Skip to main content

Advertisement

SpringerLink
Log in

Clinicopathological significance of RASSF1A reduced expression and hypermethylation in hepatocellular carcinoma

  • Original Article
  • Published:
Hepatology International Aims and scope Submit manuscript

Abstract

Purpose

Protein downregulation and hypermethylation of Ras association domain family 1A (RASSF1A) has been recognized as an important early event in different classes of carcinogenesis, but clinicopathological significance of RASSF1A protein expression and methylation in hepatocellular carcinoma (HCC) remains largely unknown. The aim of the study was to investigate the expression of RASSF1A protein and methylation in HCC and their clinical significance.

Methods

Immunohistochemistry was employed to detect the expression of RASSF1A proteins in liver tissue microarrays. Aberrant promoter hypermethylation of RASSF1A was investigated in DNA from HCC, matching noncancerous tissues and serum of 35 HCC patients by methylation-specific PCR.

Results

RASSF1A protein expression in HCC was significantly lower than that in noncancerous (p = 0.015) and paracancerous tissues (p = 0.017). In addition, reduced RASSF1A protein expression is related to TNM stage, metastasis, α-fetoprotein, portal vein embolus, capsular infiltration, and multiple tumor nodes. Furthermore, RASSF1A promoter methylation in HCC was significantly higher than that in noncancerous liver tissues (p < 0.05). Meanwhile, RASSF1A promoter hypermethylation was detected in 14 in the serum DNA from HCC patients, whereas no hypermethylation was detected in the normal controls. Hypermethylation of RASSF1A in HCC serum and tissues was negatively correlated with the expression of RASSF1A protein expression (p < 0.05).

Conclusions

The loss or abnormal protein downregulation and the promoter hypermethylation of RASSF1A could play important roles in the tumorigenesis development and metastases of HCC. The detection of the promoter hypermethylation of RASSF1A in serum DNA could be a valuable biomarker for early-stage diagnosis in populations at high risk of HCC.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Orito E, Mizokami M. Differences of HBV genotypes and hepatocellular carcinoma in Asian countries. Hepatol Res 2007;37:S33–35

    Article  CAS  PubMed  Google Scholar 

  2. Nagai H, Sumino Y. Therapeutic strategy of advanced hepatocellular carcinoma by using combined intra-arterial chemotherapy. Recent Pat Anticancer Drug Discov 2008;3:220–226

    Article  CAS  PubMed  Google Scholar 

  3. Dammann R, Li C, Yoon JH, Chin PL, Bates S, Pfeifer GP. Epigenetic inactivation of a RAS association domain family protein from the lung tumour suppressor locus 3p21.3. Nat Genet 2000;25:315–319

    Article  CAS  PubMed  Google Scholar 

  4. Tezval H, Merseburger AS, Matuschek I, Machtens S, Kuczyk MA, Serth J. RASSF1A protein expression and correlation with clinicopathological parameters in renal cell carcinoma. BMC Urol 2008;8:12

    Article  PubMed  CAS  Google Scholar 

  5. Ghosh S, Ghosh A, Maiti GP, Alam N, Roy A, Roy B, et al. Alterations of 3p21.31 tumor suppressor genes in head and neck squamous cell carcinoma: correlation with progression and prognosis. Int J Cancer 2008;123:2594–2604

    Article  CAS  PubMed  Google Scholar 

  6. Lee JS, Fackler MJ, Teo WW, Lee JH, Choi C, Park MH, et al. Quantitative promoter hypermethylation profiles of ductal carcinoma in situ in North American and Korean women: potential applications for diagnosis. Cancer Biol Ther 2008;7:1398–1406

    Article  CAS  PubMed  Google Scholar 

  7. Pallarés J, Velasco A, Eritja N, Santacana M, Dolcet X, Cuatrecasas M, et al. Promoter hypermethylation and reduced expression of RASSF1A are frequent molecular alterations of endometrial carcinoma. Mod Pathol 2008;21:691–699

    Article  PubMed  CAS  Google Scholar 

  8. Lo PH, Xie D, Chan KC, Xu FP, Kuzmin I, Lerman MI, et al. Reduced expression of RASSF1A in esophageal and nasopharyngeal carcinomas significantly correlates with tumor stage. Cancer Lett 2007;257:199–205

    Article  CAS  PubMed  Google Scholar 

  9. Ye M, Xia B, Guo Q, Zhou F, Zhang X. Association of diminished expression of RASSF1A with promoter methylation in primary gastric cancer from patients of central China. BMC Cancer 2007;7:120

