Next-Generation Sequencing-Based Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma Predicts Hepatocellular Carcinoma Recurrence
Abstract
:1. Introduction
2. Materials and Methods
2.1. Human Sample Collection
2.2. NGS-Based Platform for Pre-S Genotyping
2.3. Statistical Analysis
3. Results
3.1. Clinicopathological Profiles of the HBV-Related HCC Patients
3.2. Pre-S Genotyping and Patient Grouping by the NGS-Based Platform
3.3. Patients with Either Deletions Spanning the Pre-S2 Gene Segment or High Percentage of Pre-S2 Plus Pre-S1 + Pre-S2 Deletions as a High-Risk Population for HCC Recurrence after Surgical Resection
3.4. Combination of Deletions Spanning the Pre-S2 Gene Segment and the Percentage of Pre-S2 Plus Pre-S1 + Pre-S2 Deletions as an Independent Prognostic Factor for HCC Recurrence after Surgical Resection
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Llovet, J.M.; Zucman-Rossi, J.; Pikarsky, E.; Sangro, B.; Schwartz, M.; Sherman, M.; Gores, G. Hepatocellular carcinoma. Nat. Rev. Dis. Prim. 2016, 2, 16019. [Google Scholar] [CrossRef] [PubMed]
- Venook, A.P.; Papandreou, C.; Furuse, J.; De Guevara, L.L. The Incidence and Epidemiology of Hepatocellular Carcinoma: A Global and Regional Perspective. Oncologist 2010, 15, 5–13. [Google Scholar] [CrossRef] [PubMed]
- Cheng, K.-C.; Lin, W.-Y.; Liu, C.-S.; Lin, C.-C.; Lai, H.-C.; Lai, S.-W. Association of different types of liver disease with demographic and clinical factors. Biomedicine 2016, 6, 16. [Google Scholar] [CrossRef] [PubMed]
- Llovet, J.M.; Bruix, J. Novel advancements in the management of hepatocellular carcinoma in 2008. J. Hepatol. 2008, 48, S20–S37. [Google Scholar] [CrossRef]
- Wall, W.J.; Marotta, P.J. Surgery and transplantation for hepatocellular cancer. Liver Transplant. 2000, 6, s16–s22. [Google Scholar] [CrossRef] [PubMed]
- Alsowmely, A.M.; Hodgson, H.J. Non-surgical treatment of hepatocellular carcinoma. Aliment. Pharmacol. Ther. 2002, 16, 1–15. [Google Scholar] [CrossRef]
- Llovet, J.M.; Ricci, S.; Mazzaferro, V.M.; Hilgard, P.; Gane, E.; Blanc, J.-F.; De Oliveira, A.C.; Santoro, A.; Raoul, J.L.; Forner, A.; et al. Sorafenib in Advanced Hepatocellular Carcinoma. N. Engl. J. Med. 2008, 359, 378–390. [Google Scholar] [CrossRef]
- Marín-Hargreaves, G.; Azoulay, D.; Bismuth, H. Hepatocellular carcinoma: Surgical indications and results. Crit. Rev. Oncol. 2003, 47, 13–27. [Google Scholar] [CrossRef]
- Poon, R.T.-P.; Fan, S.-T.; Wong, J. Risk Factors, Prevention, and Management of Postoperative Recurrence After Resection of Hepatocellular Carcinoma. Ann. Surg. 2000, 232, 10–24. [Google Scholar] [CrossRef]
- Poon, R.T.-P.; Fan, S.-T.; Lo, C.-M.; Liu, C.-L.; Wong, J. Intrahepatic Recurrence After Curative Resection of Hepatocellular Carcinoma. Ann. Surg. 1999, 229, 216–222. [Google Scholar] [CrossRef]
