基于蛋白质组学的肝细胞癌亚型的分子标志物筛选与临床验证
发布时间：2018-09-03 12:38
【摘要】：目的分析不同临床病理特征的HCC亚型的分子表型的特征,初步探讨其不同生物学特征的分子机制,筛选潜在的分子标志物并验证。1.探讨影响HCC根治性切除术后不同时期复发/转移的临床病理因素;2.比较不同肿瘤数目的HCC亚型的蛋白质组学的特征,初步了解不同肿瘤数目的HCC亚型生物学行为差异的分子基础,寻找潜在的分子标志物并验证;3.比较不同肿瘤大小的HCC亚型的蛋白质组学的特征,初步了解不同肿瘤大小的HCC亚型生物学行为差异的分子基础,寻找潜在的分子标志物并验证;4.探讨HK2表达情况与HCC根治性切除术后复发/转移的关系。方法1.回顾性分析HCC患者根治性切除术后的复发/转移情况,以12项相关的临床病理因素为变量,进行单因素和多因素logistic回归分析,按不同复发/转移时期分析其影响因素。2.iTRAQ标记的定量蛋白质组学的方法,比较单发性和多发性HCC的全体蛋白质组的特征,筛选差异表达蛋白,并应用生物信息学技术分析其分子功能、信号通路等;筛选出不同肿瘤数目的HCC亚型潜在的分子标志物并在m RNA和蛋白水平进行验证。3.iTRAQ标记的定量蛋白质组学的方法,比较微小肝癌、小肝癌、大肝癌、巨大肝癌的全体蛋白质组的特征,筛选差异表达蛋白,并应用生物信息学技术分析其分子功能、信号通路等;筛选不同肿瘤大小的HCC亚型的潜在的分子标志物并在m RNA和蛋白水平进行验证。并对差异蛋白MT1G在HCC中的作用进行初步的分子机制的研究。4.采用Western Blot检测差异表达蛋白HK2在HCC癌组织样本中的表达情况,并与HCC复发/转移的相关临床病理因素进行比较研究。回顾性分析HCC患者根治性切除术后的复发/转移情况,以临床病理因素和HK2表达情况为变量,进行单因素和多因素logistic回归分析,分析其影响因素。结果1.HCC根治性切除术后12个月内复发/转移的独立危险因素为发现方式、术前血清AFP值、肿瘤大小和癌细胞分化程度;术后12~24个月复发/转移的独立危险因素为术前血清AFP值和肿瘤数目;术后24个月复发/转移的独立危险因素为术前血清AFP值、肿瘤大小、肿瘤包膜完整性、癌细胞分化程度。2.在多发性HCC(MC组)和单发性HCC(SC组)的癌组织中,与相应的癌旁组织(MN组和SN组)比较,分别鉴定出107种和330种差异蛋白。在MC组,差异蛋白主要集中在UBC信号通路和NFκB信号通路;而在SC组,差异蛋白主要集中于ERK信号通路和NFκB信号通路。HSD17B13只在MC组下调,而HK2只在SC组上调。3.将癌组织和相应的癌旁组织相比较,在微小肝癌、小肝癌、大肝癌、巨大肝癌分别鉴定出88种、69种、118种和215种差异蛋白。在各个不同肿瘤大小的HCC亚型中,ERK1/2和AKT信号通路的变化在肿瘤发生、发展中均发挥了重要作用。在微小肝癌和巨大肝癌,发现了特有的信号通路的变化。确认了一组与HCC的肿瘤大小显著相关的差异蛋白。MT1G在HCC癌组织中表达降低,与肿瘤大小呈负相关,其过表达可以抑制HCC细胞增殖,促进细胞凋亡。4.HK2表达情况与HCC的发现方式、镜下脉管癌栓、肿瘤大小、肿瘤数目相关。HK2高表达组术后累积生存率显著低于低表达组。HK2为HCC根治性切除术后复发/转移的独立危险因素。结论1.HCC根治性切除术后不同时期复发/转移的影响因素不同。肿瘤数目和肿瘤大小是影响HCC术后不同时期复发/转移的独立危险因素。2.不同肿瘤数目的HCC亚型有着不同的分子学特征和信号通路。筛选并鉴定出HSD17B13和HK2两种差异蛋白,可做为不同肿瘤数目的HCC亚型的潜在的分子标志物。3.不同肿瘤大小的HCC亚型具有不同的分子学特征和信号通路。发现了与HCC的肿瘤大小密切相关的一组差异蛋白,可做为不同肿瘤大小HCC亚型的潜在的分子标志物。差异蛋白MT1G可能通过调节AKT信号通路抑制HCC的增殖。4.HK2表达水平与HCC根治性切除术后的预后密切相关,可作为判断HCC复发/转移和预后的一个有效的分子标志物。
[Abstract]:Objective To analyze the molecular phenotypes of HCC subtypes with different clinical and pathological characteristics and to explore the molecular mechanism of their different biological characteristics and to screen potential molecular markers for identification. 3. Comparing the proteomic characteristics of HCC subtypes with different tumor sizes, we can preliminarily understand the molecular basis of the differences in biological behavior of HCC subtypes with different tumor sizes and find out the potential molecular markers. To explore the relationship between the expression of HK2 and the recurrence/metastasis of HCC patients after radical resection. Methods 1. The recurrence/metastasis of HCC patients after radical resection was retrospectively analyzed. 12 clinicopathological factors were taken as variables, and univariate and multivariate logistic regression analysis was performed. 