AIM： To clarify the significance of JC virus （JCV） T-antigen （T-Ag） expression in human gastric cancer. METHODS： We investigated the relationship between TAg detected by immunohistochemistry and Epstein- Barr vi...AIM： To clarify the significance of JC virus （JCV） T-antigen （T-Ag） expression in human gastric cancer. METHODS： We investigated the relationship between TAg detected by immunohistochemistry and Epstein- Barr virus （EBV） infection, microsatellite instability （MSI）, and genetic and epigenetic alterations in gastric cancers. Mutations in the p53,β-catenin, K/MS, BRAF, PIK3CA genes were analyzed by PCR- single strand conformation polymorphism and DNA sequencing. Allelic losses were determined by PCR at 7 microsatellite loci. Aberrant DNA methylation was analyzed by MethyLight assay. RESULTS： JCV T-Ag protein expression was found in 49% of 90 gastric cancer tissues. TAg positivity was not correlated with clinicopathological characteristics. TAg expression was detected in a similar percentage of EBV positive cancers （4 of 9, 44%） and EBV negative cancers （35 of 73, 48%）. TAg expression was detected in a significantly lower percentage of MSI-H cancers （14%） than in non MSI-H cancers （55%, P = 0.005）. TAg expression was detected in a significantly higher percentage of cancers with nuclear/cytoplasmic localization of β-catenin （15 of 21, 71%） than in cancers without （42%, P = 0.018）. p53 mutations were detected in a significantly lower percentage of T-Ag positive cancers （32%） than in TAg negative cancers （57%, P = 0.018）. TAg positive gastric cancers showed a significant increase in the allelic losses and aberrant methylation compared with T-Ag negative gastric cancers （P = 0.008 and P = 0.003）. CONCLUSION： The results suggest that JCV T-Ag is involved in gastric carcinogenesis through multiple mechanisms of genetic and epigenetic alterations.展开更多
AIM:To investigate the expression of the hepatitis B virus(HBV)1.3-fold genome plasmid(pHBV1.3)in an immortalized mouse hepatic cell line induced by SV40T-antigen(SV40T)expression.METHODS:Mouse hepatic cells were isol...AIM:To investigate the expression of the hepatitis B virus(HBV)1.3-fold genome plasmid(pHBV1.3)in an immortalized mouse hepatic cell line induced by SV40T-antigen(SV40T)expression.METHODS:Mouse hepatic cells were isolated from mouse liver tissue fragments from 3-5 d old Kunming mice by the direct collagenase digestion method and cultured in vitro.The pRSV-T plasmid was transfected into mouse hepatic cells to establish an SV40LT-immortalized mouse hepatic cell line.The SV40LT-immortalized mouse hepatic cells were identified and transfected with the pHBV1.3 plasmid.The levels of hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in the supernatant were determined by an electrochemiluminescence immunoassay at 24,48,72 and 96 h after transfection.The expressions of HBsAg and hepatitis B c antigen(HBcAg)in the cells were investigated by indirect immunofluorescence analysis.The presence of HBV DNA replication intermediates in the transfected cells and viral particles in the supernatant of the transfected cell cultures was monitored using the Southern hybridization assay and transmission electronic microscopy,respectively.RESULTS:The pRSV-T plasmid was used to immortalize mouse hepatocytes and an SV40LT-immortalized mouse hepatic cell line was successfully established.SV40LT-immortalized mouse hepatic cells have the same morphology and growth characteristics as primary mouse hepatic cells can be subcultured and produce albumin and cytokeratin-18 in vitro.Immortalized