- Beasley, R.; Hwang, L.-Y. Hepatocellular Carcinoma and Hepatitis B Virus. Semin. Liver Dis. 1984, 4, 113–121. [Google Scholar] [CrossRef] [PubMed]
- Bosetti, C.; Turati, F.; La Vecchia, C. Hepatocellular carcinoma epidemiology. Best Pr. Res. Clin. Gastroenterol. 2014, 28, 753–770. [Google Scholar] [CrossRef] [PubMed]
- Su, I.-J.; Wang, H.-C.; Wu, H.-J.; Huang, W.-Y. Ground glass hepatocytes contain pre-S mutants and represent preneoplastic lesions in chronic hepatitis B virus infection. J. Gastroenterol. Hepatol. 2008, 23, 1169–1174. [Google Scholar] [CrossRef] [PubMed]
- Fan, Y.; Lu, C.; Chang, Y.; Chang, T.; Lin, P.; Lei, H.; Su, I. Identification of a pre-S2 mutant in hepatocytes expressing a novel marginal pattern of surface antigen in advanced diseases of chronic hepatitis B virus infection. J. Gastroenterol. Hepatol. 2000, 15, 519–528. [Google Scholar] [CrossRef]
- Wang, H.-C.; Wu, H.-J.; Chen, C.-F.; Fausto, N.; Lei, H.-Y.; Su, I.-J. Different Types of Ground Glass Hepatocytes in Chronic Hepatitis B Virus Infection Contain Specific Pre-S Mutants that May Induce Endoplasmic Reticulum Stress. Am. J. Pathol. 2003, 163, 2441–2449. [Google Scholar] [CrossRef] [Green Version]
- Teng, C.-F.; Wu, H.-J.; Shyu, W.-C.; Jeng, L.-B.; Su, I.-J. Pre-S2 Mutant-Induced Mammalian Target of Rapamycin Signal Pathways as Potential Therapeutic Targets for Hepatitis B Virus-Associated Hepatocellular Carcinoma. Cell Transplant. 2017, 26, 429–438. [Google Scholar] [CrossRef] [Green Version]
- Teng, Y.-C.; Neo, J.C.; Wu, J.-C.; Chen, Y.-F.; Kao, C.-H.; Tsai, T.-F. Expression of a hepatitis B virus pre-S2 deletion mutant in the liver results in hepatomegaly and hepatocellular carcinoma in mice. J. Pathol. 2017, 241, 463–474. [Google Scholar] [CrossRef]
- Shen, F.-C.; Su, I.-J.; Wu, H.-J.; Hsieh, Y.-H.; Yao, W.-J.; Young, K.-C.; Chang, T.C.; Hsieh, H.-C.; Tsai, H.-N.; Huang, W.-H. A pre-S gene chip to detect pre-S deletions in hepatitis B virus large surface antigen as a predictive marker for hepatoma risk in chronic hepatitis B virus carriers. J. Biomed. Sci. 2009, 16, 84. [Google Scholar] [CrossRef] [Green Version]
- Chen, C.; Hung, C.; Chen, W.J.; Hu, T.; Wang, J.; Wang, J.; Lu, S.-N.; Changchien, C. Pre-S Deletion and Complex Mutations of Hepatitis B Virus Related to Advanced Liver Disease in HBeAg-Negative Patients. Gastroenterology 2007, 133, 1466–1474. [Google Scholar] [CrossRef]
- Sinn, D.H.; Choi, M.S.; Gwak, G.-Y.; Paik, Y.H.; Lee, J.H.; Koh, K.C.; Paik, S.W.; Yoo, B.C. Pre-S Mutation Is a Significant Risk Factor for Hepatocellular Carcinoma Development: A Long-Term Retrospective Cohort Study. Dig. Dis. Sci. 2012, 58, 751–758. [Google Scholar] [CrossRef]
- Fan, Y.; Lu, C.-C.; Chen, W.; Yao, W.-J.; Wang, H.; Chang, T.; Lei, H.-Y.; Shiau, A.-L.; Su, I. Prevalence and significance of hepatitis B virus (HBV) pre-S mutants in serum and liver at different replicative stages of chronic HBV infection. Hepatology 2001, 33, 277–286. [Google Scholar] [CrossRef] [PubMed]