    Article  PubMed  CAS  Google Scholar 

  10. Sung JS, Han SG, Whang YM, Shin ES, Lee JW, Lee HJ, et al. Putative association of the single nucleotide polymorphisms in RASSF1A promoter with Korean lung cancer. Lung Cancer 2008;61:301–308

    Article  PubMed  Google Scholar 

  11. Kee SK, Lee JY, Kim MJ, Lee SM, Jung YW, Kim YJ, et al. Hypermethylation of the Ras association domain family 1A (RASSF1A) gene in gallbladder cancer. Mol Cells 2007;24:364–371

    CAS  PubMed  Google Scholar 

  12. Li Y, Wei Q, Cao F, Cao X. Expression and promoter hypermethylation of the RASSF1A gene in sporadic breast cancers in Chinese women. Oncol Rep 2008;19:1149–1153

    CAS  PubMed  Google Scholar 

  13. Peters I, Vaske B, Albrecht K, Kuczyk MA, Jonas U, Serth J. Adiposity and age are statistically related to enhanced RASSF1A tumor suppressor gene promoter hypermethylation in normal autopsy kidney tissue. Cancer Epidemiol Biomarkers Prev 2007;16:2526–2532

    Article  CAS  PubMed  Google Scholar 

  14. Jagadeesh S, Sinha S, Pal BC, Bhattacharya S, Banerjee PP. Mahanine reverses an epigenetically silenced tumor suppressor gene RASSF1A in human prostate cancer cells. Biochem Biophys Res Commun 2007;362:212–217

    Article  CAS  PubMed  Google Scholar 

  15. Hu J, Li H, Shi T, Ma X, Wang B, Xu H, et al. Relationship between the expression of RASSF1A protein and promoter hypermethylation of RASSF1A gene in bladder tumor. J Huazhong Univ Sci Technol Med Sci 2008;28:182–184

    Article  CAS  PubMed  Google Scholar 

  16. Chen YJ, Tang QB, Zou SQ. Inactivation of RASSF1A, the tumor suppressor gene at 3p21.3 in extrahepatic cholangiocarcinoma. World J Gastroenterol 2005;11:1333–1338

    CAS  PubMed  Google Scholar 

  17. Wang YC, Yu ZH, Liu C, Xu LZ, Yu W, Lu J, et al. Detection of RASSF1A promoter hypermethylation in plasma from gastric and colorectal adenocarcinoma patients. World J Gastroenterol 2008;14:3074–3080

    Article  CAS  PubMed  Google Scholar 

  18. Yeo W, Wong N, Wong WL, Lai PB, Zhong S, Johnson PJ. High frequency of promoter hypermethylation of RASSF1A in tumor and serum of patients with hepatocellular carcinoma. Liver Int 2005;25:266–272

    Article  CAS  PubMed  Google Scholar 

  19. Zhang YJ, Wu HC, Shen J, Ahsan H, Tsai WY, Yang HI, et al. Predicting hepatocellular carcinoma by detection of aberrant promoter hypermethylation in serum DNA. Clin Cancer Res 2007;13:2378–2384

    Article  CAS  PubMed  Google Scholar 

  20. Zhang C, Li Z, Cheng Y, Jia F, Li R, Wu M, et al. CpG island methylator phenotype association with elevated serum alpha-fetoprotein level in hepatocellular carcinoma. Clin Cancer Res 2007;13:944–952

    Article  CAS  PubMed  Google Scholar 

  21. Di Gioia S, Bianchi P, Destro A, Grizzi F, Malesci A, Laghi L, et al. Quantitative evaluation of RASSF1A methylation in the non-lesional, regenerative and neoplastic liver. BMC Cancer 2006;6:89

    Article  PubMed  CAS  Google Scholar 

  22. Tischoff I, Markwarth A, Witzigmann H, Uhlmann D, Hauss J, Mirmohammadsadegh A, et al. Allele loss and epigenetic inactivation of 3p21.3 in malignant liver tumors. Int J Cancer 2005;115:684–689

    Article  CAS  PubMed  Google Scholar 

  23. Schagdarsurengin U, Wilkens L, Steinemann D, Flemming P, Kreipe HH, Pfeifer GP, et al. Frequent epigenetic inactivation of the RASSF1A gene in hepatocellular carcinoma. Oncogene 2003;22:1866–1871

    Article  CAS  PubMed  Google Scholar 

  24. Kononen J, Bubendorf L, Kallioniemi A, Bärlund M, Schraml P, Leighton S, et al. Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 1998;4:844–847

