PDCD4基因论文：PDCD4基因急性髓性白血病TGF-β1RT-PCRELISAWesternblotting

PDCD4基因论文：PDCD4基因急性髓性白血病 TGF-β1 RT-PCR ELISA Western blotting【中文摘要】白血病是一类源于造血干细胞的恶性克隆性疾病,白血病细胞在骨髓中大量增生,从而使正常造血功能受到抑制,同时白血病细胞又可浸润其他器官和组织,影响其它组织器官的功能。

虽然目前白血病的发病机制并不十分清楚,但是随着现代实验技术的进步,人类对其发病机制有了进一步的认识和发现,其发生机制可能与基因突变、病毒感染、环境因素、及其他血液系统疾病等有关。

而目前最新的研究表明,在白血病的发生发展中,细胞的无限增生及其调控异常起了重要作用。

而生物体维持自身稳定的重要条件就是细胞增生和细胞凋亡间的平衡状态,如果细胞出现凋亡受阻或增殖失控均有可能导致肿瘤形成。

程序性细胞凋亡因子4(programmed cell death 4, PDCD4)基因是因可以在凋亡中出现上调而得名,但是目前它的确切功能尚不完全清楚。

但是现在越来越多的证据表明,此基因不仅可以影响转录、翻译过程以及可以调节不同的信号转导途径,而且其在抑制肿瘤形成方面可能也起了一定的作用。

TGF-β1是一种常见的多效性细胞因子,它不仅能有效刺激成纤维细胞的生长,修复创伤,而且还能调节造血,并可以抑制肿瘤细胞的增殖。

因此,本课题重点研究PDCD4和TGF-β1在急性髓性白血病骨髓中的表达及意义,初步探讨其在白血病中的作用及可能的机制。

本研究所使用的标本为2009年8月-2009年12月在山东大学齐鲁医院血液科住院患者,其中缺铁性贫血(对照组)20例,男6例,女14例,27～70岁,平均39岁；急性髓系白血病初诊患者(AML组)30例,男18例,女12例,18～73岁,平均46岁；完全缓解组患者(AML-CR组)30例,男16例,女14例,16～67岁,平均41岁。

患者诊断均经临床、血常规、骨髓象及组织化学染色、免疫分型、染色体检查确诊。

采用RT-PCR、ELISA和Western blotting 三种方法分别从基因水平和蛋白水平检测其骨髓单个核细胞中PDCD4、TGF-β1的表达情况,并分析相互间的相关性。

采用SPSS13.0软件对数据进行统计学分析。

p0.05)。

ELISA结果显示,AML组TGF-p1的蛋白表达量明显低于对照组和AML-CR组(P0.05),PDCD4、TGF-p1的表达呈正相关关系(p<0.05)PDCD4mRNA水平和蛋白水平初诊AML中表达明显减低,提示对于初诊AML患者骨髓单个核细胞中凋亡受阻,可能参与了白血病的发生发展。

AML-CR组mRNA表达明显高于AML组,但是没有统计学意义,可能与本研究的样本量少有关,扩大AML-CR组和AML组患者的样本量有可能会发现AML-CR组和AML组之间的差异。

TGF-β1mRNA水平和蛋白水平初诊AML中表达也出现了明显减低,表明初诊AML组患者骨髓中TGF-β1生成减少,使白血病细胞失去了增殖抑制而大量增长,加之缺乏TGF-β1对白血病细胞成熟分化的诱导作用,加速了病情的恶化。

因此检测AML患者的可溶性TGF-β1水平可作为病情诊断和预后判断的指标之一。

初诊AML患者达到完全缓解后,TGF-β1mRNA的表达量虽明显高于AML组,但是没有统计学意义,而TGF-β1蛋白的表达量明显高于AML组,差异有统计学意义,可能是机体多种细胞都可以分泌TGF-p1,也可能是本研究的样本量太少,因此TGF-β1是否可作为完全缓解后的预测指标尚需进一步研究。