- Tsai, H.-W.; Lin, Y.-J.; Lin, P.-W.; Wu, H.-J.; Hsu, K.-H.; Yen, C.-J.; Chan, S.-H.; Huang, W.-H.; Su, I.-J. A clustered ground-glass hepatocyte pattern represents a new prognostic marker for the recurrence of hepatocellular carcinoma after surgery. Cancer 2011, 117, 2951–2960. [Google Scholar] [CrossRef] [PubMed]
- Tsai, H.-W.; Lin, Y.-J.; Wu, H.-C.; Chang, T.-T.; Wu, I.-C.; Cheng, P.-N.; Yen, C.-J.; Chan, S.-H.; Huang, W.; Su, I.-J. Resistance of ground glass hepatocytes to oral antivirals in chronic hepatitis B patients and implication for the development of hepatocellular carcinoma. Oncotarget 2016, 7, 27724–27734. [Google Scholar] [CrossRef] [PubMed]
- Yen, C.-J.; Ai, Y.-L.; Tsai, H.-W.; Chan, S.-H.; Yen, C.-S.; Cheng, K.-H.; Lee, Y.-P.; Kao, C.-W.; Wang, Y.-C.; Chen, Y.-L.; et al. Hepatitis B virus surface gene pre-S2 mutant as a high-risk serum marker for hepatoma recurrence after curative hepatic resection. Hepatology 2018, 68, 815–826. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Teng, C.-F.; Huang, H.-Y.; Li, T.-C.; Shyu, W.-C.; Wu, H.-J.; Lin, C.-Y.; Su, I.-J.; Jeng, L.-B. A Next-Generation Sequencing-Based Platform for Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma of Hepatocellular Carcinoma Patients. Sci. Rep. 2018, 8, 14816. [Google Scholar] [CrossRef]
- Teng, C.-F.; Tsai, H.-W.; Li, T.-C.; Wang, T.; Wang, J.; Shyu, W.-C.; Wu, H.-C.; Su, I.-J.; Jeng, L.-B. Detection of hepatitis B virus pre-S mutants in plasma by a next-generation sequencing-based platform determines their patterns in liver tissues. PLoS ONE 2020, 15, e0234773. [Google Scholar] [CrossRef]
- Lee, S.C.; Tan, H.T.; Chung, M.C.M. Prognostic biomarkers for prediction of recurrence of hepatocellular carcinoma: Current status and future prospects. World J. Gastroenterol. 2014, 20, 3112–3124. [Google Scholar] [CrossRef]
- Zulehner, G.; Mikula, M.; Schneller, R.; Van Zijl, F.; Huber, H.; Sieghart, W.; Grasl-Kraupp, B.; Waldhör, T.; Peck-Radosavljevic, M.; Beug, H.; et al. Nuclear β-Catenin Induces an Early Liver Progenitor Phenotype in Hepatocellular Carcinoma and Promotes Tumor Recurrence. Am. J. Pathol. 2010, 176, 472–481. [Google Scholar] [CrossRef]
- Yin, X.; Li, Y.-W.; Jin, J.-J.; Zhou, Y.; Ren, Z.-G.; Qiu, S.-J.; Zhang, B.-H. The clinical and prognostic implications of pluripotent stem cell gene expression in hepatocellular carcinoma. Oncol. Lett. 2013, 5, 1155–1162. [Google Scholar] [CrossRef] [Green Version]
- Hu, T.; Wang, C.-C.; Huang, C.-C.; Chen, C.-L.; Hung, C.-H.; Chen, C.-H.; Wang, J.-H.; Lu, S.-N.; Lee, C.-M.; Changchien, C.-S.; et al. Down-regulation of tumor suppressor gene PTEN, overexpression of p53, plus high proliferating cell nuclear antigen index predict poor patient outcome of hepatocellular carcinoma after resection. Oncol. Rep. 2007, 18, 1417–1426. [Google Scholar] [CrossRef]
