Objective To investigate the clinical features of patients developing hepatocellular carcinoma ( HCC) during anti-hepatitis B virus ( HBV) therapy with nucleos ( t) ide analogues ( NAs) . Methods A total of 542 patients who were diagnosed with HCC for the first time in The Third Affiliated Hospital of Sun Yat-Sen University from January 2008 to September 2014 were enrolled, and they all had chronic HBV infection. According to the presence or absence of standard therapy with NAs, they were divided into antiviral group ( 130 patients) and non-antiviral group ( 412 patients) . A retrospective analysis was performed for their clinical data, including age, sex, family history of tumor, duration of HBV infection, the time when a confirmed diagnosis of liver cirrhosis was made, history of drinking, history of diabetes, history of medication, laboratory parameters, liver pathology, and imaging findings, and these data were compared between the two groups.The t-test was used for comparison of normally distributed continuous data between groups, the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups, and the chi-square test was used for comparison of categorical data between groups. Results Compared with the non-antiviral group, the antiviral group had significant increases in the proportion of patients with liver cirrhosis ( 90. 0% vs 78. 4%, χ2= 8. 528, P = 0. 003) and HBe Ag-positive rate ( 29. 4% vs 18. 5%, χ2= 6. 794, P = 0. 009) .There was a significant difference in the constitution of HBV DNA between the two groups ( χ2= 173. 142, P < 0. 001) , as well as significant differences in alanine aminotransferase, gamma-glutamyl transpeptidase, and alpha-fetoprotein ( all P < 0. 001) . Compared with the non-antiviral group, the antiviral group had a higher proportion of patients with early-or intermediate-stage liver cancer, smaller and fewer cancer lesions, and a lower proportion of patients with vascular invasion or distant metastasis ( all P < 0. 001) . Among the patients in the antiviral group, 80% ( 104/130) developed HCC within 5 years of anti-HBV therapy. Conclusion Some patients with chronic HBV infection who receive long-term NAs as the anti-HBV therapy still develop HCC, especially those with liver cirrhosis, and most of these cases occur within 5 years of anti-HBV therapy. Most patients treated with NAs are diagnosed at the early stage of HCC, suggesting that standard follow-up is of vital importance.
旋覆花(安徽省万生中药饮片有限公司,批号:210201),茜草(衢州南孔中药饮片有限公司,批号:2104087);高脂高果糖高胆固醇饲料(美国Research Diets公司,货号:D09100310);HE染色试剂盒、Masson染色试剂盒、油红O染色试剂盒及羟脯氨酸(Hyp)试剂盒(南京建成生物工程研究所,批号或货号分别为20210720、D026-1-3、D027-1、A030-2);甘油三酯(TG)检测试剂盒(浙江东瓯诊断产品有限公司,货号:R0327);奥贝胆酸(大连美仑生物技术有限公司,货号:MB6084);Trizol试剂(上海生工生物工程股份有限公司,批号:G826KA6730);RNA逆转录试剂盒(加拿大Applied Biological Materials公司,货号:G486);SYBRⓇ Green Pro Taq HS预混型实时荧光定量聚合酶链式反应(qPCR)试剂盒(含ROX)(湖南艾科瑞生物工程有限公司,批号:AG11718)。PCR引物均由上海生工生物工程股份有限公司合成,引物序列见表 1。
首先对肝组织RNA进行抽提,称取50 mg肝组织,分别加入1 mL Trizol进行匀浆、裂解,200 μL三氯甲烷使蛋白和RNA迅速脱离,600 μL异丙醇沉淀RNA,1 mL 75%乙醇及无水乙醇洗脱,吸弃残余乙醇,晾干,加入50 μL DEPC水充分溶解,即得到总RNA。随后对总RNA进行cDNA逆转录。最后将cDNA与SYBR green premix试剂混合后置于VIIA 7型PCR仪中进行qPCR扩增,目标基因的相对表达量以GAPDH标准化,采用2-ΔΔCt计算各肝组织中mRNA相对表达量。
[1]HALEGOUA-DE MARZIO D, HANN HW.Then and now:the progress in hepatitis B treatment over the past 20 years[J].World J Gastroenterol, 2014, 20 (2) :401-413.
[2]YE XQ, GAO FY, SUN L, et al.Improvement of improve the two years survival rate of patients with hepatitis B virus-related primary liver cancer after antiviral treatment[J/CD].Chin J Exp Clin Infect Dis:Electronic Edition, 2016, 10 (3) :304-310. (in Chinese) 叶协琼, 高方媛, 孙乐, 等.抗病毒治疗提高乙型肝炎病毒相关原发性肝癌患者两年生存率[J/CD].中华实验和临床感染病杂志电子版, 2016, 10 (3) :304-310.
