Clinical & Experimental Metastasis

, Volume 26, Issue 7, pp 839–848

Prognostic factors and recurrence of hepatitis B-related hepatocellular carcinoma after argon-helium cryoablation: a prospective study

  • Chunping Wang
  • Yinying Lu
  • Yan Chen
  • Yongyi Feng
  • Linjing An
  • Xinzhen Wang
  • Shuhui Su
  • Wenlin Bai
  • Lin Zhou
  • Yongping Yang
  • Dongping Xu
Research paper


To determine the long-term prognosis of hepatocellular carcinoma (HCC) after argon-helium cryoablation and identify the risk factors that predict metastasis and recurrence. A total of 156 patients with hepatitis B-related HCC less than 5 cm in diameter who underwent curative cryoablation were followed up prospectively for tumor metastasis and recurrence. Immunohistochemistry was used to analyze the expression of vascular endothelial growth factor (VEGF). HBV basal core promoter (BCP) and precore mutations were detected by DNA sequence analysis. Post-treatment prognostic factors influencing survival, tumor metastasis and recurrence were assessed by univariate and multivariate analyses. The variables included the expression of VEGF in HCC tissues, clinical and pathologic characteristics of patients, and HBV features (HBV DNA level, HBV genotype, BCP mutation). The median follow-up period of the 156 patients was 37 months (range 8–48 months). The 1-, 2-, and 3-year overall survival rates were 92, 82 and 64%, respectively. The 1-, 2-, and 3-year recurrence-free survival rates were 72, 56 and 43%, respectively. Eighty-five patients (54.5%) had tumor recurrence or metastasis. The multivariate analysis showed that Child-Pugh class and the expression of VEGF in HCC tissues could be used as independent prognostic factors for overall survival. Meanwhile, the expression of VEGF in HCC tissues and HBV BCP mutations were found to be independent prognostic factors for recurrence-free survival. Strong expression of VEGF in HCC tissues and HBV BCP mutations are important risk predictors for recurrence or metastasis of HCC smaller than 5 cm in diameter.


Hepatocellular carcinoma Cryoablation Prognostic factor Recurrence Vascular endothelial growth factor HBV basal core promoter 



