Tumor growth inhibition effect of hIL-6 on colon cancer cells transfected with the target gene by retroviral vector.
AIM:To observe the tumor inhibitory effects by transfecting IL-6 cDNA into colon cancer cell line HT-29 with retroviral vector pZIP cDNA.METHODS:Human IL-6 gene was reconstructed in retrovirus vector and transfected into incasing cells PA317 by lipofectamine mediated method, the clones of the cells transferred with hIL-6 were selected by G418,and targeted HT-29 cells were infected with the virus granules secreted from PA317 and also selected by G418.Test gene transcription and expression level by hybridization, ELISA and MTT assay,etc. Analyze tumor inhibitory effects according to the cell growth curve, plating forming rate and tumorigenicity in nude mice.RESULT:Successfully constructed and transfected recombinant expressing vectors pZIPIL-6 cDNA and got positive transfected cell lines. The colon cancer cell line (HT-29 IL-6) transfected with the hIL-6 gene by retroviral vector was estab-lished. The log proliferation period and the doubling time of this cell line was between 4 to 7 days and 2.5 days according to the direct cell count, the cell proliferation was obviously inhibited with MTT assay, the plating inhibitory rate was 50% by plating efficiency test. When HT-29 IL-6 cells were inoculated into the nude mice subcutaneously, carcinogenic activity of the solid tumor was found superior to the control group and the size of tumor was not significantly enlarged. Injection of combination virus fluid containing IL-6 gene into transplantation tumors could inhibit the growth and development of the tumor.CONCLUSION:IL-6 could inhibit the growth and proliferation of colon cancer cells by retroviral vector-mediated transduction.
Xiao
Bing
B
Jing
Bo
B
Zhang
Ya-Li
YL
Zhou
Dian-Yuan
DY
Zhang
Wan-Dai
WD