BACKGROUND Hepatitis B virus(HBV)is a major cause of hepatocellular carcinoma(HCC).HBV DNA can get integrated into the hepatocyte genome to promote carcinogenesis.However,the precise mechanism by which the integrated ...BACKGROUND Hepatitis B virus(HBV)is a major cause of hepatocellular carcinoma(HCC).HBV DNA can get integrated into the hepatocyte genome to promote carcinogenesis.However,the precise mechanism by which the integrated HBV genome promotes HCC has not been elucidated.AIM To analyze the features of HBV integration in HCC using a new reference database and integration detection method.METHODS Published data,consisting of 426 Liver tumor samples and 426 paired adjacent non-tumor samples,were re-analyzed to identify the integration sites.Genome Reference Consortium Human Build 38(GRCh38)and Telomere-to-Telomere Consortium CHM13(T2T-CHM13(v2.0))were used as the human reference genomes.In contrast,human genome 19(hg19)was used in the original study.In addition,GRIDSS VIRUSBreakend was used to detect HBV integration sites,whereas high-throughput viral integration detection(HIVID)was applied in the original study(HIVID-hg19).RESULTS A total of 5361 integration sites were detected using T2T-CHM13.In the tumor samples,integration hotspots in the cancer driver genes,such as TERT and KMT2B,were consistent with those in the original study.GRIDSS VIRUSBreakend detected integrations in more samples than by HIVID-hg19.Enrichment of integration was observed at chromosome 11q13.3,including the CCND1 pro-moter,in tumor samples.Recurrent integration sites were observed in mitochondrial genes.CONCLUSION GRIDSS VIRUSBreakend using T2T-CHM13 is accurate and sensitive in detecting HBV integration.Re-analysis provides new insights into the regions of HBV integration and their potential roles in HCC development.展开更多
Aim: To study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration. Methods: Sperm chromosomes of 14 subjects (5 healthy controls and 9 HB p...Aim: To study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration. Methods: Sperm chromosomes of 14 subjects (5 healthy controls and 9 HB patients, including 1 acute hepatitis B, 2 chronic active hepatitis B, 4 chronic persistent hepatitis B, 2 HBsAg chronic carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free hamster oocytes and human spermatozoa. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes. Results: Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis B. In 9(9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots and the others 2 to 4 signals. The fluorescence intensity showed significant difference among the signal spots. The distribution of signal sites among chromosomes seems to be random. Conclusion: HBV could integrate into human sperm chromosomes. Results suggest that the possibility of vertical transmission of HBV via the germ line to the next generation is present.展开更多
Integration of oncogenic DNA viruses into the human genome is a key step in most virusinduced carcinogenesis.Here,we constructed a virus integration site(VIS)Atlas database,an extensive collection of integration break...Integration of oncogenic DNA viruses into the human genome is a key step in most virusinduced carcinogenesis.Here,we constructed a virus integration site(VIS)Atlas database,an extensive collection of integration breakpoints for three most prevalent oncoviruses,human papillomavirus,hepatitis B virus,and Epstein-Barr virus based on the next-generation sequencing(NGS)data,literature,and experimental data.There are 63,179 breakpoints and 47,411 junctional sequences with full annotations deposited in the VIS Atlas database,comprising 47 virus genotypes and 17 disease types.The VIS Atlas database provides(1)a genome browser for NGS breakpoint quality check,visualization of VISs,and the local genomic context;(2)a novel platform to discover integration patterns;and(3)a statistics interface for a comprehensive investigation of genotypespecific integration features.Data collected in the VIS Atlas aid to provide insights into virus pathogenic mechanisms and the development of novel antitumor drugs.展开更多
INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For...INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).展开更多
Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors a...Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors affect its performance.Methods:A retrospective cohort analysis involving 550 patients was conducted to evaluate whether the HPV integration plays a role in identifying high-grade cervical lesions and cervical cancer.Logistic regression models and area under the curve(AUC)calculations were employed.Results:Our findings revealed that 53.5%of CGB/surgery pairs demonstrated congruent diagnoses,whereas 17.1%showed underestimation and 29.5%overestimation.Furthermore,multivariate logistic regression analysis identified several key predictors for cervical intraepithelial neoplasia(CIN)2+and CIN3+according to surgical pathology.Notably,a CGB confirming CIN2+[odds ratio(OR)=6.0,95%confidence interval(CI):3.9–9.1,P<0.001],high-grade cytology(OR=2.6,95%CI:1.4–4.9,P=0.003),and HPV integration positivity(OR=2.2,95%CI:1.3–3.5,P<0.001)emerged as significant factors for CIN2+.Similarly,for CIN3+identification,CGB confirming CIN2+(OR=5.3,95%CI:3.4–8.3,P<0.001),high-grade cytology(OR=2.6,95%CI:1.5–4.7,P=0.001),and HPV integration positivity(OR=2.0,95%CI:1.3–3.1,P=0.003)were independent predictors.Conclusion:Our study highlights the innovative role of HPV integration testing as a pivotal adjunct to CGB and cytology,offering a comprehensive approach that may enhance the diagnostic precision for high-grade cervical lesions,ultimately achieving more precise management strategies.展开更多
AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, a...AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.展开更多
Hepatocellular carcinoma(HCC)is the fifth most widespread cancer responsible for one fourth of cancer-related deaths globally.Persistent infection with hepatitis B virus(HBV)remains the main cause of HCC summing up to...Hepatocellular carcinoma(HCC)is the fifth most widespread cancer responsible for one fourth of cancer-related deaths globally.Persistent infection with hepatitis B virus(HBV)remains the main cause of HCC summing up to 50%of its causative etiology.Our recent studies,supported by findings from others,uncovered that HBV and its close relative woodchuck hepatitis virus(WHV)integrate into hepatocyte genome almost immediately,hence in minutes after infection.Retrotransposons and genes with translocation potential were found to be frequent sites of HBV insertions,suggesting a mechanism of HBV DNA spread across liver genome from the earliest stages after virus invasion.Many other genes were identified as the sites of early hepadnavirus merges in human hepatocyte-like lines infected de novo with HBV and in natural woodchuck WHV infection model.It was uncovered that head-to-tail joins(HTJs)prevail among the earliest virus-host fusions,implying their formation via the non-homologous-end-joining(NHEJ)pathway.Overlapping homologous junctions resulting from the micro-homology-mediated-overlapping-joining(MHMOJ)were rarely detected.Formation of the initial HTJs coincided with strong induction of reactive oxygen species(ROS)and transient appearance of inducible nitric oxide(iNOS).This was accompanied by cell DNA damage and activation of the poly(ADP-ribose)polymerase 1(PARP1)-mediated host DNA repair machinery,which may explain predominant HTJ format of the first virushost fusions.Identification of initial integration sites and resulting alterations in hepatocyte phenotype may pave a way to discovery of reliable markers of HBV-triggered HCC,including HCC resulting from occult HBV infection.Our research strongly argues that HBV is an ultimate human carcinogen capable of initiation of a pro-oncogenic process immediately after first contact with a susceptible host.展开更多
文摘BACKGROUND Hepatitis B virus(HBV)is a major cause of hepatocellular carcinoma(HCC).HBV DNA can get integrated into the hepatocyte genome to promote carcinogenesis.However,the precise mechanism by which the integrated HBV genome promotes HCC has not been elucidated.AIM To analyze the features of HBV integration in HCC using a new reference database and integration detection method.METHODS Published data,consisting of 426 Liver tumor samples and 426 paired adjacent non-tumor samples,were re-analyzed to identify the integration sites.Genome Reference Consortium Human Build 38(GRCh38)and Telomere-to-Telomere Consortium CHM13(T2T-CHM13(v2.0))were used as the human reference genomes.In contrast,human genome 19(hg19)was used in the original study.In addition,GRIDSS VIRUSBreakend was used to detect HBV integration sites,whereas high-throughput viral integration detection(HIVID)was applied in the original study(HIVID-hg19).RESULTS A total of 5361 integration sites were detected using T2T-CHM13.In the tumor samples,integration hotspots in the cancer driver genes,such as TERT and KMT2B,were consistent with those in the original study.GRIDSS VIRUSBreakend detected integrations in more samples than by HIVID-hg19.Enrichment of integration was observed at chromosome 11q13.3,including the CCND1 pro-moter,in tumor samples.Recurrent integration sites were observed in mitochondrial genes.CONCLUSION GRIDSS VIRUSBreakend using T2T-CHM13 is accurate and sensitive in detecting HBV integration.Re-analysis provides new insights into the regions of HBV integration and their potential roles in HCC development.
