The development of esophageal cancer accompanied by the presence of human papillomavirus (HPV) DNA into the host genome. By evaluating the expression of this virus for tumor cell origin and also their cell grows and m...The development of esophageal cancer accompanied by the presence of human papillomavirus (HPV) DNA into the host genome. By evaluating the expression of this virus for tumor cell origin and also their cell grows and migrations, we examined esophageal cancer clonality in the context of intra-tumor heterogeneity. In this research, we have checked the expression of HPV18 E6 and E7 in different single cell clones by the manual cell picking method in the HPV positive esophageal cancer (EC109), EC109 cell line used as a negative control, and Hela cell line used as the positive control. Quantitative real-time PCR (QRT-PCR) was run to detect the expression levels of HPV E6 and E7, Cell Counting Kit-8 (CCK-8) assay was used to examine cell proliferation, invasion assays performed using Costar chambers and wounding assay to study cell migrations in vitro. We investigated the intra-tumor heterogeneity of HPV E6 and E7 in esophageal cancer and the evaluation of the growth and migrations at the clonal level, using 10 single cell clones. In particular clones, C7 & C10 displayed a highly variable expression in both HPV E6 and E7 and weak in four clones (C1, C3, C4, and C9) consequently, the cell invasion, proliferation, and migration increase with increasing the level of HPV expression and inverse. In conclusion, the resulting based on single cell cloning showed the relationship between HPV and cell growth and migration in esophageal cancer. Future study in HPV DNA integration needed to explore the mains specific integration site of HPV DNA in esophageal cancer and molecular monitoring of the HPV for future prevention researches and also effective therapeutic strategies.展开更多
Objective. Human papillomavirus type 16 (HPV 16) has several intratyp ic varian ts, and some are associated with enhanced oncogenic potential. For risk determin ation aswell as for future vaccine development, knowledg...Objective. Human papillomavirus type 16 (HPV 16) has several intratyp ic varian ts, and some are associated with enhanced oncogenic potential. For risk determin ation aswell as for future vaccine development, knowledge about variants is impo rtant. Regarding the geographical distribution of HPV variants and the lack of d ata from Indonesia and Suriname, we studied the prevalence of HPV 16 variants in cervical cancer in these high incidence countries. Data were compared with The Netherlands, a low-risk country. Methods. DNA samples from 74 formalin-fixed p araffin-embedded HPV 16-positive cervical carcinomas from Indonesia (Java, N = 22), Suriname (N = 25), and The Netherlands (N = 27) were amplified using prime rs specific for the E6, E7, and part of the L1 regions. Products were sequenced and analyzed. Results. A specific Javanese variant, with mutations 666A in E7 an d 6826T in L1, was found in 73%of the Indonesian samples, 56%having an additio nal mutation in the E6 open reading frame (ORF; 276G), giving the predicted amin o acid change N58S. This Javanese variant was also found in three Surinamese sam ples, which reflects what could be expected from migration of Javanese people to Surinam. Other non-European variants were identified in Indonesian, Surinamese , and Dutch samples in 14%, 28%, and 19%, respectively. Conclusion. The major ity of the HPV 16-positive cervical cancers in Indonesia are caused by a specif ic intratypic variant that was rarely found before in other countries.展开更多
文摘The development of esophageal cancer accompanied by the presence of human papillomavirus (HPV) DNA into the host genome. By evaluating the expression of this virus for tumor cell origin and also their cell grows and migrations, we examined esophageal cancer clonality in the context of intra-tumor heterogeneity. In this research, we have checked the expression of HPV18 E6 and E7 in different single cell clones by the manual cell picking method in the HPV positive esophageal cancer (EC109), EC109 cell line used as a negative control, and Hela cell line used as the positive control. Quantitative real-time PCR (QRT-PCR) was run to detect the expression levels of HPV E6 and E7, Cell Counting Kit-8 (CCK-8) assay was used to examine cell proliferation, invasion assays performed using Costar chambers and wounding assay to study cell migrations in vitro. We investigated the intra-tumor heterogeneity of HPV E6 and E7 in esophageal cancer and the evaluation of the growth and migrations at the clonal level, using 10 single cell clones. In particular clones, C7 & C10 displayed a highly variable expression in both HPV E6 and E7 and weak in four clones (C1, C3, C4, and C9) consequently, the cell invasion, proliferation, and migration increase with increasing the level of HPV expression and inverse. In conclusion, the resulting based on single cell cloning showed the relationship between HPV and cell growth and migration in esophageal cancer. Future study in HPV DNA integration needed to explore the mains specific integration site of HPV DNA in esophageal cancer and molecular monitoring of the HPV for future prevention researches and also effective therapeutic strategies.
文摘Objective. Human papillomavirus type 16 (HPV 16) has several intratyp ic varian ts, and some are associated with enhanced oncogenic potential. For risk determin ation aswell as for future vaccine development, knowledge about variants is impo rtant. Regarding the geographical distribution of HPV variants and the lack of d ata from Indonesia and Suriname, we studied the prevalence of HPV 16 variants in cervical cancer in these high incidence countries. Data were compared with The Netherlands, a low-risk country. Methods. DNA samples from 74 formalin-fixed p araffin-embedded HPV 16-positive cervical carcinomas from Indonesia (Java, N = 22), Suriname (N = 25), and The Netherlands (N = 27) were amplified using prime rs specific for the E6, E7, and part of the L1 regions. Products were sequenced and analyzed. Results. A specific Javanese variant, with mutations 666A in E7 an d 6826T in L1, was found in 73%of the Indonesian samples, 56%having an additio nal mutation in the E6 open reading frame (ORF; 276G), giving the predicted amin o acid change N58S. This Javanese variant was also found in three Surinamese sam ples, which reflects what could be expected from migration of Javanese people to Surinam. Other non-European variants were identified in Indonesian, Surinamese , and Dutch samples in 14%, 28%, and 19%, respectively. Conclusion. The major ity of the HPV 16-positive cervical cancers in Indonesia are caused by a specif ic intratypic variant that was rarely found before in other countries.
