Suppression of FasL expression in tumor cells and preventing tumor necrosis factor-induced apoptosis by adenovirus 14.7K is an effective escape mechanism for immune cells.
To elucidate if the Fas/FasL signal pathway participates in the immune escape of tumor cells, and if contemporary Fas/FasL and tumor necrosis factor (TNF))-induced apoptosis is better for immune cell survival than just blocking Fas/FasL-induced apoptotic signal. FasL expression in mouse H22 hepatocellular cancer cells was suppressed by the siRNA technique. The wild-type Ad5 14.7K gene was amplified by polymerase chain reaction and transduced into Jurkat T-cells. Apoptosis of target Jurkat cells was detected by flow cytometry. TNF-alpha in the culture supernatant of H22 cells by ELISA was seen. FasL and 14.7K gene expression in stably transfected or transduced clones were determined by Western blotting. As a result, FasL expression in H22 cells was down-regulated after stable transfection with a plasmid encoding antisense FasL cDNA. Down-regulation of FasL expression in H22 cells had no effect on tumor growth in vitro. There was an apparent decrease in the number of apoptotic Jurkat T-cells after coculture with transfected H22 cells, relative to coculture with FasL-expressing untransfected cells. Compared with untransduced Jurkat cells, apoptotic rates in 14.7K-transduced Jurkat cells were significantly reduced in three different E/T ratios (P < 0.01), respectively. We conclude that Fas/FasL signal pathway participates in the immune escape of tumor cells by inducing immune cells apoptosis. Reducing the expression of FasL in tumor cells can decrease the apoptotic rate of immune cells, further blocking the apoptotic signal pathway of immune cells by preventing TNF-induced apoptosis can increase the survival of immune cells.
Zhang
Jiangguo
J
Department of Morphology, Medical College of Huzhou Teachers No. 1, Xueshi Road, Huzhou 313000, China. jg-Zhangyc@hotmail.com
Xu
Gaosi
G
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000276:immunology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D015536:Down-Regulation; D053222:Fas Ligand Protein / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D016372:RNA, Antisense / Q000494:pharmacology; D015398:Signal Transduction; D014161:Transduction, Genetic; D014162:Transfection; D019139:Tumor Escape; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D053222:Fas Ligand Protein; D016372:RNA, Antisense; D014409:Tumor Necrosis Factor-alpha