Valproic acid exhibits different cell growth arrest effect in three HPV-positive/negative cervical cancer cells and possibly via inducing Notch1 cleavage and E6 downregulation

Feng,Shuyu; Yang,Yue; Lv,Jingyi; Sun,Lichun; Liu,Mingqiu

doi:10.3892/ijo.2016.3508

Valproic acid exhibits different cell growth arrest effect in three HPV-positive/negative cervical cancer cells and possibly via inducing Notch1 cleavage and E6 downregulation

Authors:
- Shuyu Feng
- Yue Yang
- Jingyi Lv
- Lichun Sun
- Mingqiu Liu
View Affiliations

Affiliations: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, P.R. China, Department of Medicine, School of Medicine, Tulane Health Sciences Center, New Orleans, LA 70112-2699, USA
Published online on: May 6, 2016 https://doi.org/10.3892/ijo.2016.3508
Pages: 422-430

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

We investigated the effect of valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, and the mechanism of VPA-induced growth inhibition on three cervical cancer cell lines with different molecular and genetic background. We found that VPA induced proliferation suppression, cell apoptosis and cell cycle arrest in all tested cell lines, with an increase of Notch1 active form ICN1 as a tumor suppressor and its target gene HES1. Noteworthy, blocking of Notch signaling with DAPT resulted in growth inhibition in ICN1-overexpressing CaSki and HT-3 cells. Thus, endogenous Notch signaling may be necessary for survival of ICN1-overexpressing cervical cancer cell lines. Furthermore, G1 phase arrest was induced in HeLa and CaSki cells by VPA while G2 phase arrest was induced in HT-3 cells, suggesting different mechanism in this cycle arrest. We also found VPA suppressed oncogene E6 in a Notch-independent manner, and induced significant apoptosis in E6-overexpressing HPV positive CaSki cells. Cell morphological change was also observed in HeLa and HT-3 cell lines after VPA treatment with an upregulation of EMT transcription factor Snail1. Notch signaling inhibitor DAPT partly reversed VPA-induced Snail1 upregulation in HeLa cells. This discovery supports that VPA may induce EMT at least partly via Notch activation.

