p12CDK2-AP1 interacts with CD82 to regulate the proliferation and survival of human oral squamous cell carcinoma cells

Chai,Juan; Ju,Jun; Zhang,Shao-Wu; Shen,Zhi-Yuan; Liang,Liang; Yang,Xiang-Ming; Ma,Chao; Ni,Qian-Wei; Sun,Mo-Yi

doi:10.3892/or.2016.4893

p12CDK2-AP1 interacts with CD82 to regulate the proliferation and survival of human oral squamous cell carcinoma cells

Authors:
- Juan Chai
- Jun Ju
- Shao-Wu Zhang
- Zhi-Yuan Shen
- Liang Liang
- Xiang-Ming Yang
- Chao Ma
- Qian-Wei Ni
- Mo-Yi Sun
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Affiliations: State Key Laboratory of Military Stomatology, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, College of Automation, Northwestern Polytechnical University, Xi'an, Shaanxi 710072, P.R. China
Published online on: June 22, 2016 https://doi.org/10.3892/or.2016.4893
Pages: 737-744

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Abstract

p12 cyclin-dependent kinase 2 (CDK2)-associating protein 1 (p12CDK2-AP1) has been demonstrated to negatively regulate the activity of CDK2. However, the underlying molecular mechanism remains largely unknown. We aimed to determine the potential binding proteins of p12CDK2-AP1 and to elucidate the role of p12CDK2-AP1 in the regulation of the proliferation, invasion, apoptosis, and in vivo growth of human oral squamous cell carcinoma cells. The protein-protein interaction was predicted using computational decision templates. The predicted p12CDK2‑AP1 interacting proteins were overexpressed in human oral squamous cell carcinoma OSCC-15 cells, and the protein binding was examined using co-precipitation (Co-IP). Cell proliferation and invasion were determined via MTT assay and Transwell system, respectively. Cell apoptosis was evaluated using Annexin V-FITC/PI double staining followed by flow cytometric analysis. The in vivo growth of OSCC-15 cells was examined in nude mouse tumor xenografts. We found that overexpression of either p12CDK2-AP1 or CD82 significantly suppressed the proliferation and invasion but promoted the apoptosis of OSCC-15 cells (P<0.05). Importantly, combined overexpression of p12CDK2-AP1 and CD82 showed synergistic antitumor activity compared with the overexpression of a single protein alone (P<0.05). Additionally, the simultaneous overexpression of p12CDK2-AP1 and CD82 significantly suppressed the in vivo tumor growth of OSCC-15 cells in nude mice compared with the negative control (P<0.05). Our findings indicate that p12CDK2-AP1 interacts with CD82 to play a functional role in suppressing the in vitro and in vivo growth of OSCC-15 cells.

