Objective: To study the role of SV40 early region gene coding product large tumor antigen(Tag) expression and the interaction between Tag and tumor suppressors p53 and pRb in human brain tumorigenesis. Methods: Tag wa...Objective: To study the role of SV40 early region gene coding product large tumor antigen(Tag) expression and the interaction between Tag and tumor suppressors p53 and pRb in human brain tumorigenesis. Methods: Tag was investigated by immunoprecipitation followed by silver staining and Western blot in 65 cases of human brain tumors and 8 cases of normal brain tissues. Tag-p53 and Tag-pRb complexes were screened by immunoprecipitation and Western blot in 18 and 15 Tag positive tumor tissues respectively. Results: SV40 Tag was expressed generally in human brain tumors, its positive rate was 66. 2% (43 /65). However, Eight normal brain tissues were all negative for Tag, there was significant difference between them(P < 0. 05). Tag-p53 complex was detected in all of 18 Tag positive tumors as well as Tag-pRb complex in all of 15 Tag positive tumors. Conclnsion: SV40 Tag expression is associated with human brain tumorigenesis. The inactivation of p53 and pRh due to the formation of Tag-p53 and Tag-pRb complexes is possibly an important mechanism in the etiopathogenesis of human brain tumors.展开更多
Objective: To investigate the expression of SV40 Tag and formation of Tag-p53 and Tag-Rb complexes in Chinese brain tumors. Methods: SV40 large tumor antigen (Tag) were investigated by immunoprecipitation, silver stai...Objective: To investigate the expression of SV40 Tag and formation of Tag-p53 and Tag-Rb complexes in Chinese brain tumors. Methods: SV40 large tumor antigen (Tag) were investigated by immunoprecipitation, silver staining and Western blot in 65 cases of Chinese brain tumors and 8 cases of normal brain tissues. Tag-p53 and Tag-Rb complexes were screened by the same way in 20 and 15 Tag positive tumor tissues respectively. Results: Tag was found in all of 8 ependymomas and 2 choroid plexus papillomas, 90% (9/10) of pituitary adenomas, 73% (11/15) of astrocytomas, 70% (7/10) of meningiomas, 50% (4/8) of glioblastoma multiform, 33% (2/6) of medulloblastomas, 5 oligodendrogliomas, 1 pineocytoma and 8 normal brain tissues were negative for Tag. Tag-p53 complex was detected in all of 20 Tag positive tumors as well as Tag-Rb complex in all of 15 Tag positive tumors. Conclusion: SV40 Tag is not only expressed in human brain tumors, but also it can form specific complexes with tumor suppressors p53 and Rb. SV40 is correlated to human brain tumorigenesis. The inactivation of p53 and Rb due to the formation of Tag-p53 and Tag-Rb complexes is possibly an important mechanism in the etiopathogenesis of human brain tumors.展开更多
目的建立稳定的SV40感染滴度测定方法,制备高滴度SV40,用于生物制品病毒清除/灭活工艺的验证。方法通过分析不同细胞感染SV40后出现病变的时间、病变程度及产毒量,确定SV40敏感细胞株。分析维持液、细胞培养时间、病毒吸附时间等对病毒...目的建立稳定的SV40感染滴度测定方法,制备高滴度SV40,用于生物制品病毒清除/灭活工艺的验证。方法通过分析不同细胞感染SV40后出现病变的时间、病变程度及产毒量,确定SV40敏感细胞株。分析维持液、细胞培养时间、病毒吸附时间等对病毒滴定测定的影响,建立SV40滴度测定的方法。并分析病毒感染后不同时间及细胞不同部位SV40滴度的差异,制备大量的高滴度SV40。结果与Vero、Vero76及VeroE6细胞相比,CV-1细胞对SV40高度敏感,细胞病变出现时间最早,病变最明显,产毒量最高。SV40滴度与病毒接种前细胞的培养时间、细胞接种量和维持液无明显相关,但吸附时间对病毒滴度有一定的影响。病毒的最佳吸附时间为120 min。接种病毒后48 h收集细胞沉淀,所获得的SV40滴度最高,平均为8.81 lg CCID50/ml。结论已建立了稳定的SV40滴度测定方法,并制备了高滴度SV40,为病毒清除/灭活工艺验证研究奠定了基础。展开更多