- Yu, S.J.; Kim, Y.J. Hepatitis B viral load affects prognosis of hepatocellular carcinoma. World J. Gastroenterol. 2014, 20, 12039–12044. [Google Scholar] [CrossRef] [PubMed]
- Yang, T.; Lu, J.-H.; Zhai, J.; Lin, C.; Yang, G.-S.; Zhao, R.-H.; Shen, F.; Wu, M.-C. High viral load is associated with poor overall and recurrence-free survival of hepatitis B virus-related hepatocellular carcinoma after curative resection: A prospective cohort study. Eur. J. Surg. Oncol. (EJSO) 2012, 38, 683–691. [Google Scholar] [CrossRef] [PubMed]
- Sohn, W.; Paik, Y.-H.; Kim, J.M.; Kwon, A.C.H.D.; Joh, J.-W.; Cho, J.Y.; Gwak, G.-Y.; Choi, M.S.; Lee, J.H.; Koh, K.C.; et al. HBV DNA and HBsAg Levels as Risk Predictors of Early and Late Recurrence after Curative Resection of HBV-related Hepatocellular Carcinoma. Ann. Surg. Oncol. 2014, 21, 2429–2435. [Google Scholar] [CrossRef] [PubMed]
- Wong, J.S.-W.; Wong, G.L.-H.; Tsoi, K.K.-F.; Wong, V.W.-S.; Cheung, S.Y.-S.; Chong, C.-N.; Lee, K.-F.; Lai, P.B.-S.; Chan, H.L.Y. Meta-analysis: The efficacy of anti-viral therapy in prevention of recurrence after curative treatment of chronic hepatitis B-related hepatocellular carcinoma. Aliment. Pharmacol. Ther. 2011, 33, 1104–1112. [Google Scholar] [CrossRef]
- Miao, R.-Y.; Zhao, H.-T.; Yang, H.-Y.; Mao, Y.-L.; Lü, X.; Zhao, Y.; Liu, C.-N.; Zhong, S.-X.; Sang, X.-T.; Huang, J.-F. Postoperative adjuvant antiviral therapy for hepatitis B/C virus-related hepatocellular carcinoma: A meta-analysis. World J. Gastroenterol. 2010, 16, 2931–2942. [Google Scholar] [CrossRef]
- Xu, M.; Zhou, Z.; Xu, R.; Lin, N.; Lin, N.; Zhong, Y. Antiviral therapy predicts the outcomes following resection of hepatocellular carcinoma in patients negative for HBV DNA: A propensity score matching analysis. World J. Surg. Oncol. 2019, 17, 45. [Google Scholar] [CrossRef]
- Morikawa, K.; Suda, G.; Sakamoto, N. Viral life cycle of hepatitis B virus: Host factors and druggable targets. Hepatol. Res. 2016, 46, 871–877. [Google Scholar] [CrossRef]
- Jilbert, A.R.; Reaiche, G.Y.; Clouston, A.; Test, S. Hepatitis B Virus. eLS 2011, 26, 386–387. [Google Scholar] [CrossRef]
- Hu, J.; Protzer, U.; Siddiqui, A.; Protzer, U. Revisiting Hepatitis B Virus: Challenges of Curative Therapies. J. Virol. 2019, 93. [Google Scholar] [CrossRef] [Green Version]
- Levrero, M.; Zucman-Rossi, J. Mechanisms of HBV-induced hepatocellular carcinoma. J. Hepatol. 2016, 64, S84–S101. [Google Scholar] [CrossRef]
- Tu, T.; Budzinska, M.A.; Shackel, N.A.; Urban, S. HBV DNA Integration: Molecular Mechanisms and Clinical Implications. Viruses 2017, 9, 75. [Google Scholar] [CrossRef] [PubMed]
- Wang, M.; Xi, N.; Ning, Q. Virus-induced hepatocellular carcinoma with special emphasis on HBV. Hepatol. Int. 2017, 11, 171–180. [Google Scholar] [CrossRef] [PubMed]
