AIM: To evaluate the possibility of the induction of antitumor immune response by transfecting the colorectal cancer cells with chemokine MCP-3 gene. METHODS: Mouse MCP-3 gene was transduced into mouse colorectal canc...AIM: To evaluate the possibility of the induction of antitumor immune response by transfecting the colorectal cancer cells with chemokine MCP-3 gene. METHODS: Mouse MCP-3 gene was transduced into mouse colorectal cancer cells CMT93 by using of Liposome. G418-resistant clones were selected and the MCP-3 mRNA expression was detected by RT-PCR. The chemotactic activity of MCP-3 in the cell culture supernatant was detected by Chemotaxis assay. The tumorigenicity of wild type CMT93 and CMT93 gene transfectants were detected by in vivo experiments. The immune cell infiltrations in tumor tissue and tumor metastasis were detected histopathologically.RESULTS: MCP-3 mRNA expression was detected by RTPCR in gene-transfected cells (CMT93/MCP-3), but not in control groups. And MCP-3 secreted in the cell culture supernatant possessed chemotatic activity. The results from in vivo experiments showed that the tumorigenicity of CMT93/MCP-3 had not decreased, but the tumors derived from CMT93/MCP-3 cells grew more slowly than those from CMT93 cells (1.021±0.253) cm2 VS(1.769±0.371) cm2,P＜0.05) or CMT93/mock cells (1.021±0.253) cm2 VS(1.680±0.643)cm2, P＜0.05). Histophathological results showed few immune cells infiltrating in the tumor tissue derived from the controls. In the tumor tissue derived from CMT93/MCP-3, infiltrating immune cells increased. In addition, no tumor metastasis was found in all mice inoculated with CMT93/MCP-3 tumor cells. But all mice had tumor metastasis in CMT93 controls and 4 in 5 mice had tumor metastasis in CMT93/mock controls.CONCLUSION: The results suggested that the transfection of chemokine MCP-3 gene could promote the induction of anti-colorectal cancer immunity, but the tumor growth could not be inhibited completely by merely MCP-3 gene transfection.展开更多
Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological ch...Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological characterisitics showed that the RGV isolates belong to genus Ranavirus. The DNA sequence analysis of the 5’end of the major capsid protein(MCP) gene of RGV isolates by PCR amplification produced a 531bp fragment.DNA templates were prepared from cells infected with different RGV isolates and the specific primers designed were based on highly conserved region at the 5′ of the gene encoding Frog virus 3(FV3) MCP, which is the typical species of the genus Ranavirus.The PCR products indicated that the nucleotide sequence of MCP gene of the RGV9506,RGV9807 and RGV9808,showed 99.6%, 99.8% and 98.4% homology respectively with the corresponding region of the MCP gene of FV3.展开更多
文摘AIM: To evaluate the possibility of the induction of antitumor immune response by transfecting the colorectal cancer cells with chemokine MCP-3 gene. METHODS: Mouse MCP-3 gene was transduced into mouse colorectal cancer cells CMT93 by using of Liposome. G418-resistant clones were selected and the MCP-3 mRNA expression was detected by RT-PCR. The chemotactic activity of MCP-3 in the cell culture supernatant was detected by Chemotaxis assay. The tumorigenicity of wild type CMT93 and CMT93 gene transfectants were detected by in vivo experiments. The immune cell infiltrations in tumor tissue and tumor metastasis were detected histopathologically.RESULTS: MCP-3 mRNA expression was detected by RTPCR in gene-transfected cells (CMT93/MCP-3), but not in control groups. And MCP-3 secreted in the cell culture supernatant possessed chemotatic activity. The results from in vivo experiments showed that the tumorigenicity of CMT93/MCP-3 had not decreased, but the tumors derived from CMT93/MCP-3 cells grew more slowly than those from CMT93 cells (1.021±0.253) cm2 VS(1.769±0.371) cm2,P＜0.05) or CMT93/mock cells (1.021±0.253) cm2 VS(1.680±0.643)cm2, P＜0.05). Histophathological results showed few immune cells infiltrating in the tumor tissue derived from the controls. In the tumor tissue derived from CMT93/MCP-3, infiltrating immune cells increased. In addition, no tumor metastasis was found in all mice inoculated with CMT93/MCP-3 tumor cells. But all mice had tumor metastasis in CMT93 controls and 4 in 5 mice had tumor metastasis in CMT93/mock controls.CONCLUSION: The results suggested that the transfection of chemokine MCP-3 gene could promote the induction of anti-colorectal cancer immunity, but the tumor growth could not be inhibited completely by merely MCP-3 gene transfection.
文摘Since 1995,three viral isolates named RGV9506,RGV9807 and RGV9808 associated with frog mortality were isolated from farm raised frogs (Rana grylio virus,RGV) in China.Both ultrastructural morphology and serological characterisitics showed that the RGV isolates belong to genus Ranavirus. The DNA sequence analysis of the 5’end of the major capsid protein(MCP) gene of RGV isolates by PCR amplification produced a 531bp fragment.DNA templates were prepared from cells infected with different RGV isolates and the specific primers designed were based on highly conserved region at the 5′ of the gene encoding Frog virus 3(FV3) MCP, which is the typical species of the genus Ranavirus.The PCR products indicated that the nucleotide sequence of MCP gene of the RGV9506,RGV9807 and RGV9808,showed 99.6%, 99.8% and 98.4% homology respectively with the corresponding region of the MCP gene of FV3.
