维甲酸诱导多种肿瘤细胞凋亡相关基因克隆的研究(英文)

Effects of retinoic acid inducing apoptosis-related gene from multiple carcinoma cells

目的应用基于聚合酶链式反应(PCR)技术的改良消减杂交方法克隆肿瘤凋亡相关基因,验证维甲酸调控生物细胞增殖、分化,诱导多种肿瘤细胞凋亡的作用。方法以全反式维甲酸(all-transretinoicacid)诱导前列腺癌DU-145细胞、早幼粒白血病HL-60细胞和乳腺腺癌MCF-7细胞产生凋亡,在此基础上,采用基于PCR技术的改良消减杂交方法克隆肿瘤细胞凋亡相关基因。结果在维甲酸诱导前列腺癌DU-145细胞、早幼粒白血病HL-60细胞和乳腺腺癌MCF-7细胞产生凋亡过程中,有TNF、泛素、热休克蛋白和C-erbB-2基因参与,并成功克隆出多条可能与凋亡密切相关的未知基因,均被GenBank收录,登录号为AF174394,AF144056,AF141882。结论这种基于PCR技术的改良消减杂交方法对于差异表达基因的克隆是十分有效的。

Aim To clone apoptosis related genes by improved PCR based substractive hybridization,and to testify the effects of retinoic acid on controlling the proliferation and differentiation of cells and inducing apoptosis of multiple carcinoma cells.Methods Apoptosis of DU 145(prostate cancer cell line),HL 60(human promyelocyte cell line)and MCF 7(breast cancer cell line) cells was induced by adding all trans retinoic acid firstly.Then apoptosis related genes were cloned by improved PCR based substractive hybridization.Results TNF, ubiquitin, heat shock proteins(HSP) and C erbB 2 genes participated in the retinoic induced apoptosis procedures in all these cell lines; furthermore, we succeed in cloning some unknown apoptosis related genes, they are all accessed by GeneBank(Genebank Accession Number:AF174394,AF 144056, AF 141882).Conclusion The improved PCR based substractive hybridization is very effective in cloning differently expressed genes.<P>