- Sung, W.-K.; Zheng, H.; Li, S.; Chen, R.; Liu, X.; Li, Y.; Lee, N.P.; Lee, W.H.; Ariyaratne, P.N.; Tennakoon, C.; et al. Genome-wide survey of recurrent HBV integration in hepatocellular carcinoma. Nat. Genet. 2012, 44, 765–769. [Google Scholar] [CrossRef] [PubMed]
- Tarocchi, M.; Polvani, S.; Marroncini, G.; Galli, A. Molecular mechanism of hepatitis B virus-induced hepatocarcinogenesis. World J. Gastroenterol. 2014, 20, 11630–11640. [Google Scholar] [CrossRef] [PubMed]
- Jia, J.; Liang, X.; Chen, S.; Wang, H.; Li, H.; Fang, M.; Bai, X.; Wang, Z.; Wang, M.; Zhu, S.; et al. Next-generation sequencing revealed divergence in deletions of the preS region in the HBV genome between different HBV-related liver diseases. J. Gen. Virol. 2017, 98, 2748–2758. [Google Scholar] [CrossRef] [PubMed]
Characteristics | No. of Patients | Median (Range) |
---|---|---|
Age (years) | 75 | 53 (26–78) |
>50 | 48 | 60 (51–78) |
≤50 | 27 | 43 (26–50) |
Gender (men/women) | 68/7 | |
Smoking (yes/no) | 31/44 | |
Alcohol (yes/no) | 29/46 | |
HBsAg (positive/negative/NA) | 65/0/10 | |
HBeAg (positive/negative/NA) | 9/62/4 | |
HBV genotype (B/C) | 60/15 | |
HBV DNA (IU/mL) (20–1.7 × 108/<20) a | 74/1 | 2.1 × 104 (21.5–1.5 × 108) c |
>1 × 104 | 42 | 4.3 × 105 (1.2 × 104–1.5 × 108) |
≤1 × 104 | 32 | 8.4 × 102 (21.5–9.3 × 103) |
Albumin (g/dL) | 75 | 3.7 (1.2–4.9) |
>3.8 | 30 | 4.2 (3.9–4.9) |
≤3.8 | 45 | 3.3 (1.2–3.8) |
AST (U/L) | 75 | 60 (14–1052) |
>34 | 61 | 79 (35–1052) |
≤34 | 14 | 27 (14–34) |
ALT (U/L) | 75 | 55 (13–1338) |
>40 | 50 | 96.5 (41–1338) |
≤40 | 25 | 31 (13–40) |
AFP (ng/mL) (≤54,000/>54,000) b | 71/4 | 26.7 (1.8–36,600.0) d |
>400 | 28 | 1920 (461.7–36,600.0) |
≤400 | 47 | 13.8 (1.8–271.0) |
Tumor size (cm) | 75 | 4.5 (1.1–19.5) |
>5 | 37 | 10.0 (5.5–19.5) |
≤5 | 38 | 2.4 (1.1–4.5) |
Tumor encapsulation (yes/no/NA) | 42/20/13 | |
Lymph node involvement (yes/no) | 8/67 | |
Portal vein thrombosis (yes/no) | 5/70 | |
Vascular invasion (yes/no) | 27/48 | |
Distant metastasis (yes/no) | 8/67 | |
Steatosis grade (0/1/2/3/NA) | 14/10/1/0/50 | |
Metavir inflammation score (0/1/2/3/NA) | 4/35/5/0/31 | |
Ishak fibrosis score (0/1/2/3/4/5/6/NA) | 5/13/12/8/3/4/11/19 | |
Child-Pugh cirrhosis score (A/B/C) | 57/16/2 | |
CLIP score (0/1/2/3/4/5/6) | 33/23/10/8/1/0/0 | |
Tumor differentiation grade (1/2/3/4) | 2/36/36/1 | |
BCLC stage (A/B/C/D) | 38/29/7/1 | |
AJCC TNM stage (I/II/IIIA/IIIB/IIIC/IVA/IVB) | 40/20/7/5/3/0/0 | |
Antiviral therapy after surgery (yes/no) | 40/35 | |
HCC recurrence after surgery (month) (yes/no) | 52/23 | 11.2 (1.5–72.3) e |
Survival after surgery (month) (dead/alive) | 16/59 | 26.9 (6.8–161.1) f |
Pre-S Del Type of Patients a | No. of Patients (%) b |
---|---|
Total patients | 75 (100) |
Patients without del spanning pre-S gene segments | 29 (39) |
Patients with del spanning pre-S gene segments | 46 (61) |
Patients with only pre-S1 del | 15 (20) |
