Abstract
Deregulated cell cycle can contribute to the unscheduled proliferation in cancer cells. Overexpression of cell cycle regulators CDK4 and Cyclin D1 has been reported in many cancers. The aim of this study is to determine the clinical implications of CDK4 and Cyclin D1 in hepatocellular carcinoma (HCC). The levels of mRNA and protein were analyzed by quantitative real-time RT-PCR and immunohistochemistry, respectively, in 59 paired HCC and the neighboring noncancer tissues. The relationship between CDK4 and Cyclin D1 expression, clinicopathological parameters, and prognosis was investigated. Our data demonstrated that the mRNA level of CDK4 was up-regulated (p = 0.019), while that of Cyclin D1 was down-regulated (p = 0.002), in HCC. Immunohistochemical data confirmed that CDK4 protein was increased in 73 % and Cyclin D1 protein was decreased in 66 % of HCC samples. Overexpression of CDK4 was correlated with HBV (p = 0.054, borderline significant), tumor size (p = 0.014), and stage (p = 0.010). The Kaplan–Meier survival curves showed that high CDK4 was correlated with a poor survival rate (I vs. II, p < 0.001; I vs. III, p < 0.001). Univariate analysis showed that tumor size (p = 0.002), stage (p = 0.021), and high CDK4 score (I vs. II–III, p < 0.001) were significant prognostic factors. Multivariate analysis showed that tumor size (p = 0.007) and high CDK4 score (I vs. II–III, p < 0.001) were independent factors for overall survival of HCC. The expression of Cyclin D1 was not correlated with CDK4 expression, tumor grades, survival rate, and any clinicopathological parameters. CDK4 could provide a clinical prognostic marker for HCC progression.
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References
Pardee AB. G1 events and regulation of cell proliferation. Science. 1989;246(4930):603–8.
Bartek J, Bartkova J, Lukas J. The retinoblastoma protein pathway and the restriction point. Curr Opin Cell Biol. 1996;8(6):805–14.
Malumbres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009;9(3):153–66.
Malumbres M, Barbacid M. To cycle or not to cycle: a critical decision in cancer. Nat Rev Cancer. 2001;1(3):222–31.
Malumbres M, Sotillo R, Santamaria D, Galan J, Cerezo A, Ortega S, et al. Mammalian cells cycle without the D-type cyclin-dependent kinases Cdk4 and Cdk6. Cell. 2004;118(4):493–504.
Ortega S, Prieto I, Odajima J, Martin A, Dubus P, Sotillo R, et al. Cyclin-dependent kinase 2 is essential for meiosis but not for mitotic cell division in mice. Nat Genet. 2003;35(1):25–31.
Baker SJ. A role for Cdk4 in angiogenesis. Cell Cycle. 2010;9(13):2493.
Diehl JA. Cycling to cancer with cyclin D1. Cancer Biol Ther. 2002;1(3):226–31.
Wu A, Wu B, Guo J, Luo W, Wu D, Yang H, et al. Elevated expression of CDK4 in lung cancer. J Transl Med. 2011;9(38):38.
Dobashi Y, Goto A, Fukayama M, Abe A, Ooi A. Overexpression of cdk4/cyclin D1, a possible mediator of apoptosis and an indicator of prognosis in human primary lung carcinoma. Int J Cancer. 2004;110(4):532–41.
Poomsawat S, Buajeeb W, Khovidhunkit SO, Punyasingh J. Alteration in the expression of cdk4 and cdk6 proteins in oral cancer and premalignant lesions. J Oral Pathol Med. 2010;39(10):793–9.
Yu Q, Sicinska E, Geng Y, Ahnstrom M, Zagozdzon A, Kong Y, et al. Requirement for CDK4 kinase function in breast cancer. Cancer Cell. 2006;9(1):23–32.
An HX, Beckmann MW, Reifenberger G, Bender HG, Niederacher D. Gene amplification and overexpression of CDK4 in sporadic breast carcinomas is associated with high tumor cell proliferation. Am J Pathol. 1999;154(1):113–8.
He J, Olson JJ, James CD. Lack of p16INK4 or retinoblastoma protein (pRb), or amplification-associated overexpression of cdk4 is observed in distinct subsets of malignant glial tumors and cell lines. Cancer Res. 1995;55(21):4833–6.
