H3K9me3在腺样囊性癌预后及基因调控中的作用

发布时间：2018-05-01 10:07

本文选题：腺样囊性癌 + 组蛋白H3赖氨酸9三甲基化　；参考：《上海交通大学》2014年博士论文

【摘要】：目的:分析组蛋白H3赖氨酸9三甲基化(H3 lysine 9 trimethylation,H3K9me3)与唾液腺腺样囊性癌(adenoid cystic carcinoma,ACC)患者临床病理特征和预后的关系,并探讨H3K9me3对ACC中相关基因的调控作用及其对ACC生物学行为的影响。方法:免疫组织化学分析66例ACC样本中H3K9me3、EDNRB和CHL1蛋白表达及与临床病理特征、预后的关系。染色质免疫共沉淀结合启动子芯片技术(Ch IP on chip)检测ACC细胞中可能与H3K9me3结合的基因,通过在线工具分析结果。Western Blot方法检测28对肿瘤和瘤旁组织中EDNRB和CHL1蛋白表达,Ch IP-PCR检测ACC组织中H3K9me3与EDNRB和CHL1基因启动子区结合情况。Chaetocin处理细胞后检测EDNRB和CHL1 m RNA表达水平改变。统计分析采用SPSS软件分析。结果:1.H3K9me3表达在ACC中明显高于瘤旁腺体(P0.001),在实体型中明显高于筛状管状型(P=0.002),在发生远处转移者明显高于未发生远处转移者(P=0.001)。H3K9me3高表达的患者总体生存率、无病生存率都比H3K9me3低表达组低。H3K9me3高表达可以作为独立预测ACC总生存率和无病生存率的预测因子。2.SACC-83、SACC-LM中基因启动子区与H3K9me3结合的基因分别为877、1191个,其中两个细胞系均有的基因有551个,这些基因主要参与了嗅觉传导和神经活性配体受体相互作用信号通路以及与G蛋白偶联受体和细胞粘附等。最终筛选出ACC中与H3K9me3关系较为密切的两个基因,分别为EDNRB和CHL1。3.EDNRB和CHL1蛋白在ACC组织中表达低于瘤旁腺体组织。H3K9me3与EDNRB和CHL1启动子区结合有区域特异性。H3K9me3主要结合于EDNRB基因转录起始位点上游-1069~-877bp、-1332~-1111bp和-373~-176bp区域。H3K9me3主要结合于CHL1基因转录起始位点上游-1997~-1803bp和-239~-70bp区域。这种结合可能是这些蛋白表达降低的机制之一,Chaetocin处理SACC-83和SACC-LM可使EDNRB和CHL1 m RNA水平升高。4.T晚期ACC患者EDNRB表达低于T早期患者(P=0.039),CHL1低表达的患者呈现出肿瘤易发生淋巴结转移的趋势(P=0.062)。结论:ACC中H3K9me3的表达明显高于瘤旁腺体中的表达,H3K9me3表达可以作为独立预测ACC总生存率和无病生存率的预测因子。H3K9me3主要结合于EDNRB基因转录起始位点上游-1069~-877bp、-1332~-1111bp和-373~-176bp区域,而在CHL1中主要结合于基因转录起始位点上游-1997~-1803bp和-239~-70bp区域。H3K9me3可能通过调控EDNRB、CHL1的表达在部分肿瘤中促进肿瘤的生长、淋巴结转移。H3K9me3对ACC的组织学亚型、远处转移、预后的影响是通过调节其它基因的功能实现的。
[Abstract]:Objective: to analyze the relationship between histone H3 lysine 9 trimethylation H3 lysine 9 trimethylation (H3K9me3) and the clinicopathological characteristics and prognosis of salivary adenoid cystic carcinoma (ACCs), and to investigate the regulatory role of H3K9me3 in ACC and its effect on the biological behavior of ACC. Methods: the expression of H3K9ME3EDNRB and CHL1 protein and its relationship with clinicopathological features and prognosis in 66 cases of ACC were analyzed by immunohistochemistry. Chromatin immunoprecipitation combined with promoter chip technique was used to detect H3K9me3 binding genes in ACC cells. The expression of EDNRB and CHL1 protein in 28 pairs of tumor tissues and adjacent tissues was detected by online tool analysis. Western Blot method was used to detect the binding of H3K9me3 to EDNRB and CHL1 promoter region in ACC tissue. The expression of EDNRB and CHL1 m RNA were detected after treated with H3K9me3. The statistical analysis was carried out by SPSS software. Results 1. The expression of H3K9me3 in ACC was significantly higher than that in the adjacent gland (P0.001), and in the solid type was significantly higher than that in the sieve tubular type (P0. 002). The overall survival rate of the patients with distant metastasis was significantly higher than that of the patients with high expression of P0. 001. H3K9me3 in patients without distant metastasis. The disease-free survival rate was lower than that in the low expression group of H3K9me3. H3K9me3 expression could be used as an independent predictor of the overall and disease-free survival rates of ACC. 2. The number of genes binding to H3K9me3 in SACC-83nSACC-LM was 877or 1191, respectively. There are 551 genes in both cell lines, which are involved in olfactory conduction, neuroactive ligand receptor interaction signaling pathway, G-protein coupled receptor and cell adhesion. Finally, two genes in ACC which are closely related to H3K9me3 were screened out. The expression of EDNRB, CHL1.3.EDNRB and CHL1 protein in ACC tissue is lower than that in adjacent gland tissue. H3K9me3 binds to EDNRB and CHL1 promoter region. H3K9me3 mainly binds to EDNRB gene transcriptional initiation site upstream -1069n -877bpfU -1332ng-1111bp and -373-176bp region. H3K9me3 mainly binds to EDNRB gene transcription initiation site upstream. H3K9me3 mainly binds to H3K9me3. The region of-1997-1803 BP and-239-70 BP were upstream of CHL1 gene transcription initiation site. This binding may be one of the mechanisms of decreasing the expression of these proteins. Chaetocin treatment with SACC-83 and SACC-LM can increase the level of EDNRB and CHL1 m RNA. 4. The EDNRB expression in late ACC patients is lower than that in early T patients. Conclusion the expression of H3K9me3 in EDNRB is significantly higher than that in adjacent glands. The expression of H3K9me3 can be used as an independent predictor of overall and disease-free survival of ACC. H3K9me3 mainly binds to the upstream region of EDNRB gene transcriptional initiation locus -1069 ~ (-877) BP ~ (-1) 32 ~ (32) ~ (-1111) BP and ~ -3733 ~ (-176) BP. In CHL1, the binding to the upstream of gene transcription initiation locus -1997 ~ 1803bp and -239A ~ 70bp region. H3K9me3 may promote the growth of some tumors by regulating the expression of EDNRBN CHL1. Lymph node metastasis. H3K9me3 has histological subtype and distant metastasis of ACC. The effect of prognosis is achieved by regulating the function of other genes.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R739.8

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