转染分离酶cDNA对spc细胞染色体倍性的影响

发布时间：2018-02-27 18:26

本文关键词： spc 细胞分离酶保全素粘合素染色体细胞周期　出处：《第三军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：背景: 染色体是细胞内维持遗传物质稳定和保证其完整、准确传递的基本结构。在有丝分裂时,姐妹染色单体的正确分离既起到保证遗传物质准确分配的作用,又是分裂细胞从后期向末期过渡所必需的;而姐妹染色单体不分离则是产生非整倍体细胞的重要原因。姐妹染色单体分离主要受到分离酶(separase/esp1)及相关蛋白的调控。通常情况下分离酶受到保全素(securin/PTTG)的抑制而没有活性,当细胞从有丝分裂中期向后期转变时,APC(anaphase-promoting complex, 后期促进复合体)被激活并裂解保全素,具有蛋白酶活性的分离酶裂解粘合姐妹染色单体的粘合素(cohesin),姐妹染色单体分离。染色体数目畸变是肿瘤细胞的特征性表现之一,其发生机制至今未见有深入研究。上述姐妹染色单体分离机制是否参与肿瘤细胞染色体数目畸变过程是一个值得研究的重要课题。目的: 观察上调表达分离酶对spc 细胞染色体倍性的影响。方法: 1.利用脂质体转染技术将分离酶cDNA 转入spc 细胞。 2.利用实时荧光定量PCR 及原位杂交的方法检测分离酶基因表达情况。 3.利用人工计数和FACS 检测spc 细胞染色体倍性的变化。结果: 1.转入分离酶cDNA 后,检测到分离酶的上调表达; 2.转染分离酶cDNA 后spc 细胞染色体众数减少。 3.转染分离酶cDNA 后spc 细胞的凋亡受到抑制。结论: 上调表达分离酶基因可以导致spc 细胞的染色体数目显著减少。有趣的是,我们还发现,上调表达分离酶基因后spc 细胞的凋亡受到抑制。
[Abstract]:Background:. Chromosomes are basic structures that maintain the stability of genetic material and ensure its integrity and accurate transmission. In mitosis, the correct separation of sister chromatids plays a role in ensuring the accurate distribution of genetic material. It is also necessary for the transition of mitotic cells from anaphase to late stage, and the non-separation of sister chromatid is an important cause of aneuploidy cells. Sister chromatid separation is mainly regulated by the isolating enzyme separation / esp1) and related proteins. In general, the isolase is inhibited and not active by the preservative securin / PTTG. APCanaphase-promoting (anaphase complex) was activated and lysed preservation, protease-active isolating enzyme cleavage binding sister chromatid adhesinine, sister chromatid separation when cells changed from metaphase to anaphase. Chromosome number aberration is one of the characteristic manifestations of tumor cells. So far, no further studies have been conducted on the mechanism of Sister chromatid segregation. It is an important issue to study whether the above sister chromatid segregation mechanism is involved in the process of chromosome number aberration in tumor cells. Objective:. To observe the effect of up-regulated expression isolase on chromosome ploidy of spc cells. Methods:. 1. The isolase cDNA was transfected into spc cells by liposome transfection. 2. The expression of isolase gene was detected by real-time fluorescence quantitative PCR and in situ hybridization. 3. The changes of chromosome ploidy in spc cells were detected by artificial count and FACS. Results:. 1. The up-regulated expression of isolase cDNA was detected. 2. The chromosome mode of spc cells decreased after transfection of cDNA. 3. The apoptosis of spc cells was inhibited after transfection of cDNA. Conclusion:. Upregulating the expression of isolase gene can significantly reduce the number of chromosomes in spc cells. Interestingly, we also found that the apoptosis of spc cells was inhibited after up-regulation of the expression of isolase gene.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R394

【共引文献】