Inhibition of SOX4 induces melanoma cell apoptosis via downregulation of NF-κB p65 signaling

Cheng,Qiong; Du,Juan; Xie,Lin; Liu,Xiao; Li,Zheng; Zuo,Fuguo; Wu,Jinfeng; Xu,Jinhua

doi:10.3892/or.2018.6443

Inhibition of SOX4 induces melanoma cell apoptosis via downregulation of NF-κB p65 signaling

Authors:
- Qiong Cheng
- Juan Du
- Lin Xie
- Xiao Liu
- Zheng Li
- Fuguo Zuo
- Jinfeng Wu
- Jinhua Xu
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Affiliations: Department of Dermatology, East Hospital, Tongji University School of Medicine, Shanghai 200120, P.R. China, Department of Dermatology, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China
Published online on: May 16, 2018 https://doi.org/10.3892/or.2018.6443
Pages: 369-376

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Abstract

SOX4 (SRY Box 4) has attracted attention due to its important effects on cell growth, proliferation and apoptosis, among other cellular processes. However, the role of SOX4 in melanoma cell apoptosis remains unclear. In the present study, we investigated whether inhibition of SOX4 induces melanoma cell apoptosis, and explored the possible mechanism involving the NF-κB signaling pathway. SOX4 was knocked down using a lentivirus in melanoma A2058 and SK-MEL-5 cell lines. Cell proliferation was measured by MTT assay. Apoptosis was determined by flow cytometry. Western blotting was performed to detect the protein levels of SOX4, p65 and apoptosis-related proteins, such as PARP, Bcl-2, Bax and survivin. Quantitative real-time PCR (qRT-PCR) was used to examine the mRNA levels of SOX4 and p65. To determine whether SOX4 is able to bind to the promoter of p65, a CHIP-PCR assay was performed. Our data demonstrated that SOX4 knockdown significantly induced apoptosis in melanoma cells, which was accompanied by increases in cleaved PARP and Bax, and decreases in Bcl-2 and survivin. The expression of p65 was also decreased in SOX4-knockdown melanoma cells. The CHIP-PCR assay indicated that SOX4 was able to bind to the promoter region of p65. We also observed that apoptosis in SOX4-knockdown and p65-overexpressing A2058 cells was much lower than that in SOX4-knockdown alone cells. This revealed that the overexpression of p65 partially reversed SOX4 downregulation-induced apoptosis. In conclusion, our results demonstrated that inhibition of SOX4 markedly induced melanoma cell apoptosis via downregulation of the NF-κB signaling pathway, which thus may be a novel approach for the treatment of melanoma.