View Figures

View References

1	Walboomers JM, Jacobs MV, Manos MM, Bosch FX, Kummer JA, Shah KV, Snijders PJ, Peto J, Meijer CJ and Muñoz N: Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol. 189:12–19. 1999. View Article : Google Scholar : PubMed/NCBI
2	Wu L and Griffin JD: Modulation of Notch signaling by mastermind-like (MAML) transcriptional co-activators and their involvement in tumorigenesis. Semin Cancer Biol. 14:348–356. 2004. View Article : Google Scholar : PubMed/NCBI
3	Weijzen S, Zlobin A, Braid M, Miele L and Kast WM: HPV16 E6 and E7 oncoproteins regulate Notch-1 expression and cooperate to induce transformation. J Cell Physiol. 194:356–362. 2003. View Article : Google Scholar : PubMed/NCBI
4	Schwarz JK, Payton JE, Rashmi R, Xiang T, Jia Y, Huettner P, Rogers BE, Yang Q, Watson M, Rader JS, et al: Pathway-specific analysis of gene expression data identifies the PI3K/Akt pathway as a novel therapeutic target in cervical cancer. Clin Cancer Res. 18:1464–1471. 2012. View Article : Google Scholar : PubMed/NCBI
5	Perez-Plasencia C, Duenas-Gonzalez A and Alatorre-Tavera B: Second hit in cervical carcinogenesis process: Involvement of wnt/beta catenin pathway. Int Arch Med. 1:102008. View Article : Google Scholar : PubMed/NCBI
6	Yousif NG, Sadiq AM, Yousif MG, Al-Mudhafar RH, Al-Baghdadi JJ and Hadi N: Notch1 ligand signaling pathway activated in cervical cancer: Poor prognosis with high-level JAG1/Notch1. Arch Gynecol Obstet. 292:899–904. 2015. View Article : Google Scholar : PubMed/NCBI
7	Maliekal TT, Bajaj J, Giri V, Subramanyam D and Krishna S: The role of Notch signaling in human cervical cancer: Implications for solid tumors. Oncogene. 27:5110–5114. 2008. View Article : Google Scholar : PubMed/NCBI
8	Wu L, Aster JC, Blacklow SC, Lake R, Artavanis-Tsakonas S and Griffin JD: MAML1, a human homologue of Drosophila mastermind, is a transcriptional co-activator for NOTCH receptors. Nat Genet. 26:484–489. 2000. View Article : Google Scholar : PubMed/NCBI
9	Wael H, Yoshida R, Kudoh S, Hasegawa K, Niimori-Kita K and Ito T: Notch1 signaling controls cell proliferation, apoptosis and differentiation in lung carcinoma. Lung Cancer. 85:131–140. 2014. View Article : Google Scholar : PubMed/NCBI
10	Brakenhoff RH: Cancer. Another NOTCH for cancer. Science. 333:1102–1103. 2011. View Article : Google Scholar : PubMed/NCBI
11	del Campo JM, Prat A, Gil-Moreno A, Pérez J and Parera M: Update on novel therapeutic agents for cervical cancer. Gynecol Oncol. 110(Suppl 2): S72–S76. 2008. View Article : Google Scholar : PubMed/NCBI
12	Pant S, Jones SF, Kurkjian CD, Infante JR, Moore KN, Burris HA, McMeekin DS, Benhadji KA, Patel BK, Frenzel MJ, et al: A first-in-human phase I study of the oral Notch inhibitor, LY900009, in patients with advanced cancer. Eur J Cancer. 56:1–9. 2016. View Article : Google Scholar : PubMed/NCBI
13	Kong R, Feng J, Ma Y, Zhou B, Li S, Zhang W, Jiang J, Zhang J, Qiao Z, Zhang T, et al: Silencing NACK by siRNA inhibits tumorigenesis in non-small cell lung cancer via targeting Notch1 signaling pathway. Oncol Rep. 35:2306–2314. 2016.PubMed/NCBI
14	Duenas-Gonzalez A, Candelaria M, Perez-Plascencia C, Perez-Cardenas E, de la Cruz-Hernandez E and Herrera LA: Valproic acid as epigenetic cancer drug: Preclinical, clinical and transcriptional effects on solid tumors. Cancer Treat Rev. 34:206–222. 2008. View Article : Google Scholar : PubMed/NCBI
15	Ranganathan P, Weaver KL and Capobianco AJ: Notch signalling in solid tumours: A little bit of everything but not all the time. Nat Rev Cancer. 11:338–351. 2011. View Article : Google Scholar : PubMed/NCBI
16	Zagouras P, Stifani S, Blaumueller CM, Carcangiu ML and Artavanis-Tsakonas S: Alterations in Notch signaling in neoplastic lesions of the human cervix. Proc Natl Acad Sci USA. 92:6414–6418. 1995. View Article : Google Scholar : PubMed/NCBI
17	Tewari KS, Taylor JA, Liao SY, DiSaia PJ, Burger RA, Monk BJ, Hughes CC and Villarreal LP: Development and assessment of a general theory of cervical carcinogenesis utilizing a severe combined immunodeficiency murine-human xenograft model. Gynecol Oncol. 77:137–148. 2000. View Article : Google Scholar : PubMed/NCBI
18	Talora C, Sgroi DC, Crum CP and Dotto GP: Specific down-modulation of Notch1 signaling in cervical cancer cells is required for sustained HPV-E6/E7 expression and late steps of malignant transformation. Genes Dev. 16:2252–2263. 2002. View Article : Google Scholar : PubMed/NCBI
19	Talora C, Cialfi S, Segatto O, Morrone S, Kim Choi J, Frati L, Paolo Dotto G, Gulino A and Screpanti I: Constitutively active Notch1 induces growth arrest of HPV-positive cervical cancer cells via separate signaling pathways. Exp Cell Res. 305:343–354. 2005. View Article : Google Scholar : PubMed/NCBI
20	Yu H, Zhao X, Huang S, Jian L, Qian G and Ge S: Blocking Notch1 signaling by RNA interference can induce growth inhibition in HeLa cells. Int J Gynecol Cancer. 17:511–516. 2007. View Article : Google Scholar : PubMed/NCBI