Lam PY, Di Tomaso E, Ng HK, Pang JC, Roussel MF, Hjelm NM. Expression of p19INK4d, CDK4, CDK6 in glioblastoma multiforme. Br J Neurosurg. 2000;14(1):28–32.
Dei Tos AP, Doglioni C, Piccinin S, Sciot R, Furlanetto A, Boiocchi M, et al. Coordinated expression and amplification of the MDM2, CDK4, and HMGI-C genes in atypical lipomatous tumours. J Pathol. 2000;190(5):531–6.
Masaki T, Shiratori Y, Rengifo W, Igarashi K, Yamagata M, Kurokohchi K, et al. Cyclins and cyclin-dependent kinases: comparative study of hepatocellular carcinoma versus cirrhosis. Hepatology (Baltimore, Md.) 2003;37(3):534–43.
Lindberg D, Hessman O, Akerstrom G, Westin G. Cyclin-dependent kinase 4 (CDK4) expression in pancreatic endocrine tumors. Neuroendocrinology. 2007;86(2):112–8.
Kanoe H, Nakayama T, Murakami H, Hosaka T, Yamamoto H, Nakashima Y, et al. Amplification of the CDK4 gene in sarcomas: tumor specificity and relationship with the RB gene mutation. Anticancer Res. 1998;18(4A):2317–21.
Motokura T, Arnold A. Cyclin D and oncogenesis. Curr Opin Genet Dev. 1993;3(1):5–10.
Zhang SY, Caamano J, Cooper F, Guo X, Klein-Szanto AJ. Immunohistochemistry of cyclin D1 in human breast cancer. Am J Clin Pathol. 1994;102(5):695–8.
Naitoh H, Shibata J, Kawaguchi A, Kodama M, Hattori T. Overexpression and localization of cyclin D1 mRNA and antigen in esophageal cancer. Am J Clin Pathol. 1995;146(5):1161–9.
Betticher DC, Heighway J, Hasleton PS, Altermatt HJ, Ryder WD, Cerny T, et al. Prognostic significance of CCND1 (cyclin D1) overexpression in primary resected non-small-cell lung cancer. Br J Cancer. 1996;73(3):294–300.
Lin HS, Berry GJ, Sun Z, Fee WE Jr. Cyclin D1 and p16 expression in recurrent nasopharyngeal carcinoma. World J Surg Oncol. 2006;4:62.
Gimenez-Conti IB, Collet AM, Lanfranchi H, Itoiz ME, Luna M, Xu HJ, et al. p53, Rb, and cyclin D1 expression in human oral verrucous carcinomas. Cancer. 1996;78(1):17–23.
Michalides RJ, van Veelen NM, Kristel PM, Hart AA, Loftus BM, Hilgers FJ, et al. Overexpression of cyclin D1 indicates a poor prognosis in squamous cell carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg. 1997;123(5):497–502.
Staibano S, Lo Muzio L, Pannone G, Mezza E, Argenziano G, Vetrani A, et al. DNA ploidy and cyclin D1 expression in basal cell carcinoma of the head and neck. Am J Clin Pathol. 2001;115(6):805–13.
Zhang YJ, Jiang W, Chen CJ, Lee CS, Kahn SM, Santella RM, et al. Amplification and overexpression of cyclin D1 in human hepatocellular carcinoma. Biochem Biophys Res Commun. 1993;196(2):1010–6.
Nai G, Marques M. Role of ROC1 protein in the control of cyclin D1 protein expression in skin melanomas. Pathol Res Pract. 2011;207(3):174–81.
Graf F, Koehler L, Kniess T, Wuest F, Mosch B, Pietzsch J. Cell cycle regulating kinase Cdk4 as a potential target for tumor cell treatment and tumor imaging. J Oncol. 2009;2009(17):106378.
Malumbres M, Barbacid M. Is cyclin D1-CDK4 kinase a bona fide cancer target? Cancer Cell. 2006;9(1):2–4.
Musgrove EA, Caldon CE, Barraclough J, Stone A, Sutherland RL. Cyclin D as a therapeutic target in cancer. Nat Rev Cancer. 2011;11(8):558–72.
Kim JK, Diehl JA. Nuclear cyclin D1: an oncogenic driver in human cancer. J Cell Physiol. 2009;220(2):292–6.