文摘Aim: To study the integration of hepatitis B virus (HBV) DNA into sperm chromosomes in hepatitis B patients and the features of its integration. Methods: Sperm chromosomes of 14 subjects (5 healthy controls and 9 HB patients, including 1 acute hepatitis B, 2 chronic active hepatitis B, 4 chronic persistent hepatitis B, 2 HBsAg chronic carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free hamster oocytes and human spermatozoa. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes. Results: Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis B. In 9(9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots and the others 2 to 4 signals. The fluorescence intensity showed significant difference among the signal spots. The distribution of signal sites among chromosomes seems to be random. Conclusion: HBV could integrate into human sperm chromosomes. Results suggest that the possibility of vertical transmission of HBV via the germ line to the next generation is present.
基金supported by the National Science and Technology Major Project of the Ministry of Science and Technology of China(Grant No.2018ZX10301402)the National Natural Science Foundation of China(Grant No.81761148025)+6 种基金the Major Projects of Guangzhou Science and Technology Bureau,China(Grant No.201704020093)the National Ten Thousands Plan for Young Top Talents,Young Pearl River Scholar,International Cooperation and Exchange Projects of the National Natural Science Foundation of China(China&Russian)(Grant No.17-54-80078)the Major Projects of Wuhan Municipal Health Commission of China(Grant No.WX19M02)the Major Projects of Hubei Provincial Health Commission of China(Grant No.WJ2019H312)the Dongguan Social Development Key Project of China(Grant No.202050715007221)the Regional Joint Fund Project of Guangdong Basic and Applied Basic Research Foundation of China(Regional Cultivation Project,Grant No.2021B1515140063)the Dongguan Social Development Key Project of China(Grant No.20221800905772).
文摘Integration of oncogenic DNA viruses into the human genome is a key step in most virusinduced carcinogenesis.Here,we constructed a virus integration site(VIS)Atlas database,an extensive collection of integration breakpoints for three most prevalent oncoviruses,human papillomavirus,hepatitis B virus,and Epstein-Barr virus based on the next-generation sequencing(NGS)data,literature,and experimental data.There are 63,179 breakpoints and 47,411 junctional sequences with full annotations deposited in the VIS Atlas database,comprising 47 virus genotypes and 17 disease types.The VIS Atlas database provides(1)a genome browser for NGS breakpoint quality check,visualization of VISs,and the local genomic context;(2)a novel platform to discover integration patterns;and(3)a statistics interface for a comprehensive investigation of genotypespecific integration features.Data collected in the VIS Atlas aid to provide insights into virus pathogenic mechanisms and the development of novel antitumor drugs.
基金This work was supported by Projects of Tackling Key Problems in ScienceTechnology from the State Science+2 种基金Technology Ministry (TJ99-LA01) Shanghai ScienceTechnology Commission (994919033 )
文摘INTRODUCTIONHepatitis B virus (HBV) belongs to the group ofhepatovirus, a major pathogen of human acute andchronic hepatitis B[1 4], which has a very closeassociation with human hepatocellular carcinoma(HCC)[5-8], For example, a statistical data from ahospital in Shanghai showed that 80% of HCCpatients were positive for HBsAg ( personalcommunication).
