Cyclin D1, Retinoblastoma and p16 Protein Expression in Carcinoma of the Gallbladder

  • Srivastava, Vineeta (Department of Surgical Oncology and Pathology, Institute of Medical Sciences, Banaras Hindu University) ;
  • Patel, Brijesh (Department of Surgical Oncology and Pathology, Institute of Medical Sciences, Banaras Hindu University) ;
  • Kumar, Mohan (Department of Surgical Oncology and Pathology, Institute of Medical Sciences, Banaras Hindu University) ;
  • Shukla, Mridula (Department of Surgical Oncology and Pathology, Institute of Medical Sciences, Banaras Hindu University) ;
  • Pandey, Manoj (Department of Surgical Oncology and Pathology, Institute of Medical Sciences, Banaras Hindu University)
  • Published : 2013.05.30


Background: Cancer of the gallbladder is a relatively rare neoplasm with a poor prognosis. The exact mechanisms of its genesis are not known and very little information is available on molecular events leading to labeling this as an orphan cancer. Materials and Methods: In this prospective case control study we evaluated the expression of p16, pRb and cyclin D1 by immunohistochemistry to study the G1-S cell-cycle check point and its possible role in gallbladder carcinogenesis. A total of 25 patients with gallbladder carcinoma (group I), 25 with cholelithiasis (group II) and 10 normal controls. were enrolled Results: Cyclin D1 expression was seen in 10 (40%) patients each with carcinoma and cholelithiasis while only in 2 (20%) of the normal gallbladders but differences were not statistically significant (p value=0.488). p16 was expressed in 12% patients of carcinoma of the gallbladder and 28% of cholelithiasis, however this difference was not statistically significant (p value=0.095). Retinoblastoma protein was found to be expressed in 50% of normal gallbladders and 6 (24%) of carcinoma and 8 (32%) of gallstones. The present study failed to demonstrate any conclusive role of cyclin D1/RB/ p16 pathway in carcinoma of the gallbladder. Conclusions: The positive relation observed between tumor metastasis and cyclinD1 expression and p16 with nodal metastasis suggested that higher cyclin D1/p16 expression may act as a predictive biomarker for aggressive behavior of gallbladder malignancies.