El-Serag HB. Hepatocellular carcinoma: an epidemiologic view. J Clin Gastroenterol. 2002;35(5 Suppl 2):S72–8.
El-Serag HB, Rudolph KL. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology. 2007;132(7):2557–76.
Jain S, Singhal S, Lee P, Xu R. Molecular genetics of hepatocellular neoplasia. Am J Transl Res. 2010;2(1):105–18.
Lin TY, Lee CS, Chen KM, Chen CC. Role of surgery in the treatment of primary carcinoma of the liver: a 31-year experience. Br J Surg. 1987;74(9):839–42.
Sakamoto M. Early HCC: diagnosis and molecular markers. J Gastroenterol. 2009;44(Suppl 19):108–11.
Hirohashi S, Ishak KG, KojiroM, Puig PL, Wanless IR, Fischer HP, TheiseND, SakamotoM, Tsukuma H. Hepatocellular carcinoma. In: Hamilton SR, Aaltonen LA, editors. Pathology and genetics of tumours of the digestive system. Lyon: IARCPress; 2000. p. 159–72.
Lippincott-Raven P. Digestive system. In: American Joint Committee on Cancer, editor. AJCC cancer staging manual. New York: Springer; 2009. p. 191–200.
Yeh KT, Yang MY, Liu TC, Chen JC, Chan WL, Lin SF, et al. Abnormal expression of period 1 (PER1) in endometrial carcinoma. J Pathol. 2005;206(1):111–20.
Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008;3(6):1101–8.
Chang YS, Yeh KT, Yang MY, Liu TC, Lin SF, Chan WL, et al. Abnormal expression of JUNB gene in hepatocellular carcinoma. Oncol Rep. 2005;13(3):433–8.
Lu JW, Chang JG, Yeh KT, Chen RM, Tsai JJ, Hu RM. Decreased expression of p39 is associated with a poor prognosis in human hepatocellular carcinoma. Med Oncol. 2011;28(Suppl 1):S239–45.
Lu JW, Chang JG, Yeh KT, Chen RM, Tsai JJ, Hu RM. Overexpression of Thy1/CD90 in human hepatocellular carcinoma is associated with HBV infection and poor prognosis. Acta Histochem. 2011;113:833–8.
Mantel N. Evaluation of survival data and two new rank order statistics arising in its consideration. Cancer Chemother Rep. 1966;50(3):163–70.
Xu XR, Huang J, Xu ZG, Qian BZ, Zhu ZD, Yan Q, et al. Insight into hepatocellular carcinogenesis at transcriptome level by comparing gene expression profiles of hepatocellular carcinoma with those of corresponding noncancerous liver. Proc Natl Acad Sci USA. 2001;98(26):15089–94.
Chen X, Cheung ST, So S, Fan ST, Barry C, Higgins J, et al. Gene expression patterns in human liver cancers. Mol Biol Cell. 2002;13(6):1929–39.
Dawson S, Apcher S, Mee M, Higashitsuji H, Baker R, Uhle S, et al. Gankyrin is an ankyrin-repeat oncoprotein that interacts with CDK4 kinase and the S6 ATPase of the 26 S proteasome. J Biol Chem. 2002;277(13):10893–902.
Azechi H, Nishida N, Fukuda Y, Nishimura T, Minata M, Katsuma H, et al. Disruption of the p16/cyclin D1/retinoblastoma protein pathway in the majority of human hepatocellular carcinomas. Oncology. 2001;60(4):346–54.
Higashitsuji H, Liu Y, Mayer RJ, Fujita J. The oncoprotein gankyrin negatively regulates both p53 and RB by enhancing proteasomal degradation. Cell Cycle. 2005;4(10):1335–7.
Huang SF, Cheng SD, Chuang WY, Chen IH, Liao CT, Wang HM, et al. Cyclin D1 overexpression and poor clinical outcomes in Taiwanese oral cavity squamous cell carcinoma. World J Surg Oncol. 2012;10(40):40.
Mahdey HM, Ramanathan A, Ismail SM, Abraham MT, Jamaluddin M, Zain RB. Cyclin D1 amplification in tongue and cheek squamous cell carcinoma. Asian Pac J Cancer Prev. 2011;12(9):2199–204.
