HMGB1在非小细胞肺癌及急性肺损伤中的作用机制研究

发布时间：2018-06-21 15:18

本文选题：高迁移率族蛋白 + 非小细胞肺癌　；参考：《山东大学》2012年博士论文

【摘要】：肺癌的发病率和死亡率都持续增高,探讨肺癌的相关细胞因子及发病机制,对减少肺癌发病率有非常重要的作用。研究表明,HMGB1与非小细胞肺癌的发生、发展、转移有非常密切的关系。高迁移率族蛋白B1(high-mobility group box-B1, HMGB1)广泛存在于真核生物细胞内,在许多生物学过程中,HMGB1都有非常重要的作用。HMGB1的表达水平增高可以导致细胞分化、细胞迁移并促进细胞增殖。HMGB1可以促进细胞生长和基因转录,高迁移率族蛋白与肿瘤的许多生物学特性也有非常密切的关系,如肿瘤的发生、浸润、转移等。高迁移率族蛋白的各种生物学活性都是与其相应受体的结合而实现的,晚期糖基化终末产物(RAGE)受体是HMGB1的最主要受体。当HMGB1与RAGE结合时,有两条主要的下游信号转导通路被激活：第一条通路是鸟苷三磷酸酶(Rac)和CDC42途径被激活,产生相应的生物学效应,包括细胞骨架的重塑、细胞运动、细胞迁移和肿瘤的生长、迁移及增殖；第二条通路是丝裂原活化蛋白激酶途径被激活,在这个过程中核因子-κB (NF κB)被活化,产生了大量的细胞因子和趋化因子,促使免疫细胞的成熟,在免疫受体的表达也有非常重要的作用；可能还可以激活另外一条信号转导通路即MAPK, P38, JNK及P42/P44等信号通路,这条通路可以激活MMP—2和—9,二者是系统纤维蛋白溶酶激活级联的下游目标,通过系统纤维蛋白溶酶降解细胞外基质,使肿瘤进一步浸润和转移。目的本研究的目的是通过检测非小细胞肺癌的HMGB1表达水平的变化,HMGB1与肿瘤临床生物学特性的关系,进一步探讨和研究HMGB1在非小细胞肺癌中的作用及发病机制。方法应用逆转录聚合酶链反应(RT-PCR)方法检测HMGB1在非小细胞肺癌中的mRNA的表达；应用蛋白印迹法检测非小细胞肺癌HMGB1蛋白水平的表达；比较肺癌不同分期、不同大小、手术前后HMGB1蛋白水平的变化。结果 1RT-PCR检测人肺癌A549细胞系、支气管上皮细胞系HBE中HMGB1mRNA的表达结果支气管上皮细胞系HBE为对照细胞,根据公式2-average△△CT×100%计算目的基因相对表达量。结果显示肺腺癌A549细胞系HMGB1表达量明显高于支气管上皮细胞系HBE,为其11.450倍。 2人肺癌A549细胞系、支气管上皮细胞系HBE HMGB1蛋白的表达人肺腺癌细胞株A549、支气管上皮细胞系HBE蛋白相对表达量分别为80.44±3.51、6.83±0.75、肺癌细胞株A549HMGB1蛋白表达水平明显高于支气管上皮细胞系HBE,并存在统计学差异(P0.001)。图2显示两株细胞HMGB1蛋白表达的差异。 3肺癌患者与肺良性病患者血清HMGB1蛋白的表达 145例肺癌患者血清HMGB1平均水平为76.1±37.0ng/ml,肺良性病组HMGB1平均水平为39.8±10.8ng/ml),健康志愿者HMGN1平均水平为7.7±6.1ng/ml,肺癌HMGB1表达水平明显高于良性病患者及健康对照组,并且有统计学差异(p0.001)。 4肺癌不同分期血清HMGB1蛋白的表达肺癌Ⅰ,Ⅱ,Ⅲ及Ⅳ期患者血清HMGBl水平分别为30.2±5.9ng/ml,60.9±22.5ng/ml,99.0±23.1ng/ml及133.4±18.9ng/ml。四组间有显著的统计学差异(p0.0001),并且每两组间也有显著的统计学差异。进一步的研究发现,有远处转移的肺癌患者其HMGB1水平为(133.4±18.9ng/ml)明显高于无远处转移的肺癌患者(68.46±31.9ng/ml,p
[Abstract]:The incidence and mortality of lung cancer continue to increase. To explore the cytokine and pathogenesis of lung cancer has a very important role in reducing the incidence of lung cancer. The study shows that HMGB1 is closely related to the occurrence, development and metastasis of non-small cell lung cancer.
High mobility group protein B1 (high-mobility group box-B1, HMGB1) is widely found in eukaryotic cells. In many biological processes, HMGB1 has a very important role in the expression level of.HMGB1, which can lead to cell differentiation. Cell migration and proliferation of cell proliferation and proliferation of.HMGB1 can promote cell growth and gene transcription and high mobility. The family proteins are also closely related to many biological characteristics of tumors, such as tumours, infiltration and metastasis.
All kinds of biological activities of high mobility group proteins are combined with their corresponding receptors, and late glycosylated terminal product (RAGE) receptor is the most important receptor of HMGB1. When HMGB1 and RAGE are combined with RAGE, two major downstream signal transduction pathways are activated: the first pathway is the excitation of guanosine three phosphatase (Rac) and CDC42 pathway Biological effects, including the remodeling of cytoskeleton, cell movement, cell migration and tumor growth, migration and proliferation; the second pathway is activated by mitogen activated protein kinase pathway, in which the nuclear factor kappa B (NF kappa B) is activated, producing a large number of cytokines and chemokines to induce immunization. The maturation of the cell is also very important in the expression of the immune receptor; it may also activate another signal transduction pathway, such as MAPK, P38, JNK and P42/P44, which can activate MMP - 2 and - 9. The two is the downstream target of the system of system fibrinolytic enzyme activation cascade, and is degraded through systematic fibrinolytic enzyme degradation. The extracellular matrix makes the tumor infiltrate and metastases further.
