Investigation Of Inhibition Of Liver Metastasis Of Colon Carcinoma On The Nude Mice Models With Wild Type P53 Gene Combined The System Of TK/GCV And CD/5-Fc

Objective To explore the inhibition of WTp53 gene combined with TK/GCV, CD/5-FC system on liver metastasis model of colon carcinomaMethods pCMV-p53 plasmid was transferred into colon adenocarcinoma cell line SW480 using Lipofectamine ^TM2000 Reagent, The SW480 cells were infected with the concentrated virus containing the double suicide genes, After G418 selection, SW480/p53 and SW480/TK-CD were acquired. RT-PCR confirmed extraneous gene integration and expression. 48 nude mice were divided into 6 groups at random, and each group had 8 nude mice. Nude mice liver metastasis model of colon carcinoma was established by method of spleen injection . The first group was control group, which included the mice injected SW480 cell alone in spleen. The second group referred to the mice injected SW480/p53 cell in spleen. The mice in the other three groups were all injected SW480/TK-CD cells in the spleen.And GVC, 5-Fc, GVC+5F-c were injected into the peritoneal cavity of the mice in the third group,the fourth group and the fifth group respectively. Different from the fifth group, the mice of the sixth group were injected SW480/p53 and SW480/TK-CD in spleen at an equal ratio, not injecting SW480/TK-CD cell alone. Nude mice common status, liver metastasis rate, the number of liver metastasis, pathology, the pathological changes of the tumor tissures under the electronic microscope, tumor cell's apoptotic index and life span and other indexes of nude mice in six groups were observed.Results The plasmid pCMV-p53 was transfected into SW480 successfully, After G418 selection, SW480/p53 was acquired. The cell line SW480/TK-CD that expressed CD and TK was acquired after transfecting virus supernatant into SW480 and selecting with G418. 546bp, 1. 31kb and 1. 19kb positive bands were seen through RT-PCR, which showed that the WTp53 gene and double suicide gene were expressed in SW480. Liver metastasis rate of nude mice in every treatment group was lower than that in control group. Compared with control group, average number of liver metastasis decreased, life span of nude mice was prolonged, and the apoptosis rate of cancer cell in liver metastasis increased. The combined genes in the sixth group worked best, and there was mutual effect between WTp53 and the combination of TK/GCV and CD/5-Fc.Conclusions Synergistic effect was acquired combining WTp53 gene with TK/GCV and CD/5-FC system. It efficiently inhibitted the formation of liver metastasis of colon carcinoma.