【摘要】：近年來(lái)的研究表明SALL4在許多種腫瘤中高表達(dá)，是候選的原癌基因。然而有關(guān)miRNA與SALL4的調(diào)控關(guān)系的研究報(bào)道較少，SALL4在腫瘤中的表達(dá)調(diào)控機(jī)制還不是很清楚。我們用qRT-PCR技術(shù)分析了miR-497在50例腦膠質(zhì)瘤組織樣品中的表達(dá)情況，結(jié)果表明84%（42/50）的腫瘤組織樣品中miR-497的表達(dá)水平顯著低于對(duì)照組，結(jié)合課題組前期SALL4在同種膠質(zhì)瘤中的表達(dá)水平實(shí)驗(yàn)結(jié)果，并通過(guò)miR-497與SALL4的表達(dá)相關(guān)性分析顯示，miR-497與SALL4具有顯著的負(fù)相關(guān)性。本論文通過(guò)生物信息預(yù)測(cè)發(fā)現(xiàn)SALL4是miR-497的靶基因。應(yīng)用雙熒光素酶報(bào)告系統(tǒng)、qRT-PCR和western-blot技術(shù)發(fā)現(xiàn)，miR-497的過(guò)表達(dá)在mRNA水平和蛋白水平均顯著抑制SALL4的表達(dá)，確認(rèn)SALL4為miR-497的靶基因。通過(guò)MTT增殖曲線、軟瓊脂集落形成實(shí)驗(yàn)，我們發(fā)現(xiàn)miR-497對(duì)于腦膠質(zhì)瘤細(xì)胞的增殖能力具有顯著抑制作用。為了探討miR-497抑制膠質(zhì)瘤增殖的作用機(jī)制，，我們通過(guò)PI-Annexin V雙染及Caspase-3/7活性分析顯示，miR-497可以誘導(dǎo)腦膠質(zhì)瘤細(xì)胞的凋亡。同時(shí)應(yīng)用western blot技術(shù)檢測(cè)FADD的表達(dá)，結(jié)果顯示miR-497過(guò)表達(dá)的腦膠質(zhì)瘤細(xì)胞中，F(xiàn)ADD顯著高表達(dá)。 上述實(shí)驗(yàn)結(jié)果表明，miR-497可以通過(guò)調(diào)控SALL4的表達(dá)來(lái)誘導(dǎo)細(xì)胞凋亡并抑制腦膠質(zhì)瘤細(xì)胞的增殖能力。
[Abstract]:Recent studies have shown that SALL4 is highly expressed in many kinds of tumors and is a candidate proto-oncogene. However, there are few studies on the regulatory relationship between miRNA and SALL4, and the mechanism of SALL4 expression in tumor is not clear. We analyzed the expression of miR-497 in 50 cases of glioma by qRT-PCR technique. The results showed that the expression of miR-497 in 84% (42 / 50) of tumor tissues was significantly lower than that in the control group. According to the experimental results of the expression of SALL4 in the same glioma and the correlation analysis of the expression of miR-497 and SALL4, it was found that there was a significant negative correlation between miR-497 and SALL4. In this paper, we found that SALL4 is the target gene of miR-497 by biological information prediction. Using double luciferase reporting system, qRT-PCR and western-blot techniques, it was found that the overexpression of miR-497 significantly inhibited the expression of SALL4 at both mRNA and protein levels. SALL4 was identified as the target gene of miR-497. Through the MTT proliferation curve and soft Agar colony formation test, we found that miR-497 has a significant inhibitory effect on the proliferation of glioma cells. In order to investigate the mechanism of miR-497 inhibiting glioma proliferation, PI-Annexin V double staining and Caspase-3/7 activity analysis showed that miR-497 could induce apoptosis of glioma cells. At the same time, the expression of FADD was detected by western blot technique. The results showed that FADD was significantly overexpressed in glioma cells with overexpression of miR-497. These results suggest that miR-497 can induce apoptosis and inhibit the proliferation of glioma cells by regulating the expression of SALL4.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.41

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相關(guān)期刊論文 前1條

1 陳忠平,周旺寧;我國(guó)膠質(zhì)瘤診斷治療現(xiàn)狀和努力方向[J];中國(guó)腫瘤;2005年02期

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