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1	Marcu LG and Yeoh E: A review of risk factors and genetic alterations in head and neck carcinogenesis and implications for current and future approaches to treatment. J Cancer Res Clin Oncol. 135:1303–1314. 2009. View Article : Google Scholar : PubMed/NCBI
2	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar : PubMed/NCBI
3	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
4	Chen C, Méndez E, Houck J, Fan W, Lohavanichbutr P, Doody D, Yueh B, Futran ND, Upton M, Farwell DG, et al: Gene expression profiling identifies genes predictive of oral squamous cell carcinoma. Cancer Epidemiol Biomarkers Prev. 17:2152–2162. 2008. View Article : Google Scholar : PubMed/NCBI
5	Todd R, McBride J, Tsuji T, Donoff RB, Nagai M, Chou MY, Chiang T and Wong DT: Deleted in oral cancer-1 (doc-1), a novel oral tumor suppressor gene. FASEB J. 9:1362–1370. 1995.PubMed/NCBI
6	Tsuji T, Duh FM, Latif F, Popescu NC, Zimonjic DB, McBride J, Matsuo K, Ohyama H, Todd R, Nagata E, et al: Cloning, mapping, expression, function, and mutation analyses of the human ortholog of the hamster putative tumor suppressor gene Doc-1. J Biol Chem. 273:6704–6709. 1998. View Article : Google Scholar : PubMed/NCBI
7	Shintani S, Mihara M, Terakado N, Nakahara Y, Matsumura T, Kohno Y, Ohyama H, McBride J, Kent R, Todd R, et al: Reduction of p12^DOC-1 expression is a negative prognostic indicator in patients with surgically resected oral squamous cell carcinoma. Clin Cancer Res. 7:2776–2782. 2001.PubMed/NCBI
8	Kohno Y, Patel V, Kim Y, Tsuji T, Chin BR, Sun M, Bruce Donoff R, Kent R, Wong D and Todd R: Apoptosis, proliferation and p12(doc-1) profiles in normal, dysplastic and malignant squamous epithelium of the Syrian hamster cheek pouch model. Oral Oncol. 38:274–280. 2002. View Article : Google Scholar : PubMed/NCBI
9	Matsuo K, Shintani S, Tsuji T, Nagata E, Lerman M, McBride J, Nakahara Y, Ohyama H, Todd R and Wong DT: p12(DOC-1), a growth suppressor, associates with DNA polymerase alpha/primase. FASEB J. 14:1318–1324. 2000. View Article : Google Scholar : PubMed/NCBI
10	Buajeeb W, Zhang X, Ohyama H, Han D, Surarit R, Kim Y and Wong DT: Interaction of the CDK2-associated protein-1, p12(DOC-1/CDK2AP1), with its homolog, p14(DOC-1R). Biochem Biophys Res Commun. 315:998–1003. 2004. View Article : Google Scholar : PubMed/NCBI
11	Zhang SW, Hao LY and Zhang TH: Prediction of protein-protein interaction with pairwise kernel support vector machine. Int J Mol Sci. 15:3220–3233. 2014. View Article : Google Scholar : PubMed/NCBI
12	Hue M, Riffle M, Vert JP and Noble WS: Large-scale prediction of protein-protein interactions from structures. BMC Bioinformatics. 11:1442010. View Article : Google Scholar : PubMed/NCBI
13	Craig RA and Liao L: Improving protein protein interaction prediction based on phylogenetic information using a least-squares support vector machine. Ann NY Acad Sci. 1115:154–167. 2007. View Article : Google Scholar : PubMed/NCBI
14	Chen W, Zhang SW, Cheng YM and Pan Q: Prediction of protein-protein interaction types using the decision templates based on multiple classier fusion. Math Comput Model. 52:2075–2084. 2010. View Article : Google Scholar
15	Todd R, Hinds PW, Munger K, Rustgi AK, Opitz OG, Suliman Y and Wong DT: Cell cycle dysregulation in oral cancer. Crit Rev Oral Biol Med. 13:51–61. 2002. View Article : Google Scholar : PubMed/NCBI
16	Shintani S, Ohyama H, Zhang X, McBride J, Matsuo K, Tsuji T, Hu MG, Hu G, Kohno Y, Lerman M, et al: p12(DOC-1) is a novel cyclin-dependent kinase 2-associated protein. Mol Cell Biol. 20:6300–6307. 2000. View Article : Google Scholar : PubMed/NCBI
17	Zhou W, Guan X, Wang L, Liao Y and Huang J: p12(CDK2-AP1) inhibits breast cancer cell proliferation and in vivo tumor growth. J Cancer Res Clin Oncol. 138:2085–2093. 2012. View Article : Google Scholar : PubMed/NCBI
18	Choi MG, Sohn TS, Park SB, Paik YH, Noh JH, Kim KM, Park CK and Kim S: Decreased expression of p12 is associated with more advanced tumor invasion in human gastric cancer tissues. Eur Surg Res. 42:223–229. 2009. View Article : Google Scholar : PubMed/NCBI
19	Hiyoshi Y, Watanabe M, Hirashima K, Karashima R, Sato N, Imamura Y, Nagai Y, Yoshida N, Toyama E, Hayashi N, et al: p12^CDK2-AP1 is associated with tumor progression and a poor prognosis in esophageal squamous cell carcinoma. Oncol Rep. 22:35–39. 2009.PubMed/NCBI