objective
The purpose of this study is to further explore and study the role and pathogenesis of HMGB1 in non-small cell lung cancer by detecting the changes in the expression level of HMGB1 in non-small cell lung cancer, the relationship between HMGB1 and the clinical biological characteristics of the tumor.
Method
The expression of mRNA in non-small cell lung cancer was detected by reverse transcription polymerase chain reaction (RT-PCR), and the expression of HMGB1 protein in non small cell lung cancer was detected by Western blot, and the changes of the level of HMGB1 protein in different stages and sizes of lung cancer before and after operation were compared.
Result
The expression of HMGB1mRNA in human lung cancer A549 cell line and bronchial epithelial cell line HBE was detected by 1RT-PCR.
The bronchial epithelial cell line HBE was a control cell, and the relative expression of the target gene was calculated according to the formula 2-average Delta CT x 100%. The results showed that the expression of HMGB1 in the A549 cell line of lung adenocarcinoma was significantly higher than that of the bronchial epithelial cell line HBE, which was 11.450 times that of the cell line.
Expression of HBE HMGB1 protein in 2 human lung cancer cell line A549 and bronchial epithelial cell line
The relative expression of HBE protein in the human lung adenocarcinoma cell line A549 and the bronchial epithelial cell line was 80.44 + 3.51,6.83 + 0.75 respectively. The expression level of A549HMGB1 protein in lung cancer cell lines was significantly higher than that of the bronchial epithelial cell line HBE, and there were statistical differences (P0.001). Figure 2 showed the difference in the expression of HMGB1 protein in the two cell lines.
Expression of serum HMGB1 protein in 3 patients with lung cancer and pulmonary benign diseases
The average serum HMGB1 level of 145 cases of lung cancer was 76.1 + 37.0ng/ml, the average level of HMGB1 in the lung benign disease group was 39.8 + 10.8ng/ml, the average level of HMGN1 in healthy volunteers was 7.7 + 6.1ng/ml, and the expression level of HMGB1 in lung cancer was significantly higher than that of the benign and healthy controls, and there were statistical differences (p0.001).
Expression of serum HMGB1 protein in 4 different stages of lung cancer
The levels of serum HMGBl in patients with stage I, II, III and IV were 30.2 + 5.9ng/ml, 60.9 + 22.5ng/ml, 99 + 23.1ng/ml and 133.4 + 18.9ng/ml., respectively, and there were significant statistical differences (P0.0001), and there were significant differences between the two groups. Further study showed that the level of HMGB1 in patients with distant metastasis was (133.4 +). 18.9ng/ml) was significantly higher in lung cancer patients without distant metastasis (68.46 + 31.9ng/ml, P)
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R734.2;R563.8

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