Computed tomography


Magnetic resonance imaging


Polymerase chain reaction


  1. 1.
    Parkin DM (2001) Global cancer statistics in the year 2000. Lancet Oncol 2(9):533–543 [PMID: 11905707]CrossRefPubMedGoogle Scholar
  2. 2.
    Yang HI, Lu SN, Liaw YF, You SL, Sun CA, Wang LY, Hsiao CK, Chen PJ, Chen DS, Chen CJ (2002) Hepatitis B e antigen and the risk of hepatocellular carcinoma. N Engl J Med 347(3):168–174 [PMID: 12124405]CrossRefPubMedGoogle Scholar
  3. 3.
    Poon RT, Fan ST, Lo CM, Ng IO, Liu CL, Lam CM, Wong J (2001) Improving survival results after resection of hepatocellular carcinoma: a prospective study of 377 patients over 10 years. Ann Surg 234(1):63–70 [PMID: 11420484]CrossRefPubMedGoogle Scholar
  4. 4.
    Xu KC, Niu LZ, He WB, Guo ZQ, Hu YZ, Zuo JS (2003) Percutaneous cryoablation in combination with ethanol injection for unresectable hepatocellular carcinoma. World J Gastroenterol 9(12):2686–2689 [PMID: 14669313]PubMedGoogle Scholar
  5. 5.
    Wang CP, Lu YY, Wang XZ, An LJ, Su SH, Zhou L, Chen Y, Jia HJ, Feng YY, Yang YP (2008) Percutaneous argon-helium cryoablation for primary hepatocellular carcinoma: report of 300 cases. Med J Chin PLA 133(12):1413–1417 (Beijing, China)Google Scholar
  6. 6.
    Helling TS (2000) Realistic expectations for cryoablation of liver tumors. J Hepatobiliary Pancreat Surg 7(5):510 [PMID: 11180879]CrossRefPubMedGoogle Scholar
  7. 7.
    Raut CP, Izzo F, Marra P, Ellis LM, Vauthey JN, Cremona F, Vallone P, Mastro A, Fornage BD, Curley SA (2005) Significant long-term survival after radiofrequency ablation of unresectable hepatocellular carcinoma in patients with cirrhosis. Ann Surg Oncol 12(8):616–628 [PMID: 15965731]CrossRefPubMedGoogle Scholar
  8. 8.
    Lencioni R, Cioni D, Crocetti L, Franchini C, Pina CD, Lera J, Bartolozzi C (2005) Early-stage hepatocellular carcinoma in patients with cirrhosis: long-term results of percutaneous image-guided radiofrequency ablation. Radiology 234(3):961–967 [PMID: 15665226]CrossRefPubMedGoogle Scholar
  9. 9.
    Kim YS, Rhim H, Cho OK, Koh BH, Kim Y (2006) Intrahepatic recurrence after percutaneous radiofrequency ablation of hepatocellular carcinoma: analysis of the pattern and risk factors. Eur J Radiol 59(3):432–441 [PMID: 16690240]CrossRefPubMedGoogle Scholar
  10. 10.
    Kudo M, Okanoue T (2007) Japan society of hepatology. Management of hepatocellular carcinoma in Japan: consensus-based clinical practice manual proposed by the Japan society of hepatology. Oncology 72(Suppl 1):2–15 [PMID: 18087177]CrossRefPubMedGoogle Scholar
  11. 11.
    Huang GW, Yang LY, Lu WQ (2005) Expression of hypoxiainducible factor 1alpha and vascular endothelial growth factor in hepatocellular carcinoma: impact on neovascularization and survival. World J Gastroenterol 11(11):1705–1708 [PMID: 15786555]PubMedGoogle Scholar
  12. 12.
    Poon RT, Ng IO, Lau C, Zhu LX, Yu WC, Lo CM, Fan ST, Wong J (2001) Serum vascular endothelial growth factor predicts venous invasion in hepatocellular carcinoma:a prospective study. Ann Surg 233(2):227–235 [PMID: 11176129]CrossRefPubMedGoogle Scholar
  13. 13.
    Kao JH, Chen PJ, Lai MY, Chen DS (2003) Basal core promoter mutations of hepatitis B virus increase the risk of hepatocellular carcinoma in hepatitis B carriers. Gastroenterology 124(2):327–334 [PMID: 12557138]CrossRefPubMedGoogle Scholar
  14. 14.
    Chen CJ, Yang HI, Su J, Jen CL, You SL, Lu SN, Huang GT, Iloeje UH (2006) Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level. JAMA 295(1):65–73 [PMID: 16391218]CrossRefPubMedGoogle Scholar
  15. 15.