20	Sun M, Zheng J, Xue H, Jiang Y, Li C, Li J, Jin W, Shen M, Yang X and Ni Q: Silencing P12^CDK2AP1 with a lentivirus promotes HaCaT cell proliferation. Mol Med Rep. 7:471–475. 2013.
21	Zheng J, Xue H, Wang T, Jiang Y, Liu B, Li J, Liu Y, Wang W, Zhang B and Sun M: miR-21 downregulates the tumor suppressor P12 CDK2AP1 and stimulates cell proliferation and invasion. J Cell Biochem. 112:872–880. 2011. View Article : Google Scholar : PubMed/NCBI
22	Liu L, Yang X, Ni Q, Xiao Z, Zhao Y, Han J, Sun M and Chen B: Interaction between p12^CDK2AP1 and a novel unnamed protein product inhibits cell proliferation by regulating the cell cycle. Mol Med Rep. 9:156–162. 2014.
23	Dong JT, Lamb PW, Rinker-Schaeffer CW, Vukanovic J, Ichikawa T, Isaacs JT and Barrett JC: KAI1, a metastasis suppressor gene for prostate cancer on human chromosome 11p11.2. Science. 268:884–886. 1995. View Article : Google Scholar : PubMed/NCBI
24	Tang Y, Cheng Y, Martinka M, Ong CJ and Li G: Prognostic significance of KAI1/CD82 in human melanoma and its role in cell migration and invasion through the regulation of ING4. Carcinogenesis. 35:86–95. 2014. View Article : Google Scholar
25	Rowe A and Jackson P: Expression of KITENIN, a KAI1/CD82 binding protein and metastasis enhancer, in bladder cancer cell lines: Relationship to KAI1/CD82 levels and invasive behaviour. Oncol Rep. 16:1267–1272. 2006.PubMed/NCBI
26	Malik FA, Sanders AJ, Kayani MA and Jiang WG: Effect of expressional alteration of KAI1 on breast cancer cell growth, adhesion, migration and invasion. Cancer Genomics Proteomics. 6:205–213. 2009.PubMed/NCBI
27	Pal'tseva EM, Samofalova OIu, Gorbacheva IuV and Tsar'kov PV: Expression of some biomarkers in primary colon adenocarcinomas and their lymph node metastases. Arkh Patol. 74:12–18. 2012.In Russian. PubMed/NCBI
28	Friess H, Guo XZ, Tempia-Caliera AA, Fukuda A, Martignoni ME, Zimmermann A, Korc M and Büchler MW: Differential expression of metastasis-associated genes in papilla of vater and pancreatic cancer correlates with disease stage. J Clin Oncol. 19:2422–2432. 2001.PubMed/NCBI
29	Choi UJ, Jee BK, Lim Y and Lee KH: KAI1/CD82 decreases Rac1 expression and cell proliferation through PI3K/Akt/mTOR pathway in H1299 lung carcinoma cells. Cell Biochem Funct. 27:40–47. 2009. View Article : Google Scholar
30	Shiwu WU, Lan Y, Wenqing S, Lei Z and Yisheng T: Expression and clinical significance of CD82/KAI1 and E-cadherin in non-small cell lung cancer. Arch Iran Med. 15:707–712. 2012.PubMed/NCBI
31	Liu FS, Dong JT, Chen JT, Hsieh YT, Ho ES, Hung MJ, Lu CH and Chiou LC: KAI1 metastasis suppressor protein is down-regulated during the progression of human endometrial cancer. Clin Cancer Res. 9:1393–1398. 2003.PubMed/NCBI
32	Miyazaki T, Kato H, Shitara Y, Yoshikawa M, Tajima K, Masuda N, Shouji H, Tsukada K, Nakajima T and Kuwano H: Mutation and expression of the metastasis suppressor gene KAI1 in esophageal squamous cell carcinoma. Cancer. 89:955–962. 2000. View Article : Google Scholar : PubMed/NCBI
33	Imai Y, Sasaki T, Shinagawa Y, Akimoto K and Fujibayashi T: Expression of metastasis suppressor gene (KAI1/CD82) in oral squamous cell carcinoma and its clinico-pathological significance. Oral Oncol. 38:557–561. 2002. View Article : Google Scholar : PubMed/NCBI
34	Farhadieh RD, Smee R, Ow K, Yang JL, Russell PJ, Crouch R, Jackson P and Jacobson IV: Down-regulation of KAI1/CD82 protein expression in oral cancer correlates with reduced disease free survival and overall patient survival. Cancer Lett. 213:91–98. 2004. View Article : Google Scholar : PubMed/NCBI
35	Kawasaki G, Yoshitomi I, Yanamoto S, Yamada S, Mizuno A and Umeda M: Expression of thymidylate synthase and dihydropyrimidine dehydrogenase in primary oral squamous cell carcinoma and corresponding metastases in cervical lymph nodes: Association with the metastasis suppressor CD82. Anticancer Res. 31:3521–3526. 2011.PubMed/NCBI
36	Abe M, Sugiura T, Takahashi M, Ishii K, Shimoda M and Shirasuna K: A novel function of CD82/KAI-1 on E-cadherin-mediated homophilic cellular adhesion of cancer cells. Cancer Lett. 266:163–170. 2008. View Article : Google Scholar : PubMed/NCBI
37	Takahashi M, Sugiura T, Abe M, Ishii K and Shirasuna K: Regulation of c-Met signaling by the tetraspanin KAI-1/CD82 affects cancer cell migration. Int J Cancer. 121:1919–1929. 2007. View Article : Google Scholar : PubMed/NCBI