肿瘤坏死因子-α诱导腺样囊性癌细胞凋亡中p53和bcl-2基因的表达

Expression of P53 and bcl-2 proteins in the cell apoptosis of adenoid cystic carcinoma induced by Tumor Necrosis Factor-alpha(TNF-α)

目的 探讨肿瘤坏死因子 (TumorNecrosisFactor -alpha ,TNF -α)诱导体外人涎腺腺样囊性癌(Salivaryadenoidcysticcarcinoma ,SACC)细胞凋亡过程中P5 3与bcl- 2蛋白的表达 ,以揭示TNF -α诱导的凋亡与癌基因表达间的相互关系。方法 SACC细胞于RPMI 16 40培养液 (含 10 %胎牛血清 )中 ,37 C恒温孵育。实验组用TNF -α处理 ;对照组只加入胎牛血清作为对照。实验组与对照组均分别于 2 4、48、72小时取出细胞爬片 ,免疫组化 (SP法 )检测P5 3与bcl- 2蛋白的表达情况。结果 经TNF -α处理的SACC细胞 2 4小时后 ,细胞出现胞体变小 ,胞核固缩的凋亡早期的形态学改变。P5 3和bcl - 2蛋白表达检测 ,经RelativeIdentifiedDistributionunit(RIDIT)分析表明 ,在 2 4、48、72小时 ,实验组P5 3蛋白的高染率分别为 5 %、2 5 %、5 5 % ,bcl- 2蛋白为 15 %、2 0 %、35 % ;在 48、72小时 ,实验组与对照组均存在显著性差异 (P <0 .0 1) ;且P5 3蛋白的高染率与相应时间段的细胞凋亡率有相关性 (P <0 .0 1) ,而bcl- 2蛋白的高染率与相应时间段的细胞凋亡率无相关关系 (P >0 .0 5 )。结论 在TNF -α诱导SACC细胞凋亡的早期阶段 ,P5 3及bcl- 2蛋白表达均增强。

Objective To study the expression of P53 and bcl-2 proteins in the cell apoptosis of salivary adenoid cystic carcinoma(SACC) induced by Tumor Necrosis Factor-alpha(TNF-α).Methods SACC cells were incubated with RPMI1640 at 37*{C.Cells were treated by TNF-α as experiment group,while cells were added with serum as control group.Both were taken out after 24,48,72 hours and were examined immunohistochemically.Results ①After 24 hours,the cells treated by TNF-α took on morphological changes of apoptosis,while the cells of control group scarcely did.②After 24,48,72hours treated by TNF-α,the staining intensity of either P53 or bcl-2 protein in the experiment group was higher than that of the control group,the high-staining cell rates were 5%,25%,55%(P53 protein),and 15%,20%,35%(bcl1-2 protein) respectively.The data showed that there was significant difference between the experiment group and the control group at both 48 hour and 72 hour (P< 0.05 ).The high-staining rate of either P53 protein was obviously related to the apoptosis rate of SACC cells in the three stages (P< 0.01 ),while there was no relationship between the high-staining rate of bcl-2 protein and the apoptosis rate.Conclusion SACC cells apoptosis can be induced by TNF-α and the rate of apoptosis may be increased.