    Yu MW, Yeh SH, Chen PJ, Liaw YF, Lin CL, Shih WL, Kao JH, Chen DS, Chen CJ (2005) Hepatitis B virus genotype and DNA level and hepatocellular carcinoma: a prospective study in men. J Natl Cancer Inst 97(4):265–272 [PMID: 15713961]PubMedGoogle Scholar
  16. 16.
    Bartholomeusz A, Locarnini S (2001) Hepatitis B virus mutants and fulminant hepatitis B: fitness plus phenotype. Hepatology 34(2):432–435 [PMID: 11481631]CrossRefPubMedGoogle Scholar
  17. 17.
    Liang TJ, Hasegawa K, Rimon N, Wands JR, Ben Porath E (1991) A hepatitis B virus mutant associated with an epidemic of fulminant hepatitis. N Engl J Med 324(24):1705–1709 [PMID: 2034247]PubMedCrossRefGoogle Scholar
  18. 18.
    Hunt CM, McGill JM, Allen MI, Condreay LD (2000) Clinical relevance of hepatitis B viral mutations. Hepatology 31(5):1037–1044 [PMID: 10796877]CrossRefPubMedGoogle Scholar
  19. 19.
    Lin CL, Liao LY, Wang CS, Chen PJ, Lai MY, Chen DS, Kao JH (2005) Basal core-promoter mutant of hepatitis B virus and progression of liver disease in hepatitis B e antigen-negative chronic hepatitis B. Liver Int 25(3):564–570 [PMID: 15910494]CrossRefPubMedGoogle Scholar
  20. 20.
    Liu CJ, Chen BF, Chen PJ, Lai MY, Huang WL, Kao JH, Chen DS (2006) Role of hepatitis B viral load and basalcore promoter mutation in hepatocellular carcinoma in hepatitis B carriers. J Infect Dis 193(9):1258–1265 [PMID: 16586363]CrossRefPubMedGoogle Scholar
  21. 21.
    Wang ZL, Liang P, Dong BW, Yu XL, De Yu J (2008) Prognostic factors and recurrence of small hepatocellular carcinoma after hepatic resection or microwave ablation: a retrospective study. J Gastrointest Surg 12(2):327–337 [PMID: 17943391]CrossRefPubMedGoogle Scholar
  22. 22.
    Poon RT, Lau C, Pang R, Ng KK, Yuen J, Fan ST (2007) High serum vascular endothelial growth factor levels predict poor prognosis after radiofrequency ablation of hepatocellular carcinoma: importance of tumor biomarker in ablative therapies. Ann Surg Oncol 14(6):1835–1845 [PMID: 17406950]CrossRefPubMedGoogle Scholar
  23. 23.
    Poon RT, Ho JW, Tong CS, Lau C, Ng IO, Fan ST (2004) Prognostic significance of serum vascular endothelial growth factor and endostatin in patients with hepatocellular carcinoma. Br J Surg 91(10):1354–1360 [PMID: 15376182]CrossRefPubMedGoogle Scholar
  24. 24.
    Chao Y, Li CP, Chau GY, Chen CP, King KL, Lui WY, Yen SH, Chang FY, Chan WK, Lee SD (2003) Prognostic significance of vascular endothelial growth factor, basic fibroblast growth factor, and angiogenin in patients with resectable hepatocellular carcinoma after surgery. Ann Surg Oncol 10(4):355–362 [PMID: 12734082]CrossRefPubMedGoogle Scholar
  25. 25.
    Li X, Feng GS, Zheng CS, Zhuo CK, Liu X (2004) Expression of plasma vascular endothelial growth factor in patients with hepatocellular carcinoma and effect of transcatheter arterial chemoembolization therapy on plasma vascular endothelial growth factor level. World J Gastroenterol 10(19):2878–2882 [PMID: 15334691]PubMedGoogle Scholar
  26. 26.
    Salizzoni M, Romagnoli R, Lupo F, David E, Mirabella S, Cerutti E, Ottobrelli A (2003) Microscopic vascular invasion detected by anti-CD34 immunohistochemistry as a predictor of recurrence of hepatocellular carcinoma after liver transplantation. Transplantation 76(5):844–848 [PMID: 14501865]CrossRefPubMedGoogle Scholar
  27. 27.
    Jonas S, Bechstein WO, Steinmüller T, Herrmann M, Radke C, Berg T, Settmacher U, Neuhaus P (2001) Vascular invasion and histopathologic grading determine outcome after liver transplantation for hepatocellular carcinoma in cirrhosis. Hepatology 33(5):1080–1086 [PMID: 11343235]CrossRefPubMedGoogle Scholar
  28. 28.
