8-Br-cAMP对Hela细胞wtp53,mtp53,c-myc和iNOS基因表达的影响

Effects of 8-Br-cAMP on gene expression of wtp53, mtp53, c-myc and iNOS in Hela cells

目的 :探讨 8 Br cAMP对Hela细胞癌基因和抑癌基因等表达的影响。方法 :将培养的Hela细胞分为 3组 :① 8 Br cAMP组 (Br组 ) :加至 8 Br cAMP终浓度为 2× 10 -5mol/L ;②阳性对照组 (60 Co组 ) :给予60 Co照射量 4Gy ,72 0s;③阴性对照组 (C组 ) :仅加含体积分数为 10 %胎牛血清的DMEM培养液。各组细胞均培养 4 8h后 ,将 1× 10 7ml-1细胞悬液滴加至预先处理过的硝酸纤维素膜和载玻片上。应用原位杂交和完整细胞原位斑点印迹技术分别检测 3组Hela细胞中野生型p5 3(wtp5 3)、突变型p5 3(mtp5 3)、c myc和iNOS基因的表达。结果 :8 Br cAMP组与阳性对照组相比 ,差异无统计学意义 ,P >0 .0 5 ;与阴性对照组相比 ,上调wtp5 3和iNOS基因的表达 ,同时下调mtp5 3和cmyc基因的表达 ,P <0 .0 1。结论 :结果提示 8 Br

Aim: To study the effects of 8 Br cAMP on oncogene and anti oncogene in human Hela cells. Methods:2×10 -5 mol/L 8 Br cAMP as a final concentration was added into the cultured Hela cells. The cultured Hela cells treated by 60 Co radiation with 4 Gy for 720 sec as a positive control, whereas the cultured Hela cells without 8 Br cAMP was used as the negative control. The cultured Hela cell suspension for each group was blotted onto the pretreated slide and nitrocellulose membrane respectively. The gene expression of wild type p53 (wtp53), mutant type p53(mtp53), c myc, and iNOS were detected by in situ hybridization and RNA dot blot respectively. Results: In Br group the gene expression of wtp53 and iNOS was up regulated, while that of mtp53 and c myc was down regulated compared with group C.Conclusion:The alteration of gene expression of oncogenes and anti oncogenes could be induced by 8 Br cAMP which may be involved in differentiation of Hale cells.