本文選題：FasL　切入點(diǎn)：miR-590　出處：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的通過生物信息學(xué)預(yù)測并找出確實(shí)針對Fas L的特異性Micro RNA(miRNA),研究其對舌鱗癌細(xì)胞SCC-3 Fas配體(Fas Lignad,Fas L)的表達(dá)的影響,并最終驗(yàn)證其對舌鱗癌免疫逃逸的影響,從而了解腫瘤的發(fā)生發(fā)展和調(diào)控機(jī)制。方法將合成的miR-590 mimics或miR-590的抑制劑(inhibior)轉(zhuǎn)染至SCC-3細(xì)胞。通過反轉(zhuǎn)錄PCR(reverse transcription-PCR,RT-PCR)和蛋白質(zhì)印跡法(Western Blot)檢測SCC-3中Fas L信使RNA(Messenger RNA,m RNA)和蛋白的表達(dá);將轉(zhuǎn)染后的SCC-3細(xì)胞在DDP(5ug/ml)處理下,在不同時間段通過四甲基偶氮唑藍(lán)(MTT)法繪制細(xì)胞生長曲線,并通過針對Fas L的miR-590 inhibior從反面驗(yàn)證miR-590的作用。結(jié)果將miR-590 mimics轉(zhuǎn)染至舌鱗癌細(xì)胞SCC-3,轉(zhuǎn)染48小時后提取細(xì)胞總RNA,檢測該組與對照組SCC-3細(xì)胞中Fas L m RNA和蛋白的表達(dá)情況后發(fā)現(xiàn),miR-590可以在m RNA和蛋白水平上降低SCC-3細(xì)胞Fas L的表達(dá)。而將miR-590inhibitor轉(zhuǎn)染至SCC-3,檢測Fas L m RNA和蛋白的表達(dá)情況可發(fā)現(xiàn),與對照組相比,表達(dá)miR-590 inhibitor的SCC-3細(xì)胞中Fas L m RNA和蛋白水平顯著升高,增強(qiáng)了Fas L的轉(zhuǎn)錄。SCC-3細(xì)胞轉(zhuǎn)染miR-590后按時間梯度加入DDP檢測可發(fā)現(xiàn),實(shí)驗(yàn)組細(xì)胞存活率低于對照組;轉(zhuǎn)染miR-590 inhibitor反向驗(yàn)證可發(fā)現(xiàn)實(shí)驗(yàn)組細(xì)胞存活率高于對照組。結(jié)論miR-590可以有效抑制SCC-3細(xì)胞Fas L的表達(dá),降低SCC-3的DDP的耐藥性,從而可能降低和減少其對免疫活性細(xì)胞的死亡誘導(dǎo)和殺傷能力,并最終阻斷了癌癥細(xì)胞躲避免疫系統(tǒng)殺傷作用的途徑,為舌鱗癌的生物治療提供了新的靶點(diǎn)。
[Abstract]:Objective to predict and find out the specific Micro RNAs specific to Fas L by bioinformatics, to study its effect on the expression of SCC-3 Fas ligand FAS Lignadfas L in tongue squamous cell carcinoma, and to verify its effect on the immune escape of tongue squamous cell carcinoma.In order to understand the tumorigenesis, development and regulatory mechanisms.Methods miR-590 mimics or miR-590 inhibitor was transfected into SCC-3 cells.The expression of Fas L messenger RNA(Messenger RNAs and proteins in SCC-3 were detected by reverse transcription PCR(reverse transcription-PCRR-RT-PCRand Western blot.The transfected SCC-3 cells were treated with DDPn5ugrml. the growth curves of the transfected SCC-3 cells were plotted by tetramethyl azolium blue at different time intervals.The function of miR-590 is verified by miR-590 inhibior of Fas L.Results miR-590 mimics was transfected into tongue squamous cell SCC-3, and total RNAs were extracted 48 hours after transfection. The expression of Fas L m RNA and protein in SCC-3 cells was detected. It was found that miR-590 could decrease Fas L expression in SCC-3 cells at the level of m RNA and protein.When miR-590inhibitor was transfected into SCC-3, the expression of Fas LM RNA and protein was detected. Compared with the control group, the level of Fas LM RNA and protein in the SCC-3 cells expressing miR-590 inhibitor was significantly higher than that in the control group.After enhanced Fas L transcription. SCC-3 cells were transfected with miR-5 90. The survival rate of the experimental group was lower than that of the control group, and the survival rate of the experimental group was higher than that of the control group.Conclusion miR-590 can effectively inhibit the expression of Fas L in SCC-3 cells and reduce the drug resistance of SCC-3 DDP, which may reduce the death induction and killing ability of SCC-3 cells.Finally, it blocked the way of cancer cells avoiding the killing effect of immune system, and provided a new target for the biotherapy of tongue squamous cell carcinoma.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.86

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本文編號：1695588