基金National Natural Science Foundation of China!No.39470724
文摘Objective: To study the role of SV40 early region gene coding product large tumor antigen(Tag) expression and the interaction between Tag and tumor suppressors p53 and pRb in human brain tumorigenesis. Methods: Tag was investigated by immunoprecipitation followed by silver staining and Western blot in 65 cases of human brain tumors and 8 cases of normal brain tissues. Tag-p53 and Tag-pRb complexes were screened by immunoprecipitation and Western blot in 18 and 15 Tag positive tumor tissues respectively. Results: SV40 Tag was expressed generally in human brain tumors, its positive rate was 66. 2% (43 /65). However, Eight normal brain tissues were all negative for Tag, there was significant difference between them(P < 0. 05). Tag-p53 complex was detected in all of 18 Tag positive tumors as well as Tag-pRb complex in all of 15 Tag positive tumors. Conclnsion: SV40 Tag expression is associated with human brain tumorigenesis. The inactivation of p53 and pRh due to the formation of Tag-p53 and Tag-pRb complexes is possibly an important mechanism in the etiopathogenesis of human brain tumors.
基金This work was supported by China Postdoctoral Science Foundation (No. 1998-23).
文摘Objective: To investigate the expression of SV40 Tag and formation of Tag-p53 and Tag-Rb complexes in Chinese brain tumors. Methods: SV40 large tumor antigen (Tag) were investigated by immunoprecipitation, silver staining and Western blot in 65 cases of Chinese brain tumors and 8 cases of normal brain tissues. Tag-p53 and Tag-Rb complexes were screened by the same way in 20 and 15 Tag positive tumor tissues respectively. Results: Tag was found in all of 8 ependymomas and 2 choroid plexus papillomas, 90% (9/10) of pituitary adenomas, 73% (11/15) of astrocytomas, 70% (7/10) of meningiomas, 50% (4/8) of glioblastoma multiform, 33% (2/6) of medulloblastomas, 5 oligodendrogliomas, 1 pineocytoma and 8 normal brain tissues were negative for Tag. Tag-p53 complex was detected in all of 20 Tag positive tumors as well as Tag-Rb complex in all of 15 Tag positive tumors. Conclusion: SV40 Tag is not only expressed in human brain tumors, but also it can form specific complexes with tumor suppressors p53 and Rb. SV40 is correlated to human brain tumorigenesis. The inactivation of p53 and Rb due to the formation of Tag-p53 and Tag-Rb complexes is possibly an important mechanism in the etiopathogenesis of human brain tumors.
文摘目的建立稳定的SV40感染滴度测定方法,制备高滴度SV40,用于生物制品病毒清除/灭活工艺的验证。方法通过分析不同细胞感染SV40后出现病变的时间、病变程度及产毒量,确定SV40敏感细胞株。分析维持液、细胞培养时间、病毒吸附时间等对病毒滴定测定的影响,建立SV40滴度测定的方法。并分析病毒感染后不同时间及细胞不同部位SV40滴度的差异,制备大量的高滴度SV40。结果与Vero、Vero76及VeroE6细胞相比,CV-1细胞对SV40高度敏感,细胞病变出现时间最早,病变最明显,产毒量最高。SV40滴度与病毒接种前细胞的培养时间、细胞接种量和维持液无明显相关,但吸附时间对病毒滴度有一定的影响。病毒的最佳吸附时间为120 min。接种病毒后48 h收集细胞沉淀,所获得的SV40滴度最高,平均为8.81 lg CCID50/ml。结论已建立了稳定的SV40滴度测定方法,并制备了高滴度SV40,为病毒清除/灭活工艺验证研究奠定了基础。