文摘Objective:Our research aims to evaluate the diagnostic accuracy of colposcopy-guided biopsy(CGB)in detecting high-grade cervical lesions and explore how human papilloma virus(HPV)integration status and other factors affect its performance.Methods:A retrospective cohort analysis involving 550 patients was conducted to evaluate whether the HPV integration plays a role in identifying high-grade cervical lesions and cervical cancer.Logistic regression models and area under the curve(AUC)calculations were employed.Results:Our findings revealed that 53.5%of CGB/surgery pairs demonstrated congruent diagnoses,whereas 17.1%showed underestimation and 29.5%overestimation.Furthermore,multivariate logistic regression analysis identified several key predictors for cervical intraepithelial neoplasia(CIN)2+and CIN3+according to surgical pathology.Notably,a CGB confirming CIN2+[odds ratio(OR)=6.0,95%confidence interval(CI):3.9–9.1,P<0.001],high-grade cytology(OR=2.6,95%CI:1.4–4.9,P=0.003),and HPV integration positivity(OR=2.2,95%CI:1.3–3.5,P<0.001)emerged as significant factors for CIN2+.Similarly,for CIN3+identification,CGB confirming CIN2+(OR=5.3,95%CI:3.4–8.3,P<0.001),high-grade cytology(OR=2.6,95%CI:1.5–4.7,P=0.001),and HPV integration positivity(OR=2.0,95%CI:1.3–3.1,P=0.003)were independent predictors.Conclusion:Our study highlights the innovative role of HPV integration testing as a pivotal adjunct to CGB and cytology,offering a comprehensive approach that may enhance the diagnostic precision for high-grade cervical lesions,ultimately achieving more precise management strategies.
基金Supportod ty the State Key Basic Research Program Grant G1998051211 the Fund for Leading Specialty of Shanghai Metropolitan Bureau of Public Health.
文摘AIM: To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms. METHODS: Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied. RESULTS: Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10). CONCLUSION: Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.
基金supported by an operating grant(PIN 22346)from the Cancer Research Society Inc.,Canada and the Environmental Cancer Fund-Read for the Cure,Canada and in part by an operating grant(PJT-153001)from the Canadian Institutes of Health Research awarded to Michalak TI.
文摘Hepatocellular carcinoma(HCC)is the fifth most widespread cancer responsible for one fourth of cancer-related deaths globally.Persistent infection with hepatitis B virus(HBV)remains the main cause of HCC summing up to 50%of its causative etiology.Our recent studies,supported by findings from others,uncovered that HBV and its close relative woodchuck hepatitis virus(WHV)integrate into hepatocyte genome almost immediately,hence in minutes after infection.Retrotransposons and genes with translocation potential were found to be frequent sites of HBV insertions,suggesting a mechanism of HBV DNA spread across liver genome from the earliest stages after virus invasion.Many other genes were identified as the sites of early hepadnavirus merges in human hepatocyte-like lines infected de novo with HBV and in natural woodchuck WHV infection model.It was uncovered that head-to-tail joins(HTJs)prevail among the earliest virus-host fusions,implying their formation via the non-homologous-end-joining(NHEJ)pathway.Overlapping homologous junctions resulting from the micro-homology-mediated-overlapping-joining(MHMOJ)were rarely detected.Formation of the initial HTJs coincided with strong induction of reactive oxygen species(ROS)and transient appearance of inducible nitric oxide(iNOS).This was accompanied by cell DNA damage and activation of the poly(ADP-ribose)polymerase 1(PARP1)-mediated host DNA repair machinery,which may explain predominant HTJ format of the first virushost fusions.Identification of initial integration sites and resulting alterations in hepatocyte phenotype may pave a way to discovery of reliable markers of HBV-triggered HCC,including HCC resulting from occult HBV infection.Our research strongly argues that HBV is an ultimate human carcinogen capable of initiation of a pro-oncogenic process immediately after first contact with a susceptible host.