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1	Rhodes DR, Yu J, Shanker K, Deshpande N, Varambally R, Ghosh D, Barrette T, Pandey A and Chinnaiyan AM: Large-scale meta-analysis of cancer microarray data identifies common transcriptional profiles of neoplastic transformation and progression. Proc Natl Acad Sci USA. 101:9309–9314. 2004. View Article : Google Scholar : PubMed/NCBI
2	Zhang J, Jiang H, Shao J, Mao R, Liu J, Ma Y, Fang X, Zhao N, Zheng S and Lin B: SOX4 inhibits GBM cell growth and induces G0/G1 cell cycle arrest through Akt-p53 axis. BMC Neurol. 14:2072014. View Article : Google Scholar : PubMed/NCBI
3	Shi S, Cao X, Gu M, You B, Shan Y and You Y: Upregulated expression of SOX4 is associated with tumor growth and metastasis in nasopharyngeal carcinoma. Dis Markers. 2015:6581412015. View Article : Google Scholar : PubMed/NCBI
4	Liu P, Ramachandran S, Seyed Ali M, Scharer CD, Laycock N, Dalton WB, Williams H, Karanam S, Datta MW, Jaye DL, et al: Sex-determining region Y Box 4 is a transforming oncogene in human prostate cancer cells. Cancer Res. 66:4011–4019. 2006. View Article : Google Scholar : PubMed/NCBI
5	Yoon TM, Kim SA, Cho WS, Lee DH, Lee JK, Park YL, Lee KH, Lee JH, Kweon SS, Chung IJ, et al: SOX4 expression is associated with treatment failure and chemoradioresistance in oral squamous cell carcinoma. BMC Cancer. 15:8882015. View Article : Google Scholar : PubMed/NCBI
6	Hur W, Rhim H, Jung CK, Kim JD, Bae SH, Jang JW, Yang JM, Oh ST, Kim DG, Wang HJ, et al: SOX4 overexpression regulates the p53-mediated apoptosis in hepatocellular carcinoma: Clinical implication and functional analysis in vitro. Carcinogenesis. 31:1298–1307. 2010. View Article : Google Scholar : PubMed/NCBI
7	Pramoonjago P, Baras AS and Moskaluk CA: Knockdown of Sox4 expression by RNAi induces apoptosis in ACC3 cells. Oncogene. 25:5626–5639. 2006. View Article : Google Scholar : PubMed/NCBI
8	Bilir B, Kucuk O and Moreno CS: Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells. J Transl Med. 11:2802013. View Article : Google Scholar : PubMed/NCBI
9	Pan X, Zhao J, Zhang WN, Li HY, Mu R, Zhou T, Zhang HY, Gong WL, Yu M, Man JH, et al: Induction of SOX4 by DNA damage is critical for p53 stabilization and function. Proc Natl Acad Sci USA. 106:3788–3793. 2009. View Article : Google Scholar : PubMed/NCBI
10	Aaboe M, Birkenkamp-Demtroder K, Wiuf C, Sørensen FB, Thykjaer T, Sauter G, Jensen KM, Dyrskjøt L and Ørntoft T: SOX4 expression in bladder carcinoma: Clinical aspects and in vitro functional characterization. Cancer Res. 66:3434–3442. 2006. View Article : Google Scholar : PubMed/NCBI
11	Li J, Zhang Z and Li G: Patient outcome prediction using multiple biomarkers in human melanoma: A clinicopathological study of 118 cases. Exp Ther Med. 2:131–135. 2011. View Article : Google Scholar : PubMed/NCBI
12	Jafarnejad SM, Wani AA, Martinka M and Li G: Prognostic significance of Sox4 expression in human cutaneous melanoma and its role in cell migration and invasion. Am J Pathol. 177:2741–2752. 2010. View Article : Google Scholar : PubMed/NCBI
13	Watanabe M, Umezawa K, Higashihara M and Horie R: Combined inhibition of NF-κB and Bcl-2 triggers synergistic reduction of viability and induces apoptosis in melanoma cells. Oncol Res. 21:173–180. 2013. View Article : Google Scholar : PubMed/NCBI
14	Vervoort SJ, van Boxtel R and Coffer PJ: The role of SRY-related HMG box transcription factor 4 (SOX4) in tumorigenesis and metastasis: Friend or foe? Oncogene. 32:3397–3409. 2013. View Article : Google Scholar : PubMed/NCBI