[3]LIN CL, KAO JH.Risk stratification for hepatitis B virus related hepatocellular carcinoma[J].J Gastroenterol Hepatol, 2013, 28 (1) :10-17.
[5]YANG HI, YUEN MF, CHAN HL, et al.Risk estimation for hepatocellular carcinoma in chronic hepatitis B (REACH-B) :development and validation of a predictive score[J].Lancet Oncol, 2011, 12 (6) :568-574.
[6]European Association ror the Study of the Liver.EASL clinical practice guidelines:management of chronic hepatitis B virus infection[J].J Hepatol, 2012, 57 (1) :167-185.
[7]WONG GL, WONG VW, CHAN HL.Combination therapy of interferon and nucleotide/nucleoside analogues for chronic hepatitis B[J].J Viral Hepat, 2014, 21 (12) :825-834.
[8] WATANABE T, TOKUMOTO Y, JOKO K, et al.Effects of longterm entecavir treatment on the incidence of hepatocellular carcinoma in chronic hepatitis B patients[J].Hepatol Int, 2016, 10 (2) :320-327.
[9]SINGAL AK, SALAMEH H, KUO YF, et al.Meta-analysis:the impact of oral anti-viral agents on the incidence of hepatocellular carcinoma in chronic hepatitis B[J].Aliment Pharmacol Ther, 2013, 38 (2) :98-106.
[10]HOSAKA T, SUZUKI F, KOBAYASHI M, et al.Long-term entecavir treatment reduces hepatocellular carcinoma incidence in patients with hepatitis B virus infection[J].Hepatology, 2012, 58 (1) :98-107.
[11]LIAW YF, SHEEN IS, LEE CM, et al.Tenofovir disoproxil fumarate (TDF) , emtricitabine/TDF, and entecavir in patients with decompensated chronic hepatitis B liver disease[J].Hepatology, 2011, 53 (1) :62-72.
[12] PAPATHEODORIDIS GV, MANOLAKOPOULOS S, TOULOUMI G, et al.Virological suppression does not prevent the development of hepatocellular carcinoma in HBe Ag-negative chronic hepatitis B patients with cirrhosis receiving oral antiviral (s) starting with lamivudine monotherapy:results of the nationwide HEPNET.Greece cohort study[J].Gut, 2011, 60 (8) :1109-1116.
[13] SINGAL AK, SALAMEH H, KUO YF, et al.Meta-analysis:the impact of oral anti-viral agents on the incidence of hepatocellular carcinoma in chronic hepatitis B[J].Aliment Pharmacol Ther, 2013, 38 (2) :98-106.
[14]Chinese Society of Hepatology and Chinese Society of Infectious Diseases, Chinese Medical Association.The guideline of prevention and treatment for chronic hepatitis B:a 2015 update[J].J Clin Hepatol, 2015, 31 (12) :1941-1960. (in Chinese) 中华医学会肝病学分会, 中华医学会感染病学分会.慢性乙型肝炎防治指南 (2015年更新版) [J].临床肝胆病杂志, 2015, 31 (12) :1941-1960.
[15]Ministry of Health of the People's Republic of China.Diagnosis, management, and treatmen of hepatocellular carcinoma (V2011) [J].J Clin Hepatol, 2011, 27 (11) :1141-1159. (in Chinese) 中华人民共和国卫生部.原发性肝癌诊疗规范 (2011年版) [J].临床肝胆病杂志, 2011, 27 (11) :1141-1159.
[16]Biliary Tract Group, Chinese Society of Surgery, Chinese Medical Association Guidelines for the diagnosis and treatment of hepatolithiasis[J].Chin J Intect Dis, 2001, 40 (1) :56-62. (in Chinese) 中华医学会传染病与寄生虫病学分会, 肝病学分会.病毒性肝炎防治方案[J].中华传染病杂志, 2001, 40 (1) :56-62.
[17] SHEN YC, HSU C, CHENG CC, et al.A critical evaluation of the preventive effect of antiviral therapy on the development of hepatocellular carcinoma in patients with chronic hepatitis C or B:a novel approach by using meta-regression[J].Oncology, 2012, 82 (5) :275-289.
Dai ShuYang, Huang Xing, Yang Tian, Shen Feng. Research advances in proton beam therapy for hepatocellular carcinoma[J]. J Clin Hepatol, 2013, 29(10): 744-747. DOI: 10.3969/j.issn.1001-5256.2013.10.006.
Dai ShuYang, Huang Xing, Yang Tian, Shen Feng. Research advances in proton beam therapy for hepatocellular carcinoma[J]. J Clin Hepatol, 2013, 29(10): 744-747. DOI: 10.3969/j.issn.1001-5256.2013.10.006.