  1. Beasley MB, Lantuejoul S, Abbondanzo S, et al (2003). the p16/cyclin D1/ Rb pathway in neuroendocrine tumors of the lung. Hum Pathol, 34, 136-42.
  2. Brantley MA Jr, Harbour JW (2000). Deregulation of the Rb and p53 pathwaysin uveal melanoma. Am J Pathol, 157, 795-1801.
  3. Cho NH, Kim YT, Kim JW (2002). Alteration of cell-cycle in cervical tumor associated with human papilloma virus: cyclin dependent kinase inhibitor. Yonsei Med J, 43, 722-8.
  4. Choi HJ, Yun SS, Kim HJ, Choi JH (2010). Expression of p16 protein in gallbladder carcinoma and its precancerous conditions. Hepatogastroenterology, 57, 18-21.
  5. Cubertafond P, Gainant A, Cucchiaro G (1994). Surgical treatment of 724 carcinoma of the gallbladder. Results of the French surgical association survey. Ann Surg, 219, 275-80.
  6. Das SK, Hashimoto T, Schimizu K, et al (2005). Fucoxanthin induces cell cycle arrest at G0/G1 phase in human colon carcinoma cells through up-regulation of p21WAF1/Cip1. Biochim Biophys Acta, 1726, 328-35.
  7. Dasgupta P, BettsV, Rastogi S, et al (2004). Direct binding of apoptosis signal-regulating kinase 1 to retinoblastoma protein: novel links between apoptotic signaling and cell cyclr machinery. J Biol Chem, 279, 762-9.
  8. Fan YZ, Zhang JT, Yang HC, Yang YQ (2002). Expression of MMP-2, TIMP-2 protein and the ratio of MMP-2/ TIMP-2 in gallbladder carcinoma and their significance. World J Gastrointerol, 8, 1138-43.
  9. Feng Z, Chen J, Wei H, et al (2011). The risk factor of gallbladder cancer: hyperplasia of mucous epithelium caused by gallstones associates with p16/CyclinD1/CDK4 pathway Exp Mol Pathol, , 569-77.
  10. Fleming ID, Cooper JS, Henson DE, et al (1997). AJCC cancer staging manual. 5th ed. JB Lippincott, Philadelphia 1997.
  11. Gillett C, Fantl V, Smith R, et al (1994). Amplification and overexpression of Cylin D1 in breast cancer detected by immunohistochemical staining. Cancer Res, 54, 1812-7.
  12. Gregorc V, Ludovini V, Pistola L, et al (2003). Relevance of p53, bcl-2 and Rb expression on resistance to cisplatin-based chemotherapy in advanced non-small cell lung cancer. Lung Cancer, 39, 41-8.
  13. Guner D, Sturm I, Hemmanti P, et al (2003). Multigene analysis of Rb pathway and apoptosis control in esophageal squamous cell carcinoma identifies patients with good prognosis. Int J Cancer, 103, 445-54.
  14. Hartwell LH, Kastan MB (1994). Cell cycle control and cancer. Science, 266, 1821-8.
  15. Hawang CF, Cho CL, Huang CC, et al (2002). Loss of cyclin D1 and p16 expression correlates with local recurrence of nasopharyngeal carcinoma following radiotherapy. Ann Oncol, 13, 1246-51.
  16. Ito H, Matros E, Brooks DC, et al (2004). Treatment outcomes associated with surgery for gallbladder cancer: a 20-year experience. J Gastrointest Surg, 8, 183-90.
  17. Jemal A, Murray T, Ward E, et al (2005). Cancer Statics. CA Cancer J Clin, 55, 10-30.
  18. Kamb A, Gruis NA, Weaver-Feldhaus J, et al (1994). A cell cycle regulator potentially involved in genesis of many tumor types. Science, 264, 436-40.
  19. Kang YK, Kim WH, Jang JJ (2002). Expression of G1/S modulators (p53, p16, p27, cyclin D1, Rb) and Smad4/Dpc4 in intrahepatic cholangio carcinoma. Hum Pathol, 33, 877-83.
  20. Kato J, Matsushime H, Heibert SW, EwenME, Sherr CJ (1993). Direct binding of cyclin D to the retinoblastoma gene product (pRB) and pRb phosphorylation by the cyclin D ? Dependent kinase CDK4. Genes Dev, 7, 331-42.
  21. Lazcano-ponce EC, Miqual JF, Munoz N, et al (2001). Epidemiology and moleecular pathologyof gallbladder cancer. CA Cancer J Clin, 51, 349-64.
  22. Lerma E, Esteler M, Harman JC, Prat J (2002). Alteration of p16/Rb/cyclin D1 pathway in vulvular carcinoma, vulvular intraepithelial neoplasia and lochen sclerosis. Hum Pathol, 33, 1020-5.
  23. Levy AD, Murakata LA, Rohrmann CA (2001). Gallbladder carcinoma: radiologic- pathologic correlation. Radiographics, 21, 295-314.
  24. Li W, Sanki A, Karim RZ, et al (2006). The role of cell cycle regulatory proteins in the pathogenesis of melanoma. Pathology, 38, 287-301.
  25. Lynch BC, Lathrop SL, Ye D, Ma TY, Cerilli LA (2008). Expression of the p16(INK4a) gene product in premalignant and malignant epithelial lesions of the gallbladder. Ann Diagn Pathol, 12, 161-4.
  26. Ma HB, Hu HT, Di ZL, et al (2005). Association of cyclin D1, p16 1nd retinoblastoma protein expressions with prognosis and metastasis of gallbladder carcinoma. World J Gastroenterol, 11, 744-7.