Patients with only pre-S2 del | 6 (8) |
Patients with only pre-S1 + pre-S2 del | 0 (0) |
Patients with both pre-S1 and pre-S2 del | 7 (9) |
Patients with both pre-S1 and pre-S1 + pre-S2 del | 4 (5) |
Patients with both pre-S2 and pre-S1 + pre-S2 del | 2 (3) |
Patients with all three types of pre-S del | 12 (16) |
Patients with del spanning the pre-S1 gene segment | 40 (53) |
Patients with del spanning the pre-S2 gene segment | 31 (41) |
Pre-S Del Percentage of Patients | No. of Patients (%) |
Pre-S1 del percentage c | 75 (100) |
I (≤1.895) d | 19 (25) |
II (>1.895) | 19 (25) |
III (>5.629) | 18 (24) |
IV (>28.270) | 19 (25) |
Pre-S2 del percentage | 75 (100) |
I (≤0.643) | 19 (25) |
II (>0.643) | 19 (25) |
III (>1.824) | 18 (24) |
IV (>13.666) | 19 (25) |
Pre-S1 + pre-S2 del percentage | 75 (100) |
I (≤0.015) | 19 (25) |
II (>0.015) | 18 (24) |
III (>0.442) | 19 (25) |
IV (>2.975) | 19 (25) |
Pre-S1 plus pre-S2 del percentage | 75 (100) |
I (≤2.725) | 18 (24) |
II (>2.725) | 19 (25) |
III (>18.539) | 19 (25) |
IV (>49.938) | 19 (25) |
Pre-S1 plus pre-S1 + pre-S2 del percentage | 75 (100) |
I (≤2.043) | 18 (24) |
II (>2.043) | 19 (25) |
III (>6.332) | 20 (27) |
IV (>44.201) | 18 (24) |
Pre-S2 plus pre-S1 + pre-S2 del percentage | 75 (100) |
I (≤0.689) | 18 (24) |
II (>0.689) | 20 (27) |
III (>2.109) | 19 (25) |
IV (>24.995) | 18 (24) |
All three types of pre-S del percentage | 75 (100) |
I (≤2.828) | 18 (24) |
II (>2.828) | 20 (27) |
III (>21.666) | 18 (24) |
IV (>58.035) | 19 (25) |
Characteristics | Univariate Analysis | Multivariate Analysis | ||||
---|---|---|---|---|---|---|
HR | 95% CI | p Value | HR | 95% CI | p Value | |
Age (years) (>50 vs. ≤50) | 0.951 | 0.532–1.700 | 0.8666 | |||
Gender (men vs. women) | 1.043 | 0.414–2.627 | 0.9284 | |||
Smoking (yes vs. no) | 0.886 | 0.503–1.560 | 0.6750 | |||
Alcohol (yes vs. no) | 0.884 | 0.494–1.580 | 0.6773 | |||
HBsAg (positive vs. negative) a | ||||||
HBeAg (positive vs. negative) b | 1.234 | 0.523–2.910 | 0.6307 | |||
HBV genotype (B vs. C) | 0.583 | 0.304–1.117 | 0.1040 | |||
HBV DNA (IU/mL) (>1 × 104 vs. ≤1 × 104) c | 1.645 | 0.934–2.895 | 0.0846 | |||
Albumin (g/dL) (>3.8 vs. ≤3.8) | 0.551 | 0.288–1.092 | 0.0585 | |||
AST (U/L) (>34 vs. ≤34) | 0.865 | 0.444–1.684 | 0.6691 | |||
ALT (U/L) (>40 vs. ≤40) | 0.797 | 0.456–1.394 | 0.4267 | |||
AFP (ng/mL) (>400 vs. ≤400) | 1.305 | 0.745–2.285 | 0.3524 | |||
Tumor size (cm) (>5 vs. ≤5) | 1.490 | 0.863–2.572 | 0.1525 | |||
Tumor encapsulation (yes vs. no) d | 0.901 | 0.474–1.713 | 0.7508 | |||
Lymph node involvement (yes vs. no) | 0.333 | 0.104–1.071 | 0.0652 | |||
Portal vein thrombosis (yes vs. no) | 1.668 | 0.600–4.633 | 0.3264 | |||
Vascular invasion (yes vs. no) | 1.677 | 0.962–2.924 | 0.0681 | |||
Distant metastasis (yes vs. no) | 2.259 | 0.999–5.101 | 0.0502 | |||
Steatosis grade (2/3 vs. 0/1) e | 3.473 | 0.418–28.879 | 0.2493 | |||