Zhao J, Li L, Wei S, Gao Y, Chen Y, Wang G, et al. Clinicopathological and prognostic role of cyclin D1 in esophageal squamous cell carcinoma: a meta-analysis. Dis Esophagus. 2012;25(6):520–6.
Bilyk OO, Pande NT, Buchynska LG. Analysis of p53, p16(INK4a), pRb and cyclin D1 expression and human papillomavirus in primary ovarian serous carcinomas. Exp Oncol. 2011;33(3):150–6.
Hashimoto T, Yanaihara N, Okamoto A, Nikaido T, Saito M, Takakura S, et al. Cyclin D1 predicts the prognosis of advanced serous ovarian cancer. Exp Ther Med. 2011;2(2):213–9.
Choschzick M, Hess S, Tennstedt P, Holst F, Bohlken H, Gieseking F, et al. Role of cyclin D1 amplification and expression in vulvar carcinomas. Hum Pathol. 2012;43(9):1386–93.
Gunia S, Kakies C, Erbersdobler A, Hakenberg OW, Koch S, May M. Expression of p53, p21 and cyclin D1 in penile cancer: p53 predicts poor prognosis. J Clin Pathol. 2012;65(3):232–6.
Jang KY, Kim YN, Bae JS, Chung MJ, Moon WS, Kang MJ, et al. Expression of cyclin D1 is associated with beta-catenin expression and correlates with good prognosis in colorectal adenocarcinoma. Transl Oncol. 2012;5(5):370–8.
Lehn S, Tobin NP, Berglund P, Nilsson K, Sims AH, Jirstrom K, et al. Down-regulation of the oncogene cyclin D1 increases migratory capacity in breast cancer and is linked to unfavorable prognostic features. Am J Pathol. 2010;177(6):2886–97.
Tobin NP, Sims AH, Lundgren KL, Lehn S, Landberg G. Cyclin D1, Id1 and EMT in breast cancer. BMC Cancer. 2011;11(417):417.
Peng SY, Chou SP, Hsu HC. Association of downregulation of cyclin D1 and of overexpression of cyclin E with p53 mutation, high tumor grade and poor prognosis in hepatocellular carcinoma. J Hepatol. 1998;29(2):281–9.
Liu F, Lee WH. CtIP activates its own and cyclin D1 promoters via the E2F/RB pathway during G1/S progression. Mol Cell Biol. 2006;26(8):3124–34.
Muller H, Lukas J, Schneider A, Warthoe P, Bartek J, Eilers M, et al. Cyclin D1 expression is regulated by the retinoblastoma protein. Proc Natl Acad Sci USA. 1994;91(8):2945–9.
Nishida N, Fukuda Y, Komeda T, Kita R, Sando T, Furukawa M, et al. Amplification and overexpression of the cyclin D1 gene in aggressive human hepatocellular carcinoma. Cancer Res. 1994;54(12):3107–10.
Ito Y, Matsuura N, Sakon M, Miyoshi E, Noda K, Takeda T, et al. Expression and prognostic roles of the G1-S modulators in hepatocellular carcinoma: p27 independently predicts the recurrence. Hepatology. 1999;30(1):90–9.
Joo M, Kang YK, Kim MR, Lee HK, Jang JJ. Cyclin D1 overexpression in hepatocellular carcinoma. Liver. 2001;21(2):89–95.
Lukas J, Pagano M, Staskova Z, Draetta G, Bartek J. Cyclin D1 protein oscillates and is essential for cell cycle progression in human tumour cell lines. Oncogene. 1994;9(3):707–18.
Acknowledgments
This study was supported by National Science Council of Taiwan Grant Nos. NSC-95-2745-E-468-005-URD for RM Hu, NSC-95-2745-E-468-001-URD for JP Tsai, and NSC-94-2745-E-468-003-URD for RM Chen.
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The authors declare that they have no conflict of interest relating to the publication of this manuscript.
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Jeng-Wei Lu and Yueh-Min Lin contributed equally to this work.
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Lu, JW., Lin, YM., Chang, JG. et al. Clinical implications of deregulated CDK4 and Cyclin D1 expression in patients with human hepatocellular carcinoma. Med Oncol 30, 379 (2013). https://doi.org/10.1007/s12032-012-0379-5
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DOI: https://doi.org/10.1007/s12032-012-0379-5