    Poon RT, Lau CP, Cheung ST, Yu WC, Fan ST (2003) Quantitative correlation of serum levels and tumor expression of vascular endothelial growth factor in patients with hepatocellular carcinoma. Cancer Res 63(12):3121–3126 [PMID: 12810638]PubMedGoogle Scholar
  29. 29.
    Jinno K, Tanimizu M, Hyodo I, Nishikawa Y, Hosokawa Y, Doi T, Endo H, Yamashita T, Okada Y (1998) Circulating vascular endothelial growth factor (VEGF) is a possible tumor marker for metastasis in human hepatocellular carcinoma. J Gastroenterol 33(3):376–382 [PMID: 9658317]CrossRefPubMedGoogle Scholar
  30. 30.
    Poon RT, Fan ST, Wong J (2001) Clinical implications of circulating angiogenic factors in cancer patients. J Clin Oncol 19(4):1207–1225 [PMID: 11181687]PubMedGoogle Scholar
  31. 31.
    George ML, Eccles SA, Tutton MG, Abulafi AM, Swift RI (2000) Correlation of plasma and serum vascular endothelial growth factor levels with platelet count in colorectal cancer: clinical evidence of platelet scavenging? Clin Cancer Res 6(8):3147–3152 [PMID: 10955796]PubMedGoogle Scholar
  32. 32.
    Li B, Sharpe EE, Maupin AB, Teleron AA, Pyle AL, Carmeliet P, Young PP (2006) VEGF and PlGF promote adult vasculogenesis by enhancing EPC recruitment and vessel formation at the site of tumor neovascularization. FASEB J 20(9):1495–1497 [ PMID: 16754748]CrossRefPubMedGoogle Scholar
  33. 33.
    Moore MA, Hattori K, Heissig B, Shieh JH, Dias S, Crystal RG, Rafii S (2001) Mobilization of endothelial and hematopoietic stem and progenitor cells by adenovector-mediated elevation of serum levels of SDF-1, VEGF, and angiopoietin-1. Ann N Y Acad Sci 938:36–45 [PMID: 11458524]PubMedGoogle Scholar
  34. 34.
    Ho JW, Pang RW, Lau C, Sun CK, Yu WC, Fan ST, Poon RT (2006) Significance of circulating endothelial progenitor cells in hepatocellular carcinoma. Hepatology 44(4):836–843 [PMID: 17006919]CrossRefPubMedGoogle Scholar
  35. 35.
    Machi J, Bueno RS, Wong LL (2005) Long-term follow-up outcome of patients undergoing radiofrequency ablation for unresectable hepatocellular carcinoma. World J Surg 29(11):1364–1373 [PMID: 16240062]CrossRefPubMedGoogle Scholar
  36. 36.
    Xu HX, Lu MD, Xie XY, Yin XY, Kuang M, Chen JW, Xu ZF, Liu GJ (2005) Prognostic factors for long term outcome after percutaneous thermal ablation for hepatocellular carcinoma: a survival analysis of 137 consecutive patients. Clin Radiol 60(9):1018–1025 [PMID: 16124984]CrossRefPubMedGoogle Scholar
  37. 37.
    Chung IK, Park MJ, Kwon KT, Park YD, Chung YJ, Jeon SW, Lee MK, Seo HE, Lee YD, Cho CM, Tak WY, Kweon YO, Kim SK, Choi YH (2005) The factors related to the prognosis of solitary hepatocellular carcinoma after radiofrequency ablation. Korean J Hepatol 11(4):371–380 [PMID: 16380666]PubMedGoogle Scholar
  38. 38.
    Lin CL, Chen JD, Liu CJ, Lee PH, Chen PJ, Lai MY, Kao JH, Chen DS (2007) Clinicopathological differences between hepatitis B viral genotype B- and C-related resectable hepatocellular carcinoma. J Viral Hepat 14(1):64–69 [PMID: 17212646]CrossRefPubMedGoogle Scholar
  39. 39.
    Yang HI, Yeh SH, Chen PJ, Iloeje UH, Jen CL, Su J, Wang LY, Lu SN, You SL, Chen DS, Liaw YF, Chen CJ (2008) REVEAL-HBV study group. Associations between hepatitis B virus genotype and mutants and the risk of hepatocellular carcinoma. J Natl Cancer Inst 100(16):1134–1143 [PMID: 18695135]CrossRefPubMedGoogle Scholar
  40. 40.
    Huang Y, Wang Z, An S, Zhou B, Zhou Y, Chan HL, Hou J (2008) Role of hepatitis B virus genotypes and quantitative HBV DNA in metastasis and recurrence of hepatocellular carcinoma. J Med Virol 80(4):591–597 [PMID: 18297705 ]CrossRefPubMedGoogle Scholar
  41. 41.