  27. Michalides R (2002). Cell cycle regulators: role in etiology, prognosis and treatment in cancer. Ann Oncol, 13, 39.
  28. Misra S, Chaturvedi A, Misra NC, Sharma ID (2003). Carcinoma of the gallbladder. Lancet Oncol, 4, 167-76.
  29. Murphree AL, Benedict WF (1984). Retinoblastoma: clues to human oncogenesis. Science, 223, 1028-33.
  30. Offerhaus GTA (2002). Tumors of the gallbladder, extrahepatic bile ducts and ampulla of vater. atlas of tumor pathology. J Clin Pathol, 54, 816.
  31. Pan MH, Chen WJ, Lin-Shiau SY, Ho CT, Lin JK (2002). Tangeretin induces cell-cycle G1 arrest through inhibiting cyclin-dependent kinase 2 and 4 activities as well as elevating Cdk inhibitors p21 and p27 in human colorectal carcinoma cells. Carcinogenesis, 23, 1677-84.
  32. Pei XH, Xiong Y (2005). Biochemical and cellular mechanisms of mammalian CDK inhitors: a few unresolved issue. Oncogene, 24, 2787-95.
  33. Pandey M, Pathak AK, Singh S, Gautam A, Shukla VK (2001). Carcinoma of the gallbladder: a retrospective review of 99 cases. Dig Dis Sci, 46, 1145-51.
  34. Pandey M (2003). Risk factors for gallbladder cancer a reappraisal. Eur J Cancer Prev, 12, 15-24.
  35. Quan Zw, Wu K, Wang J, et al (2001). Association of p53, p16, and vascular endothelial growth factor protein expression with the prognosis and metastasis of gallbladder cancer. J Am Coll Surg, 193, 380-3.
  36. Raghvan D (2003). Molecular targeting and pharmacogenomics in the management of advanced bladder cancer. Cancer, 97, 2083-9.
  37. Rifatbegovic Z, Mesic D, Ljuca F, Zildzic M, Morankic M (2007). Incidence and surgical treatment of cancer in gallbladder. Med Arh, 61, 30-3.
  38. Russell A, Thompson MA, Hendley J, et al (1999). Cyclin D1 and D3 associate with the SCF complex and are coordinately elevated in breast cancer. Oncogene, 18, 1983-91.
  39. Said K, Glaumann H, Bjornstedt M, Bergquist A (2012). The value of thioredoxin family proteins and proliferation markers in dysplastic and malignant gallbladders in patients with primary sclerosing cholangitis. Dig Dis Sci, 57, 1163-70.
  40. Sauter ER, Yeo UC, Von Stemm A, et al (2002). Cycin D1 is a candidate oncogene in cutaneous melanoma. Cancer Res, 62, 3200-6.
  41. Sdek P, Zhang Z, Coa J (2002). Influence of HPV 16 on the expression of p16/ cyclin D1/ Rb in oro epithelial cancer. Zhongua Kouqiang Yixue Zazhi, 16, 84-6.
  42. Serrano M, Hannon GJ, Beach D (1993). A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature, 366, 704-7.
  43. Sherr CJ (1996). Cancer cell cycles. Science, 274, 1672-7.
  44. Shi YZ, Hui AM, Li X, Takayama T, Makuuchi M (2000). Overexpression of retinoblastoma protein predicts decreased survival and correlates with loss of p161INK4 protein in gallbladder carcinomas. Clin Cancer Res, 6, 4096-100.
  45. Tadokoro H, Shigihara T, Ikeda T, Takase M, Suyama M (2007). Two distinct pathways of p16 gene inactivation in gallbladder cancer. World J Gastroenterol, 13, 6396-403.
  46. Weinberg RA (1995). The retinoblastoma protein and cell cycle control. Cell, 81, 323-30.
  47. Xuan YH, Choi YL, Shin YK, et al (2005). An immunohistochemical study of the expression of cell-cycleregulated proteins p53, cyclin D1, RB, p27, Ki67 and MSH2 in gallbladder carcinoma and its precursor lesions. Histol Histopathol, 20, 59-66.
  48. Yoo J, Park SY, Kang SJ, Shim SI, Kim BK (2002). Altered expression of G1 regulatory proteins in human soft tissue sarcomas. Arch Pathol Lab Med, 126, 567-73.
  49. Zhang R, Zhang JJ, He ZC, Cheng SJ, Gao YN (2003). Research advances on bladder cancer associated genes. Aizheng, 22, 104-7.

Cited by

  1. High Frequency of TP53 but not K-ras Gene Mutations in Bolivian Patients with Gallbladder Cancer vol.15, pp.13, 2014,
  2. Clinical Efficacy and Prognostic Factors of Chemoreduction Combined with Topical Treatment for Advanced Intraocular Retinoblastoma vol.15, pp.18, 2014,
  3. Role of the prostaglandin E2/E-prostanoid 2 receptor signalling pathway in TGFβ-induced mice mesangial cell damage vol.34, pp.6, 2014,
  4. Expression in Squamous Cell Carcinomas of the Vagina and the Vulva in Tunisian Women vol.15, pp.24, 2015,
  5. Elevated GRP78 expression is associated with poor prognosis in patients with pancreatic cancer vol.5, pp.1, 2015,
  6. Surgical treatment of gallbladder carcinoma: a critical review vol.67, pp.4, 2015,
  7. Role of the ER stress in prostaglandin E2/E-prostanoid 2 receptor involved TGF-β1-induced mice mesangial cell injury vol.411, pp.1-2, 2016,