mouse hepatic cells did not show the characteristics of tumor cells,as alpha-fetoprotein levels were comparable(0.58±0.37 vs 0.61±0.31,P=0.37).SV40LTimmortalized mouse hepatic cells were then transfected with the pHBV1.3 plasmid,and it was found that the HBV genome replicated in SV40LT-immortalized mouse hepatic cells.The levels of HBsAg and HBeAg continuously increased in the supernatant after the transfection of pHBV1.3,and began to decrease 72 h after transfection.The expressions of HBsAg and HBcAg were observed in the pHBV1.3-transfected cells.HBV DNA replication intermediates were also observed at72 h after transfection,including relaxed circular DNA,double-stranded DNA and single-stranded DNA.Furthermore,a few 42 nm Dane particles,as well as many22 nm subviral particles with a spherical or filamentous shape,were detected in the supernatant.CONCLUSION:SV40T expression can immortalize mouse hepatic cells,and the pHBV1.3-transfected SV40T-immortalized mouse hepatic cell line can be a new in vitro cell model.展开更多
多发性骨髓瘤(multiple myeloma,MM)是一种不可治愈的血液系统恶性肿瘤,尽管新型蛋白酶体抑制剂、免疫调节剂及CD38单抗等药物的应用显著延长了患者的生存时间,但复发耐药仍难以避免。细胞免疫治疗,特别是嵌合抗原受体(chimeric antigen...多发性骨髓瘤(multiple myeloma,MM)是一种不可治愈的血液系统恶性肿瘤,尽管新型蛋白酶体抑制剂、免疫调节剂及CD38单抗等药物的应用显著延长了患者的生存时间,但复发耐药仍难以避免。细胞免疫治疗,特别是嵌合抗原受体(chimeric antigen receptor,CAR)T细胞疗法的快速发展,极大程度的改变了复发/难治性(relapsed/refractory,R/R)MM患者的治疗现状。FDA目前已批准了2款靶向B细胞成熟抗原(B cell maturation antigen,BCMA)的CAR-T细胞产品,使其用于既往接受过4线及以上治疗的R/R MM患者。随着临床研究的不断深入,靶向GPRC5D(G protein-coupled receptor C class Group 5 member D,G蛋白偶联受体C类第5组成员D)的CAR-T细胞治疗也显示出其独特的优势。除了应用于难治复发的患者,多项临床试验支持CAR-T在MM中治疗线数的前移。本文就CAR-T细胞治疗在MM中开展的关键性临床研究展开综述,旨在为临床应用提供参考。展开更多
基金Supported by Grants-in-Aid for Scientific Research from the Ministry of Education,Culture,Sports,Science and Technology of Japan(Yamamoto H,Imai K and Shinomura Y)Grants-in-Aid for Cancer Research from the Ministry of Health,Laborand Welfare of Japan(Yamamoto H)
文摘AIM： To clarify the significance of JC virus （JCV） T-antigen （T-Ag） expression in human gastric cancer. METHODS： We investigated the relationship between TAg detected by immunohistochemistry and Epstein- Barr virus （EBV） infection, microsatellite instability （MSI）, and genetic and epigenetic alterations in gastric cancers. Mutations in the p53,β-catenin, K/MS, BRAF, PIK3CA genes were analyzed by PCR- single strand conformation polymorphism and DNA sequencing. Allelic losses were determined by PCR at 7 microsatellite loci. Aberrant DNA methylation was analyzed by MethyLight assay. RESULTS： JCV T-Ag protein expression was found in 49% of 90 gastric cancer tissues. TAg positivity was not correlated with clinicopathological characteristics. TAg expression was detected in a similar percentage of EBV positive cancers （4 of 9, 44%） and EBV negative cancers （35 of 73, 48%）. TAg expression was detected in a significantly lower percentage of MSI-H cancers （14%） than in non MSI-H cancers （55%, P = 0.005）. TAg expression was detected in a significantly higher percentage of cancers with nuclear/cytoplasmic localization of β-catenin （15 of 21, 71%） than in cancers without （42%, P = 0.018）. p53 mutations were detected in a significantly lower percentage of T-Ag positive cancers （32%） than in TAg negative cancers （57%, P = 0.018）. TAg positive gastric cancers showed a significant increase in the allelic losses and aberrant methylation compared with T-Ag negative gastric cancers （P = 0.008 and P = 0.003）. CONCLUSION： The results suggest that JCV T-Ag is involved in gastric carcinogenesis through multiple mechanisms of genetic and epigenetic alterations.