    Dong Q, Chan HL, Liu Z, Chan DP, Zhang B, Chen Y, Kung HF, Sung JJ, He ML (2008) A1762T/G1764A mutations of hepatitis B virus, associated with the increased risk of hepatocellular carcinoma, reduce basal core promoter activities. Biochem Biophys Res Commun 374(4):773–776 [PMID: 18675784]CrossRefPubMedGoogle Scholar
  42. 42.
    Yuan J, Zhou B, Tanaka Y, Kurbanov F, Orito E, Gong Z, Xu L, Lu J, Jiang X, Lai W, Mizokami M (2007) Hepatitis B virus (HBV) genotypes/subgenotypes in China: mutations in core promoter and precore/core and their clinical implications. J Clin Virol 39(2):87–93 [PMID: 17451999]CrossRefPubMedGoogle Scholar
  43. 43.
    Chou YC, Yu MW, Wu CF, Yang SY, Lin CL, Liu CJ, Shih WL, Chen PJ, Liaw YF, Chen CJ (2008) Temporal relationship between hepatitis B virus enhancer II/basal core promoter sequence variation and risk of hepatocellular carcinoma. Gut 57(1):91–97 [PMID: 17502344]CrossRefPubMedGoogle Scholar
  44. 44.
    Wu CF, Yu MW, Lin CL, Liu CJ, Shih WL, Tsai KS, Chen CJ (2008) Long-term tracking of hepatitis B viral load and the relationship with risk for hepatocellular carcinoma in men. Carcinogenesis 29(1):106–112 [PMID: 17999990]CrossRefPubMedGoogle Scholar
  45. 45.
    Shinkai N, Tanaka Y, Ito K, Mukaide M, Hasegawa I, Asahina Y, Izumi N, Yatsuhashi H, Orito E, Joh T, Mizokami M (2007) Influence of hepatitis B virus X and core promoter mutations on hepatocellular carcinoma among patients infected with subgenotype C2. J Clin Microbiol 45(10):3191–3197 [PMID: 17652471]CrossRefPubMedGoogle Scholar
  46. 46.
    Sung JJ, Tsui SK, Tse CH, Ng EY, Leung KS, Lee KH, Mok TS, Bartholomeusz A, Au TC, Tsoi KK, Locarnini S, Chan HL (2008) Genotype-specific genomic markers associated with primary hepatomas, based on complete genomic sequencing of hepatitis B virus. J Virol 82(7):3604–3611 [PMID: 18216102]CrossRefPubMedGoogle Scholar
  47. 47.
    Sakon M, Umeshita K, Nagano H, Eguchi H, Kishimoto S, Miyamoto A, Ohshima S, Dono K, Nakamori S, Gotoh M, Monden M (2000) Clinical significance of hepatic resection in hepatocellular carcinoma analysis by disease-free survival curves. Arch Surg 135(12):1456–1459 [PMID: 11115352]CrossRefPubMedGoogle Scholar
  48. 48.
    Sumi H, Yokosuka O, Seki N, Arai M, Imazeki F, Kurihara T, Kanda T, Fukai K, Kato M, Saisho H (2003) Influence of hepatitis B virus genotypes on the progression of chronic type B liver disease. Hepatology 37(1):19–26 [PMID: 12500184]CrossRefPubMedGoogle Scholar
  49. 49.
    Yuen MF, Tanaka Y, Mizokami M, Yuen JC, Wong DK, Yuan HJ, Sum SM, Chan AO, Wong BC, Lai CL (2004) Role of hepatitis B virus genotypes Ba and C, core promoter and precore mutations on hepatocellular carcinoma: a case control study. Carcinogenesis 25(9):1593–1598 [PMID: 15090469]CrossRefPubMedGoogle Scholar
  50. 50.
    Zhu AX, Blaszkowsky LS, Ryan DP, Clark JW, Muzikansky A, Horgan K, Sheehan S, Hale KE, Enzinger PC, Bhargava P, Stuart K (2006) Phase II study of gemcitabine and oxaliplatin in combination with bevacizumab in patients with advanced hepatocellular carcinoma. J Clin Oncol 24(12):1898–1903 [PMID: 16622265]CrossRefPubMedGoogle Scholar

Copyright information

© Springer 2009

Authors and Affiliations

  • Chunping Wang
    • 1
  • Yinying Lu
    • 1
  • Yan Chen
    • 1
  • Yongyi Feng
    • 1
  • Linjing An
    • 1
  • Xinzhen Wang
    • 1
  • Shuhui Su
    • 1
  • Wenlin Bai
    • 1
  • Lin Zhou
    • 1
  • Yongping Yang
    • 1
  • Dongping Xu
    • 2
  1. 1.Center of Therapeutic Research for Hepatocellular CarcinomaBeijingChina
  2. 2.Infectious Disease Institute of PLABeijingChina

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