15	Ahn SG, Kim HS, Jeong SW, Kim BE, Rhim H, Shim JY, Kim JW, Lee JH and Kim IK: Sox-4 is a positive regulator of Hep3B and HepG2 cells' apoptosis induced by prostaglandin (PG)A₂ and Δ¹²-PGJ₂. Exp Mol Med. 34:243–249. 2002. View Article : Google Scholar : PubMed/NCBI
16	Ahn SG, Cho GH, Jeong SY, Rhim H, Choi JY and Kim IK: Identification of cDNAs for Sox-4, an HMG-Box protein, and a novel human homolog of yeast splicing factor SSF-1 differentially regulated during apoptosis induced by prostaglandin A₂/Δ¹²-PGJ₂ in Hep3B cells. Biochem Biophys Res Commun. 260:216–221. 1999. View Article : Google Scholar : PubMed/NCBI
17	Zhou Y, Wang X, Huang Y, Chen Y, Zhao G, Yao Q, Jin C, Huang Y, Liu X and Li G: Down-regulated SOX4 expression suppresses cell proliferation, metastasis and induces apoptosis in Xuanwei female lung cancer patients. J Cell Biochem. 116:1007–1018. 2015. View Article : Google Scholar : PubMed/NCBI
18	Jang SM, Kang EJ, Kim JW, Kim CH, An JH and Choi KH: Transcription factor Sox4 is required for PUMA-mediated apoptosis induced by histone deacetylase inhibitor, TSA. Biochem Biophys Res Commun. 438:445–451. 2013. View Article : Google Scholar : PubMed/NCBI
19	Weng HL and Wang MJ: Effects of microRNA-338-3p on morphine-induced apoptosis and its underlying mechanisms. Mol Med Rep. 14:2085–2092. 2016. View Article : Google Scholar : PubMed/NCBI
20	Hur EH, Hur W, Choi JY, Kim IK, Kim HY, Yoon SK and Rhim H: Functional identification of the pro-apoptotic effector domain in human Sox4. Biochem Biophys Res Commun. 325:59–67. 2004. View Article : Google Scholar : PubMed/NCBI
21	Murtas D, Piras F, Minerba L, Ugalde J, Piga M, Maxia C, Perra MT and Sirigu P: Nuclear factor-κB expression is predictive of overall survival in patients with cutaneous melanoma. Oncol Lett. 1:633–639. 2010. View Article : Google Scholar : PubMed/NCBI
22	Karin M, Cao Y, Greten FR and Li ZW: NF-kappaB in cancer: From innocent bystander to major culprit. Nat Rev Cancer. 2:301–310. 2002. View Article : Google Scholar : PubMed/NCBI
23	Park MH and Hong JT: Roles of NF-κB in cancer and inflammatory diseases and their therapeutic approaches. Cells. 5:152016. View Article : Google Scholar :
24	Savva CG, Totokotsopoulos S, Nicolaou KC, Neophytou CM and Constantinou AI: Selective activation of TNFR1 and NF-κB inhibition by a novel biyouyanagin analogue promotes apoptosis in acute leukemia cells. BMC Cancer. 16:2792016. View Article : Google Scholar : PubMed/NCBI
25	Pahl HL: Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene. 18:6853–6866. 1999. View Article : Google Scholar : PubMed/NCBI
26	Tao Y, Zhang ML, Ma PC, Sun JF, Zhou WQ, Cao YP and Li LJ: Triptolide inhibits proliferation and induces apoptosis of human melanoma A375 cells. Asian Pac J Cancer Prev. 13:1611–1615. 2012. View Article : Google Scholar : PubMed/NCBI
27	Schneider A, Martin-Villalba A, Weih F, Vogel J, Wirth T and Schwaninger M: NF-kappaB is activated and promotes cell death in focal cerebral ischemia. Nat Med. 5:554–559. 1999. View Article : Google Scholar : PubMed/NCBI
28	Li F and Ling X: Survivin study: An update of ‘what is the next wave’? J Cell Physiol. 208:476–486. 2006. View Article : Google Scholar : PubMed/NCBI
29	Grossman D, McNiff JM, Li F and Altieri DC: Expression and targeting of the apoptosis inhibitor, survivin, in human melanoma. J Invest Dermatol. 113:1076–1081. 1999. View Article : Google Scholar : PubMed/NCBI
30	Marusawa H, Matsuzawa S, Welsh K, Zou H, Armstrong R, Tamm I and Reed JC: HBXIP functions as a cofactor of survivin in apoptosis suppression. EMBO J. 22:2729–2740. 2003. View Article : Google Scholar : PubMed/NCBI