    Article  CAS  PubMed  Google Scholar 

  25. Brennan DJ, Kelly C, Rexhepaj E, Dervan PA, Duffy MJ, Gallagher WM. Contribution of DNA and tissue microarray technology to the identification and validation of biomarkers and personalised medicine in breast cancer. Cancer Genomics Proteomics 2007;4:121–134

    CAS  PubMed  Google Scholar 

  26. Lemmer ER, Friedman SL, Llovet JM. Molecular diagnosis of chronic liver disease and hepatocellular carcinoma: the potential of gene expression profiling. Semin Liver Dis 2006;26:373–384

    Article  CAS  PubMed  Google Scholar 

  27. Ko SS, Na YS, Yoon CS, Park JY, Kim HS, Hur MH, et al. The significance of c-erbB-2 overexpression and p53 expression in patients with axillary lymph node-negative breast cancer: a tissue microarray study. Int J Surg Pathol 2007;15:98–109

    Article  CAS  PubMed  Google Scholar 

  28. Xue WJ, Li C, Zhou XJ, Guan HG, Qin L, Li P, et al. RASSF1A expression inhibits the growth of hepatocellular carcinoma from Qidong County. J Gastroenterol Hepatol 2008;23:1448–1458

    Article  CAS  PubMed  Google Scholar 

  29. Kwong J, Lo KW, To KF, Teo PM, Johnson PJ, Huang DP. Promoter hypermethylation of multiple genes in nasopharyngeal carcinoma. Clin Cancer Res 2002;8:131–137

    CAS  PubMed  Google Scholar 

  30. Hsu HS, Chen TP, Hung CH, Wen CK, Lin RK, Lee HC, et al. Characterization of a multiple epigenetic marker panel for lung cancer detection and risk assessment in serum. Cancer 2007;110:2019–2026

    Article  CAS  PubMed  Google Scholar 

  31. Chan MW, Chan LW, Tang NL, Lo KW, Tong JH, Chan AW, et al. Frequent hypermethylation of promoter region of RASSF1A in tumor tissues and voided urine of urinary bladder cancer patients. Int J Cancer 2003;104:611–616

    Article  CAS  PubMed  Google Scholar 

  32. Dulaimi E, Hillinck J, Ibanez de Caceres I, Al-Saleem T, Cairns P. Tumor suppressor gene promoter hypermethylation in serum of breast cancer patients. Clin Cancer Res 2004;10:6189–6193

    Article  CAS  PubMed  Google Scholar 

  33. Shukla S, Mirza S, Sharma G, Parshad R, Gupta SD, Ralhan R. Detection of RASSF1A and RARbeta hypermethylation in serum DNA from breast cancer patients. Epigenetics 2006;1:88–93

    PubMed  Google Scholar 

  34. Wong TS, Kwong DL, Sham JS, Wei WI, Kwong YL, Yuen AP. Quantitative serum hypermethylated DNA markers of undifferentiated nasopharyngeal carcinoma. Clin Cancer Res 2004;10:2401–2406

    Article  CAS  PubMed  Google Scholar 

  35. Bolondi L. Screening for hepatocellular carcinoma in cirrhosis. J Hepatol 2003;39:1076–1084

    Article  PubMed  Google Scholar 

  36. Sherman M. Alphafetoprotein: an obituary. J Hepatol 2001;34:603–605

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was performed in People’s Republic of China. The study was supported by Medical Major Scientific Research Foundation of Guangxi (Zhong200513). The authors thank Dr. Ping Li for her immunodetection work and Meinhart Francois from Belgium for his useful suggestions in the manuscript preparation.

Author information

Authors and Affiliations

  1. Department of Epidemiology, Guangxi Medical University, Nanning, 530021, Guangxi Zhuang Autonomous Region, People’s Republic of China

    Lang Hu, Hongping Yu & Xiaoqiang Qiu

  2. Department of Pathology, Guangxi Medical University, Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People’s Republic of China

    Gang Chen

Authors
  1. Lang Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hongping Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaoqiang Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiaoqiang Qiu.

Additional information

L. Hu and G. Chen contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hu, L., Chen, G., Yu, H. et al. Clinicopathological significance of RASSF1A reduced expression and hypermethylation in hepatocellular carcinoma. Hepatol Int 4, 423–432 (2010). https://doi.org/10.1007/s12072-010-9164-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12072-010-9164-8

Keywords

Search

Navigation