Metavir inflammation score (2/3 vs. 0/1) f | 0.731 | 0.256–2.088 | 0.5583 | |||
Ishak fibrosis score (4/5/6 vs. 0/1/2/3) g | 1.261 | 0.670–2.373 | 0.4714 | |||
Child-Pugh cirrhosis score (B/C vs. A) | 2.189 | 1.195–4.013 | 0.0112 * | 2.182 | 1.142–4.171 | 0.0182 * |
CLIP score (4/5/6 vs. 0/1/2/3) | 2.426 | 0.328–17.911 | 0.3850 | |||
Tumor differentiation (3/4 vs. 1/2) | 1.246 | 0.722–2.150 | 0.4288 | |||
BCLC stage (C/D vs. A/B) | 1.927 | 0.867–4.284 | 0.1077 | |||
AJCC TNM stage (IIIA/IIIB/IIIC/IVA/IVB vs. I/II) | 4.048 | 2.123–7.719 | <0.0001 *** | 3.667 | 1.853–7.258 | 0.0002 *** |
Antiviral therapy after surgery (yes vs. no) | 1.176 | 0.674–2.051 | 0.5684 | |||
Del spanning pre-S gene segments (yes vs. no) | 1.315 | 0.742–2.332 | 0.3477 | |||
Only pre-S1 del (yes vs. no) | 0.590 | 0.277–1.257 | 0.1715 | |||
Only pre-S2 del (yes vs. no) | 1.421 | 0.564–3.580 | 0.4557 | |||
Only pre-S1 + pre-S2 del (yes vs. no) h | ||||||
Both pre-S1 and pre-S2 del (yes vs. no) | 1.260 | 0.453–3.505 | 0.6573 | |||
Both pre-S1 and pre-S1 + pre-S2 del (yes vs. no) | 2.460 | 0.876–6.909 | 0.0876 | |||
Both pre-S2 and pre-S1 + pre-S2 del (yes vs. no) | 1.621 | 0.393–6.690 | 0.5042 | |||
All three types of pre-S del (yes vs. no) | 1.352 | 0.694–2.632 | 0.3755 | |||
Del spanning the pre-S1 gene segment (yes vs. no) | 1.165 | 0.673–2.015 | 0.5859 | |||
Del spanning the pre-S2 gene segment (yes vs. no) | 1.825 | 1.058–3.149 | 0.0307 * | 2.114 | 1.203–3.714 | 0.0092 ** |
Characteristics | Univariate Analysis | Multivariate Analysis | ||||
---|---|---|---|---|---|---|
HR | 95% CI | p Value | HR | 95% CI | p Value | |
Child–Pugh cirrhosis score (B/C vs. A) | 2.189 | 1.195–4.013 | 0.0112 * | 2.148 | 1.113–4.147 | 0.0227 *,c |
2.123 | 1.114–4.045 | 0.0221 *,d | ||||
AJCC TNM stage (IIIA/IIIB/IIIC/IVA/IVB vs. I/II) | 4.048 | 2.123–7.719 | <0.0001 *** | 3.225 | 1.600–6.500 | 0.0011 **,c |
3.207 | 1.605–6.410 | 0.0010 **,d | ||||
Pre-S1 del percentage a | ||||||
II (>1.895) vs. I (≤1.895) b | 0.752 | 0.342–1.653 | 0.4778 | |||
III (>5.629) vs. I (≤1.895) | 1.116 | 0.524–2.378 | 0.7760 | |||
IV (>28.270) vs. I (≤1.895) | 0.984 | 0.455–2.128 | 0.9670 | |||
Pre-S2 del percentage | ||||||
II (>0.643) vs. I (≤0.643) | 1.091 | 0.479–2.483 | 0.8357 | |||
III (>1.824) vs. I (≤0.643) | 1.272 | 0.560–2.887 | 0.5658 | |||
IV (>13.666) vs. I (≤0.643) | 2.020 | 0.971–4.202 | 0.0600 | |||
Pre-S1 + pre-S2 del percentage | ||||||
II (>0.015) vs. I (≤0.015) | 0.501 | 0.207–1.212 | 0.1251 | |||
III (>0.442) vs. I (≤0.015) | 1.109 | 0.513–2.396 | 0.7927 | |||
IV (>2.975) vs. I (≤0.015) | 1.497 | 0.733–3.058 | 0.2687 | |||
Pre-S1 plus pre-S2 del percentage | ||||||
II (>2.725) vs. I (≤2.725) | 0.873 | 0.391–1.946 | 0.7392 | |||
III (>18.539) vs. I (≤2.725) | 1.029 | 0.474–2.233 | 0.9431 | |||
IV (>49.938) vs. I (≤2.725) | 1.220 | 0.562–2.646 | 0.6152 | |||
Pre-S1 plus pre-S1 + pre-S2 del percentage | ||||||
