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  本文關(guān)鍵詞： spc 細(xì)胞 分離酶 保全素 粘合素 染色體 細(xì)胞周期　出處：《第三軍醫(yī)大學(xué)》2005年碩士論文　論文類型：學(xué)位論文

【摘要】：背景: 染色體是細(xì)胞內(nèi)維持遺傳物質(zhì)穩(wěn)定和保證其完整、準(zhǔn)確傳遞的基本結(jié)構(gòu)。在有絲分裂時(shí),姐妹染色單體的正確分離既起到保證遺傳物質(zhì)準(zhǔn)確分配的作用,又是分裂細(xì)胞從后期向末期過(guò)渡所必需的;而姐妹染色單體不分離則是產(chǎn)生非整倍體細(xì)胞的重要原因。 姐妹染色單體分離主要受到分離酶(separase/esp1)及相關(guān)蛋白的調(diào)控。通常情況下分離酶受到保全素(securin/PTTG)的抑制而沒有活性,當(dāng)細(xì)胞從有絲分裂中期向后期轉(zhuǎn)變時(shí),APC(anaphase-promoting complex, 后期促進(jìn)復(fù)合體)被激活并裂解保全素,具有蛋白酶活性的分離酶裂解粘合姐妹染色單體的粘合素(cohesin),姐妹染色單體分離。 染色體數(shù)目畸變是腫瘤細(xì)胞的特征性表現(xiàn)之一,其發(fā)生機(jī)制至今未見有深入研究。上述姐妹染色單體分離機(jī)制是否參與腫瘤細(xì)胞染色體數(shù)目畸變過(guò)程是一個(gè)值得研究的重要課題。 目的: 觀察上調(diào)表達(dá)分離酶對(duì)spc 細(xì)胞染色體倍性的影響。 方法: 1.利用脂質(zhì)體轉(zhuǎn)染技術(shù)將分離酶cDNA 轉(zhuǎn)入spc 細(xì)胞。 2.利用實(shí)時(shí)熒光定量PCR 及原位雜交的方法檢測(cè)分離酶基因表達(dá)情況。 3.利用人工計(jì)數(shù)和FACS 檢測(cè)spc 細(xì)胞染色體倍性的變化。 結(jié)果: 1.轉(zhuǎn)入分離酶cDNA 后,檢測(cè)到分離酶的上調(diào)表達(dá); 2.轉(zhuǎn)染分離酶cDNA 后spc 細(xì)胞染色體眾數(shù)減少。 3.轉(zhuǎn)染分離酶cDNA 后spc 細(xì)胞的凋亡受到抑制。 結(jié)論: 上調(diào)表達(dá)分離酶基因可以導(dǎo)致spc 細(xì)胞的染色體數(shù)目顯著減少。有趣的是,我們還發(fā)現(xiàn),上調(diào)表達(dá)分離酶基因后spc 細(xì)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.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R394

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

1 王臺(tái),丁兆軍;減數(shù)分裂及其基因研究進(jìn)展[J];科學(xué)通報(bào);2002年04期

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本文編號(hào)：1543820