人端粒酶逆轉(zhuǎn)錄酶基因轉(zhuǎn)染U2OS細胞對其基質(zhì)金屬蛋白酶-2活性的影響
發(fā)布時間:2018-10-19 12:56
【摘要】: 目的:觀察hTERT基因轉(zhuǎn)染U2OS細胞對其基質(zhì)金屬蛋白酶2和9的影響,探討hTERT基因和端粒酶的相關(guān)性。 方法:RT-PCR和TRAP-ELISA法檢測和明確對U2OS細胞hTERT基因的轉(zhuǎn)染效果和轉(zhuǎn)染陽性細胞克隆的篩選;明膠酶譜法和RT-PCR檢測轉(zhuǎn)染hTERT基因?qū)毎鸐MP-2,MMP-9表達和活性的影響。 結(jié)果:hTERT基因轉(zhuǎn)染U2OS細胞后,細胞成功的出現(xiàn)hTERT mRNA和明顯的端粒酶活性(p0.05),使該細胞分泌酶原型MMP-(272KD)降低(p0.05),MMP-9完全消失。 結(jié)論:hTERT穩(wěn)定轉(zhuǎn)染U2OS細胞克隆株,使細胞分泌酶原型MMP-9完全消失、酶原型MMP-2明顯減少。但MMP-2在mRNA未見明顯改變。
[Abstract]:Aim: to investigate the effect of hTERT gene transfection on matrix metalloproteinase 2 and 9 in U2OS cells and to explore the correlation between hTERT gene and telomerase. Methods: RT-PCR and TRAP-ELISA methods were used to detect the transfection effect of hTERT gene and the screening of positive cell clones in U2OS cells, and the effects of hTERT gene transfection on MMP-2,MMP-9 expression and activity were detected by gelatin zymography and RT-PCR. Results: after transfection of hTERT gene into U2OS cells, hTERT mRNA and telomerase activity (p0.05) were found successfully, MMP- (272KD) was decreased (p0.05) and MMP-9 disappeared completely. Conclusion: stable transfection of hTERT into U2OS cell clone completely disappeared the original MMP-9 of the cell secretory enzyme and significantly decreased the original MMP-2 of the enzyme. But MMP-2 did not change obviously in mRNA.
【學位授予單位】:吉林大學
【學位級別】:碩士
【學位授予年份】:2007
【分類號】:R363
本文編號:2281180
[Abstract]:Aim: to investigate the effect of hTERT gene transfection on matrix metalloproteinase 2 and 9 in U2OS cells and to explore the correlation between hTERT gene and telomerase. Methods: RT-PCR and TRAP-ELISA methods were used to detect the transfection effect of hTERT gene and the screening of positive cell clones in U2OS cells, and the effects of hTERT gene transfection on MMP-2,MMP-9 expression and activity were detected by gelatin zymography and RT-PCR. Results: after transfection of hTERT gene into U2OS cells, hTERT mRNA and telomerase activity (p0.05) were found successfully, MMP- (272KD) was decreased (p0.05) and MMP-9 disappeared completely. Conclusion: stable transfection of hTERT into U2OS cell clone completely disappeared the original MMP-9 of the cell secretory enzyme and significantly decreased the original MMP-2 of the enzyme. But MMP-2 did not change obviously in mRNA.
【學位授予單位】:吉林大學
【學位級別】:碩士
【學位授予年份】:2007
【分類號】:R363
【引證文獻】
相關(guān)碩士學位論文 前1條
1 張成斌;胸腺細胞誘導NRK52E細胞轉(zhuǎn)分化及對其遷移能力的影響[D];吉林大學;2012年
,本文編號:2281180
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