II (>2.043) vs. I (≤2.043) | 0.805 | 0.361–1.797 | 0.5965 | |||
III (>6.332) vs. I (≤2.043) | 1.035 | 0.483–2.216 | 0.9300 | |||
IV (>44.201) vs. I (≤2.043) | 1.162 | 0.529–2.553 | 0.7093 | |||
Pre-S2 plus pre-S1 + pre-S2 del percentage | ||||||
II (>0.689) vs. I (≤0.689) | 1.112 | 0.490–2.526 | 0.7996 | 1.196 | 0.524–2.732 | 0.6713 |
III (>2.109) vs. I (≤0.689) | 1.102 | 0.477–2.547 | 0.8208 | 1.170 | 0.498–2.744 | 0.7190 |
IV (>24.995) vs. I (≤0.689) | 2.265 | 1.067–4.811 | 0.0333 * | 2.351 | 1.073–5.153 | 0.0328 *c |
IV (>24.995) vs. I/II/III (≤24.995) c | 2.118 | 1.191–3.765 | 0.0106 * | 2.102 | 1.148–3.850 | 0.0161 *d |
All three types of pre-S del percentage | ||||||
II (>2.828) vs. I (≤2.828) | 0.688 | 0.302–1.564 | 0.3720 | |||
III (>21.666) vs. I (≤2.828) | 0.819 | 0.373–1.802 | 0.6202 | |||
IV (>58.035) vs. I (≤2.828) | 1.759 | 0.827–3.739 | 0.1422 |
Characteristics | Univariate Analysis | Multivariate Analysis | ||||
---|---|---|---|---|---|---|
HR | 95% CI | p Value | HR | 95% CI | p Value | |
Child-Pugh cirrhosis score (B/C vs. A) | 2.189 | 1.195–4.013 | 0.0112 * | 2.213 | 1.154–4.244 | 0.0168 * |
AJCC TNM stage (IIIA/IIIB/IIIC/IVA/IVB vs. I/II) | 4.048 | 2.123–7.719 | <0.0001 *** | 3.440 | 1.689–7.005 | 0.0007 *** |
Combined pre-S del a | ||||||
com-II b vs. com-I | ||||||
com-III vs. com-I | 1.294 | 0.585–2.863 | 0.5242 | 1.764 | 0.780–3.986 | 0.1726 |
com-IV vs. com-I | 2.237 | 1.222–4.097 | 0.0091 ** | 2.336 | 1.238–4.408 | 0.0088 ** |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Teng, C.-F.; Li, T.-C.; Huang, H.-Y.; Lin, J.-H.; Chen, W.-S.; Shyu, W.-C.; Wu, H.-C.; Peng, C.-Y.; Su, I.-J.; Jeng, L.-B. Next-Generation Sequencing-Based Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma Predicts Hepatocellular Carcinoma Recurrence. Viruses 2020, 12, 796. https://doi.org/10.3390/v12080796
Teng C-F, Li T-C, Huang H-Y, Lin J-H, Chen W-S, Shyu W-C, Wu H-C, Peng C-Y, Su I-J, Jeng L-B. Next-Generation Sequencing-Based Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma Predicts Hepatocellular Carcinoma Recurrence. Viruses. 2020; 12(8):796. https://doi.org/10.3390/v12080796
Chicago/Turabian StyleTeng, Chiao-Fang, Tsai-Chung Li, Hsi-Yuan Huang, Jia-Hui Lin, Wen-Shu Chen, Woei-Cherng Shyu, Han-Chieh Wu, Cheng-Yuan Peng, Ih-Jen Su, and Long-Bin Jeng. 2020. "Next-Generation Sequencing-Based Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma Predicts Hepatocellular Carcinoma Recurrence" Viruses 12, no. 8: 796. https://doi.org/10.3390/v12080796
APA StyleTeng, C. -F., Li, T. -C., Huang, H. -Y., Lin, J. -H., Chen, W. -S., Shyu, W. -C., Wu, H. -C., Peng, C. -Y., Su, I. -J., & Jeng, L. -B. (2020). Next-Generation Sequencing-Based Quantitative Detection of Hepatitis B Virus Pre-S Mutants in Plasma Predicts Hepatocellular Carcinoma Recurrence. Viruses, 12(8), 796. https://doi.org/10.3390/v12080796