Effect of pEgr-TNFalpha gene radiotherapy on mice melanoma.
In the present study, we constructed a pEgr-tumour necrosis factor-alpha (TNFalpha) plasmid and investigated its expression properties in B16 cells on exposure to ionizing irradiation and, furthermore, the effect of gene radiotherapy on a melanoma model. Firstly, the recombinant pEgr-TNFalpha plasmid was constructed and transfected into B16 cells with liposomes to investigate its expression properties on exposure to X-irradiation. The melanoma-bearing model was then established and the tumour tissue was injected locally with the pEgr-TNFalpha plasmid and exposed to 20 Gy of X-irradiation. The tumour growth curve at different time points was described. At day 3, TNFalpha transcription in the tumour tissue was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that: (1) the eukaryotic expression vector pEgr-TNFalpha was successfully constructed and transfected into B16 cells; (2) TNFalpha expression was significantly increased in the transfected cells after X-irradiation, in contrast with that of the pCMV-TNFalpha group or the pEgr-TNFalpha group that received 0 Gy of irradiation; (3) after pEgr-TNFalpha gene radiotherapy, tumour growth was significantly slower than in those groups receiving irradiation or gene transfer alone. The TNFalpha concentration in the peripheral blood of tumour-bearing mice that received an injection of pEgr-TNFalpha and 20 Gy of irradiation was higher than that of the control group. Only in pEgr-TNFalpha and pEgr-TNFalpha + 20 Gy groups was TNFalpha messenger RNA (mRNA) detected in the tumour tissue. We conclude that pEgr-TNFalpha gene radiotherapy may significantly inhibit tumour growth, and that the anti-melanoma effect is superior to that of either gene therapy or radiotherapy alone. Our work provides the theoretical basis for further study on the gene radiotherapy of this tumour.
Wu
Congmei
C
The Cell Biology Laboratory, Shantou University Medical College, Shantou 515041, China. meiyuing@hotmail.com
Li
Xiuyi
X
Tian
Mei
M
D000818:Animals; D045744:Cell Line, Tumor / Q000528:radiation effects; D003131:Combined Modality Therapy; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D015316:Genetic Therapy; D008546:Melanoma, Experimental / Q000097:blood / Q000235:genetics / Q000532:radiotherapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D009368:Neoplasm Transplantation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000097:blood / Q000235:genetics / Q000532:radiotherapy*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects
D014409:Tumor Necrosis Factor-alpha