本文選題：TNF-α　切入點(diǎn)：P65　出處：《蘭州大學(xué)》2015年碩士論文

【摘要】：目的：本研究探討了急性髓系白血病(Acute myelocytic leukemia, AML)患者和急性髓系白血病細(xì)胞系(HL-60)中TNF-α和P65表達(dá)變化相關(guān)性及意義。材料與方法：收集30例AML患者的骨髓樣本,10例非白血病對(duì)照樣本。通過qRT-PCR檢測(cè)P65和TNF-α的表達(dá),并用Pearson檢驗(yàn)說明樣本中TNF-α和P65表達(dá)的相關(guān)性。受試者工作特征(ROC)曲線觀察在AML患者與非白血病患者TNF-α和P65表達(dá)變化。使用MG132和抗TNF-α抗體分別抑制p65和TNF-α在HL-60細(xì)胞(AML細(xì)胞)中的表達(dá),MTT法檢測(cè)HL-60細(xì)胞的生長(zhǎng)狀況確認(rèn)藥物最佳作用時(shí)間。通過定量qRT-PCR和Western Blot分別檢測(cè)HL-60細(xì)胞中p65和TNF-α mRNA和蛋白的表達(dá)。結(jié)果：1.通過MTT法檢測(cè)結(jié)果顯示,加入抗TNF-α抗體和NF-κB抑制劑MG132培養(yǎng)后的HL-60細(xì)胞生長(zhǎng)受到明顯抑制,生長(zhǎng)曲線低平(P0.05)。2.通過qPT-PCR檢測(cè)骨髓樣本顯示,在急性髓細(xì)胞白血病中P65和TNF-α mRNA相對(duì)表達(dá)量要顯著高于非白血病對(duì)照組；它們可以作為有力的證據(jù)來區(qū)分急性髓細(xì)胞白血病患者的實(shí)驗(yàn)組和非白血病患者的對(duì)照組(P0.05),它們之間存在著顯著的相關(guān)性(R=0.901)。3.通過qRT-PCR檢測(cè)HL-60細(xì)胞株顯示,加入NF-κB抑制劑MG132培養(yǎng)后的細(xì)胞P65和TNF-α mRNA相對(duì)表達(dá)量明顯降低；同樣,加入抗TNF-α抗體培養(yǎng)后,P65和TNF-α mRNA相對(duì)表達(dá)量顯著降低(P0.05)。4.通過Western Blot檢測(cè)HL-60細(xì)胞株顯示,加入NF-κB抑制劑MG132培養(yǎng)后的細(xì)胞P65和TNF-α蛋白相對(duì)表達(dá)量明顯降低；同樣,加入抗TNF-α抗體培養(yǎng)后,P65和TNF-α蛋白相對(duì)表達(dá)量顯著降低(P0.05)。結(jié)論：1.TNF-α和P65在AML中均高表達(dá),并且兩者具有顯著相關(guān)性。2.TNF-α和P65是區(qū)分急性髓細(xì)胞白血病與非白血病未被利用的潛在分子標(biāo)記物。
[Abstract]:Objective: to investigate the correlation and significance of the expression of TNF- 偽 and p65 in acute myelocytic leukemia (AMLA) patients and acute myeloid leukemia cell line (HL-60). Materials and methods: bone marrow samples from 30 patients with AML were collected from 10 cases of non-myeloid leukemia. The expression of p65 and TNF- 偽 was detected by qRT-PCR. The correlation between TNF- 偽 and p65 expression in the sample was demonstrated by Pearson test. The changes of TNF- 偽 and p65 expression in AML patients and non-leukemia patients were observed by using the ROC curve. MG132 and anti-TNF- 偽 antibodies were used to inhibit the expression of TNF- 偽 and TNF- 偽 in HL-60 cells, respectively. The expression of p65 and TNF- 偽 mRNA and protein in HL-60 cells were detected by quantitative qRT-PCR and Western Blot. The growth of HL-60 cells cultured with anti-TNF- 偽 antibody and NF- 魏 B inhibitor MG132 was significantly inhibited, and the growth curve of HL-60 cells was significantly inhibited. The bone marrow samples were detected by qPT-PCR. The relative expression of p65 and TNF- 偽 mRNA in acute myeloid leukemia was significantly higher than that in non-leukemia control group. They can be used as strong evidence to distinguish between the experimental group of acute myeloid leukemia patients and the control group of non-leukemia patients (P0.05). There is a significant correlation between them. The HL-60 cell line was detected by qRT-PCR. The relative expression of p65 and TNF- 偽 mRNA was significantly decreased after the addition of NF- 魏 B inhibitor MG132, and the relative expression of P65 and TNF- 偽 mRNA was significantly decreased after the addition of anti-TNF- 偽 antibody. The expression of P65 and TNF- 偽 mRNA in HL-60 cells was detected by Western Blot assay. The relative expression of p65 and TNF- 偽 protein decreased significantly after the addition of NF- 魏 B inhibitor MG132, and the relative expression of p65 and TNF- 偽 protein decreased significantly after the addition of anti-TNF- 偽 antibody. Conclusion: 1. TNF- 偽 and P65 are highly expressed in AML. TNF- 偽 and p65 are potential molecular markers to distinguish acute myeloid leukemia from non-leukemia.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R733.71

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