基金Supported by Jinan Science and Technology Bureau,Shandong Province,China,No.200705095-4
文摘AIM:To investigate the expression of the hepatitis B virus(HBV)1.3-fold genome plasmid(pHBV1.3)in an immortalized mouse hepatic cell line induced by SV40T-antigen(SV40T)expression.METHODS:Mouse hepatic cells were isolated from mouse liver tissue fragments from 3-5 d old Kunming mice by the direct collagenase digestion method and cultured in vitro.The pRSV-T plasmid was transfected into mouse hepatic cells to establish an SV40LT-immortalized mouse hepatic cell line.The SV40LT-immortalized mouse hepatic cells were identified and transfected with the pHBV1.3 plasmid.The levels of hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in the supernatant were determined by an electrochemiluminescence immunoassay at 24,48,72 and 96 h after transfection.The expressions of HBsAg and hepatitis B c antigen(HBcAg)in the cells were investigated by indirect immunofluorescence analysis.The presence of HBV DNA replication intermediates in the transfected cells and viral particles in the supernatant of the transfected cell cultures was monitored using the Southern hybridization assay and transmission electronic microscopy,respectively.RESULTS:The pRSV-T plasmid was used to immortalize mouse hepatocytes and an SV40LT-immortalized mouse hepatic cell line was successfully established.SV40LT-immortalized mouse hepatic cells have the same morphology and growth characteristics as primary mouse hepatic cells can be subcultured and produce albumin and cytokeratin-18 in vitro.Immortalized mouse hepatic cells did not show the characteristics of tumor cells,as alpha-fetoprotein levels were comparable(0.58±0.37 vs 0.61±0.31,P=0.37).SV40LTimmortalized mouse hepatic cells were then transfected with the pHBV1.3 plasmid,and it was found that the HBV genome replicated in SV40LT-immortalized mouse hepatic cells.The levels of HBsAg and HBeAg continuously increased in the supernatant after the transfection of pHBV1.3,and began to decrease 72 h after transfection.The expressions of HBsAg and HBcAg were observed in the pHBV1.3-transfected cells.HBV DNA replication intermediates were also observed at72 h after transfection,including relaxed circular DNA,double-stranded DNA and single-stranded DNA.Furthermore,a few 42 nm Dane particles,as well as many22 nm subviral particles with a spherical or filamentous shape,were detected in the supernatant.CONCLUSION:SV40T expression can immortalize mouse hepatic cells,and the pHBV1.3-transfected SV40T-immortalized mouse hepatic cell line can be a new in vitro cell model.
文摘多发性骨髓瘤(multiple myeloma,MM)是一种不可治愈的血液系统恶性肿瘤,尽管新型蛋白酶体抑制剂、免疫调节剂及CD38单抗等药物的应用显著延长了患者的生存时间,但复发耐药仍难以避免。细胞免疫治疗,特别是嵌合抗原受体(chimeric antigen receptor,CAR)T细胞疗法的快速发展,极大程度的改变了复发/难治性(relapsed/refractory,R/R)MM患者的治疗现状。FDA目前已批准了2款靶向B细胞成熟抗原(B cell maturation antigen,BCMA)的CAR-T细胞产品,使其用于既往接受过4线及以上治疗的R/R MM患者。随着临床研究的不断深入,靶向GPRC5D(G protein-coupled receptor C class Group 5 member D,G蛋白偶联受体C类第5组成员D)的CAR-T细胞治疗也显示出其独特的优势。除了应用于难治复发的患者,多项临床试验支持CAR-T在MM中治疗线数的前移。本文就CAR-T细胞治疗在MM中开展的关键性临床研究展开综述,旨在为临床应用提供参考。
