Publisher: Springer Netherlands
Published in: Cellular Oncology / Issue 2/2017
Print ISSN: 2211-3428
Electronic ISSN: 2211-3436
DOI: https://doi.org/10.1007/s13402-016-0308-2

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