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

【摘要】：目的通過生物信息學預測并找出確實針對Fas L的特異性Micro RNA(miRNA),研究其對舌鱗癌細胞SCC-3 Fas配體(Fas Lignad,Fas L)的表達的影響,并最終驗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)展和調控機制。方法將合成的miR-590 mimics或miR-590的抑制劑(inhibior)轉染至SCC-3細胞。通過反轉錄PCR(reverse transcription-PCR,RT-PCR)和蛋白質印跡法(Western Blot)檢測SCC-3中Fas L信使RNA(Messenger RNA,m RNA)和蛋白的表達;將轉染后的SCC-3細胞在DDP(5ug/ml)處理下,在不同時間段通過四甲基偶氮唑藍(MTT)法繪制細胞生長曲線,并通過針對Fas L的miR-590 inhibior從反面驗證miR-590的作用。結果將miR-590 mimics轉染至舌鱗癌細胞SCC-3,轉染48小時后提取細胞總RNA,檢測該組與對照組SCC-3細胞中Fas L m RNA和蛋白的表達情況后發(fā)現(xiàn),miR-590可以在m RNA和蛋白水平上降低SCC-3細胞Fas L的表達。而將miR-590inhibitor轉染至SCC-3,檢測Fas L m RNA和蛋白的表達情況可發(fā)現(xiàn),與對照組相比,表達miR-590 inhibitor的SCC-3細胞中Fas L m RNA和蛋白水平顯著升高,增強了Fas L的轉錄。SCC-3細胞轉染miR-590后按時間梯度加入DDP檢測可發(fā)現(xiàn),實驗組細胞存活率低于對照組;轉染miR-590 inhibitor反向驗證可發(fā)現(xiàn)實驗組細胞存活率高于對照組。結論miR-590可以有效抑制SCC-3細胞Fas L的表達,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胞的死亡誘導和殺傷能力,并最終阻斷了癌癥細胞躲避免疫系統(tǒng)殺傷作用的途徑,為舌鱗癌的生物治療提供了新的靶點。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R739.86

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本文編號：1695588