论文部分内容阅读
用基因重组技术将插入有反义周期蛋白Ｂ１（ｃｙｃｌｉｎＢ１）基因ｐＸＪ４１－ｎｅｏ质粒，转染人结肠腺癌ＨＴ２９，获得了ＨＴＢ１细胞株。ＨＴＢ１细胞显示出ｃｙｃｌｉｎＢ１ｍＲＮＡ明显的表达抑制。为了探讨ＨＴＢ１细胞生长变慢和它的增殖相关基因表达之间的关系，我们用异硫氰酸胍－酚一步法提取ＨＴＢ１和ＨＴ２９细胞的总ＲＮＡ，并用一些有关的基因探针进行杂交实验。其结果表明：ｃ－ｍｙｃ和ＣＤＫ４在ＨＴＢ１中的表达低于在ＨＴ２９中的表达；而另一方面，ｐ５３、Ｒｂ、ＴＧＦβ和ｃｙｃｌｉｎＤ３在ＨＴＢ１细胞中的表达高于对照组。而ｃｄｃ２基因表达二者维持相似水平。根据研究结果我们发现，转染有反义ｃｙｃｌｉｎＢ１基因的ＨＴＢ１细胞在一组正、负调节基因的协同作用下，导致细胞周期运转的变化。
Using gene recombination technology, the pXJ41-neo plasmid containing the antisense cyclin B1 gene was transfected into human colon adenocarcinoma HT29, and the HTB1 cell line was obtained. HTB1 cells showed marked inhibition of cyclinB1 mRNA expression. In order to investigate the relationship between the slow growth of HTB1 cells and the expression of its proliferation-related genes, we extracted the total RNA of HTB1 and HT29 cells by one-step guanidinium isothiocyanate-phenol method and performed hybridization experiments with some related gene probes. The results showed that the expression of c-myc and CDK4 in HTB1 was lower than that in HT29; on the other hand, the expression of p53, Rb, TGFβ and cyclinD3 in HTB1 cells was higher than that in the control group. The cdc2 gene expression maintained similar levels. Based on the results of the study, we found that HTB1 cells transfected with antisense cyclinB1 gene resulted in changes in cell cycle progression under the synergistic effect of a set of positive and negative regulatory genes.