研究中发现,PDCD4、TGF-β1的表达呈正相关关系,提示TGF-p1可能是通过调节PDCD4的表达促进白血病细胞凋亡【英文摘要】[Background and ]Leukemia is a malignant clonal disease derived from hematopoietic stem cells, and leukemia cells in the bone marrow and other tissues accumulated a large number of proliferation, and invased of other organs and tissues, so that normal hematopoiesis was inhibited. The mechanism may be related with environmental factors, viral infection, gene mutation, and other blood diseases and so on. Although the pathogenesis of leukemia is not clear, but with the progress of modern experimental methods, the people have a novel knowledge and discovery gradually. The latest study found that cell proliferation and regulation of infinite abnormality played an important role in the development of leukemia. Cell proliferation and apoptosis to maintain the balance of the organism is an important factor, if the blockage of apoptosis or uncontrolled proliferation may result in tumor formation.PDCD4 gene is regulated by apoptosis, but its exact function is unclear. Now more and more evidence showed that the gene can affect the transcription, translation and regulation of different signal transduction pathways and may also play arole in tumor suppression. TGF-β1 is a common pleiotropic cytokine, which can effectively stimulate the growth of fibroblast cells to repair wounds, but also regulate blood and inhibit tumor proliferation. Therefore, this project focuses on the PDCD4 and TGF-(β1 in acute myeloid leukemia and significance of the expression in bone marrow, and explores its role in leukemia and its possible mechanism.[Methods] The sample was collected in Department of Hematology, Qilu Hospital on 2009.8-2009.12. The sample included iron deficiency anemia (control group) 20 cases,6 males and 14 females,27 to 70 years old, an average of 39 years; patients with newly diagnosed acute myeloid leukemia (AML) 30,18 males and 12 females, 18 to 73 years, mean 46 years; complete remission patients (AML-CR) 30,16 males patients,14 females,16 to 67 years, with an average of 41 years. Patients were diagnosed by clinically, blood, bone marrow and tissue staining, immunophenotyping, chromosome diagnosis. RT-PCR, ELISA and Western blotting were three methods of gene and protein levels detected in the bone marrow mononuclear cells PDCD4, TGF-β1 expression, and analysis of the correlation between each other. SPSS13.0 software was used for statistical analysis. p0.05).The results of ELISA showed that the level of TGF-β1 in newly diagnosed AML group wassignificantly lower than the other two groups(P <0.05). The results of Western blotting indicated that expression of PDCD4 protein in AML group was significantly lower than the control group and the AML-CR group(P<0.05).PDCD4,TGF-β1expression had positive correlation(p<0.05).[Conclusion]PDCD4 mRNA and protein levels were significantly lower in newly diagnosed AML, suggesting that patients with newly diagnosed AML in bone marrow cells blocked apoptosis might be involved in the development of leukemia. mRNA expression in AML-CR group was significantly higher than AML group, but not statistically significant, the sample may be less in this study,and expand the sample of the AML-CR group and AML patients may be found in AML-CR group and the AML the difference between the groups. TGF-β1mRNA and protein expression levels in newly diagnosed AML have shown a significant reduction, indicating that the bone marrow of newly diagnosed AML patients decreased production of TGF-β1, the proliferation of leukemia cells but lost a lot of growth, and the lack of TGF-β1 on the leukemia cell maturation induced differentiation and accelerated the deterioration of the condition. Therefore, detection of AML patients with levels of TGF-β1 can be used as diagnosis and prognosis of the disease one of the indicators. Newly diagnosedAML patients achieved complete remission, TGF-β1 mRNA expression, although significantly higher than that of AML group, but not statistically significant, while the expression of TGF-β1 was significantly higher than the AML group, the difference was statistically significant, may be the body cells can secrete a variety of TGF-β1, may be too small sample size of this study, whether TGF-β1 as a predictor of complete remission still need further study. Study found, PDCD4, TGF-β1 expression was positively correlated, suggesting that TGF-β1 may regulate PDCD4 expression by promoting apoptosis of leukemia cells.【关键词】PDCD4基因急性髓性白血病 TGF-β1 RT-PCR ELISA Western blotting【采买全文】1.3.9.9.38.8.4.8 1.3.8.1.13.7.2.1同时提供论文写作定制和论文发表服务.保过包发.【说明】本文仅为中国学术文献总库合作提供，无涉版权。