Effect On Apoptotic Related Gene Expression Of HeLa Cell With CD59-transfected Gene By He-Ne Laser Irradiation

Objective To construct a eukaryotic expression system for CD59. To get HeLa cell lines with the highly expression of CD59. To discuss the effect on the Fas, Bcl-2 and NF-κB expression in HeLa Cell and HeLa Cell with CD59-transfected Gene in vitro and in vivo induced by He-Ne laser and study the resistance of CD59 gene to apoptotic signals of tumors induced by He-Ne Laser.Methods The recombinant PALTER-MAX plasmid containing CD59 cDNA and PCDNA plasmid were co-transfected into HeLa cell by cation lipoid mediating method. The positive clones were selected by G418, and highly expressing clones were more selected by ELISA, and get stable highly expressing HeLa cell lines. Stable highly expressing HeLa cell lines were more detected by in situ hybridization. HeLa cell and HeLa cell with CD59-transfected gene in vitro was irradiated by He-Ne laser. The expression of Fas, bcl-2 and NF-κB in the cells was determined with immunohiStochemistry technique. HeLa cell and HeLa cell with CD59-transfected gene were injected into the back of mice respectively. After 24h of the injection, mice were further treated with laser therapy. The mice were killed in thesixteen day and get the thymuses, spleens. The effect of He-Ne laser on thymus,spleen was also evaluated by MTT method. The expression of Fas, Bcl-2 and NF-κB in tumor was determined with immunohistochemistry technique.Result A eukaryotic expression system for highly expression CD59 was successfully constructed. The recombinant PALTER-MAX plasmid containing CD59 cDNA and PCDNA plasmid were co-transfected into HeLa cell by cation lipoid mediating method. Highly expressing clones were selected. by ELISA and in situ hybridization. HeLa cell and HeLa cell with CD59-transfected gene in vitro was irradiated by He-Ne laser. Compared with the control group (A group), the Fas expression of HeLa cell by He-Ne laser irradiation (B group) is significantly increased. Bcl-2, NF-κB and CD59 expression of B group is significantly reduced compared with A group. Compared with B group, the Fas expression of HeLa cell with CD59-transfected gene(C group) is reduced and Bcl-2, NF-κB, CD59 expression of C group is significantly increased. HeLa cell. and HeLa cell with CD59-transfected gene were injected into the back of mice. The mice were treated with He-Ne laser. The mice were killed in the sixteen day and get the thymus,, spleens. The thymus; spleens were grinded and the killings of NK cell were ditected by MTT method. He-Ne laser strengthens NK cell to kill K562 cell. A statistically significant increase in the expression level of Fas in the tumors is noted betweenⅠgroup andⅡgroup. No significant difference is detected between the expression levels of Fas betweenⅡgroup andⅢgroup. Compared withⅠgroup, tumor weight, Bcl-2, NF-κB and CD59 expression ofⅡgroup is significantly reduced. Compared WithⅢgroup, tumor weight, Bcl-2, NF-κB and CD59 expression ofⅡgroup are significantly reduced.Conclusion He-Ne laser can induce the apoptotic signal of HeLa lines and inhibit the growth, of the tumor. CD59 can promote the expression of Bcl-2, NF-κB and growth of the tumor.