21	Khan O and La Thangue NB: HDAC inhibitors in cancer biology: Emerging mechanisms and clinical applications. Immunol Cell Biol. 90:85–94. 2012. View Article : Google Scholar
22	Huang BH, Laban M, Leung CH, Lee L, Lee CK, Salto-Tellez M, Raju GC and Hooi SC: Inhibition of histone deacetylase 2 increases apoptosis and p21Cip1/WAF1 expression, independent of histone deacetylase 1. Cell Death Differ. 12:395–404. 2005. View Article : Google Scholar : PubMed/NCBI
23	Teng HF, Li PN, Hou DR, Liu SW, Lin CT, Loo MR, Kao CH, Lin KH and Chen SL: Valproic acid enhances Oct4 promoter activity through PI3K/Akt/mTOR pathway activated nuclear receptors. Mol Cell Endocrinol. 383:147–158. 2014. View Article : Google Scholar
24	Shan Z, Feng-Nian R, Jie G and Ting Z: Effects of valproic acid on proliferation, apoptosis, angiogenesis and metastasis of ovarian cancer in vitro and in vivo. Asian Pac J Cancer Prev. 13:3977–3982. 2012. View Article : Google Scholar : PubMed/NCBI
25	Dotto GP: Notch tumor suppressor function. Oncogene. 27:5115–5123. 2008. View Article : Google Scholar : PubMed/NCBI
26	Franko-Tobin LG, Mackey LV, Huang W, Song X, Jin B, Luo J, Morris LM, Liu M, Fuselier JA, Coy DH, et al: Notch1-mediated tumor suppression in cervical cancer with the involvement of SST signaling and its application in enhanced SSTR-targeted therapeutics. Oncologist. 17:220–232. 2012. View Article : Google Scholar : PubMed/NCBI
27	Tsai C, Leslie JS, Franko-Tobin LG, Prasnal MC, Yang T, Vienna Mackey L, Fuselier JA, Coy DH, Liu M, Yu C, et al: Valproic acid suppresses cervical cancer tumor progression possibly via activating Notch1 signaling and enhances receptor-targeted cancer chemotherapeutic via activating somatostatin receptor type II. Arch Gynecol Obstet. 288:393–400. 2013. View Article : Google Scholar : PubMed/NCBI
28	Greenblatt DY, Cayo MA, Adler JT, Ning L, Haymart MR, Kunnimalaiyaan M and Chen H: Valproic acid activates Notch1 signaling and induces apoptosis in medullary thyroid cancer cells. Ann Surg. 247:1036–1040. 2008. View Article : Google Scholar : PubMed/NCBI
29	Greenblatt DY, Vaccaro AM, Jaskula-Sztul R, Ning L, Haymart M, Kunnimalaiyaan M and Chen H: Valproic acid activates notch-1 signaling and regulates the neuroendocrine phenotype in carcinoid cancer cells. Oncologist. 12:942–951. 2007. View Article : Google Scholar : PubMed/NCBI
30	Platta CS, Greenblatt DY, Kunnimalaiyaan M and Chen H: Valproic acid induces Notch1 signaling in small cell lung cancer cells. J Surg Res. 148:31–37. 2008. View Article : Google Scholar : PubMed/NCBI
31	Kuncharin Y, Sangphech N, Kueanjinda P, Bhattarakosol P and Palaga T: MAML1 regulates cell viability via the NF-κB pathway in cervical cancer cell lines. Exp Cell Res. 317:1830–1840. 2011. View Article : Google Scholar : PubMed/NCBI
32	Lamouille S, Xu J and Derynck R: Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 15:178–196. 2014. View Article : Google Scholar : PubMed/NCBI
33	Gonzalez DM and Medici D: Signaling mechanisms of the epithelial-mesenchymal transition. Sci Signal. 7:re82014. View Article : Google Scholar : PubMed/NCBI
34	Feng J, Cen J, Li J, Zhao R, Zhu C, Wang Z, Xie J and Tang W: Histone deacetylase inhibitor valproic acid (VPA) promotes the epithelial mesenchymal transition of colorectal cancer cells via up regulation of Snail. Cell Adhes Migr. 9:495–501. 2015. View Article : Google Scholar
35	Mawatari T, Ninomiya I, Inokuchi M, Harada S, Hayashi H, Oyama K, Makino I, Nakagawara H, Miyashita T, Tajima H, et al: Valproic acid inhibits proliferation of HER2-expressing breast cancer cells by inducing cell cycle arrest and apoptosis through Hsp70 acetylation. Int J Oncol. 47:2073–2081. 2015.PubMed/NCBI
36	Kostrouchová M, Kostrouch Z and Kostrouchová M: Valproic acid, a molecular lead to multiple regulatory pathways. Folia Biol (Praha). 53:37–49. 2007.
37	Qing H, He G, Ly PT, Fox CJ, Staufenbiel M, Cai F, Zhang Z, Wei S, Sun X, Chen CH, et al: Valproic acid inhibits Abeta production, neuritic plaque formation, and behavioral deficits in Alzheimer's disease mouse models. J Exp Med. 205:2781–2789. 2008. View Article : Google Scholar : PubMed/NCBI
38	Dueñas-González A, Lizano M, Candelaria M, Cetina L, Arce C and Cervera E: Epigenetics of cervical cancer. An overview and therapeutic perspectives. Mol Cancer. 4:382005. View Article : Google Scholar : PubMed/NCBI
39	Wang L, Qin H, Chen B, Xin X, Li J and Han H: Overexpressed active Notch1 induces cell growth arrest of HeLa cervical carcinoma cells. Int J Gynecol Cancer. 17:1283–1292. 2007. View Article : Google Scholar : PubMed/NCBI
40	Yuan CH, Filippova M, Krstenansky JL and Duerksen-Hughes PJ: Flavonol and imidazole derivatives block HPV16 E6 activities and reactivate apoptotic pathways in HPV(+) cells. Cell Death Dis. 7:20602016. View Article : Google Scholar
41	Cai Q, Lv L, Shao Q, Li X and Dian A: Human papillomavirus early proteins and apoptosis. Arch Gynecol Obstet. 287:541–548. 2013. View Article : Google Scholar