2. Quantitative proteomics with iTRAQ markers was used to compare the proteomic characteristics of single and multiple HCCs, screen differentially expressed proteins, analyze their molecular functions and signal pathways by bioinformatics techniques, and screen out potential molecular markers of HCC subtypes with different tumor numbers in M RNA and m RNA. 3. Quantitative proteomics with iTRAQ markers was used to compare the proteomic characteristics of small, small, large, and giant hepatocellular carcinomas, screen differentially expressed proteins, analyze their molecular functions and signal pathways by bioinformatics techniques, and screen potential subtypes of HCC with different tumor sizes. The expression of differentially expressed protein HK2 in HCC tissues was detected by Western Blot and compared with the clinicopathological factors associated with recurrence/metastasis of HCC. The role of differentially expressed protein MT1G in HCC was studied preliminarily. The recurrence/metastasis of CC patients after radical resection were analyzed by univariate and multivariate logistic regression analysis with clinicopathological factors and HK2 expression as variables. Results 1. The independent risk factors of recurrence/metastasis within 12 months after radical resection of HCC were found by preoperative serum AFP, tumor size and tumor size. The independent risk factors for recurrence/metastasis were preoperative AFP level and tumor number at 12-24 months after surgery. The independent risk factors for recurrence/metastasis at 24 months after surgery were preoperative AFP level, tumor size, tumor envelope integrity, and differentiation of cancer cells. Compared with the corresponding adjacent tissues (MN group and SN group), 107 and 330 differentially expressed proteins were identified. In MC group, the differentially expressed proteins were mainly concentrated in UBC and NF-kappa B signaling pathways, while in SC group, the differentially expressed proteins were mainly concentrated in ERK and NF-kappa B signaling pathways. Compared with the corresponding adjacent tissues, 88, 69, 118 and 215 differential proteins were identified in small, small, large, and giant hepatocellular carcinomas, respectively. The expression of MT1G in HCC tissues was negatively correlated with tumor size. The overexpression of MT1G could inhibit the proliferation of HCC cells and promote apoptosis. 4. The expression of HK2 was associated with the pattern of HCC detection, vascular tumor thrombus and tumor size under microscope. The cumulative survival rate of the patients with high expression of HK2 was significantly lower than that of the patients with low expression of HK2. HK2 was an independent risk factor for recurrence/metastasis after HCC radical resection. Conclusion 1. The factors influencing recurrence/metastasis were different at different stages after HCC radical resection. Independent risk factors. 2. HCC subtypes with different tumor numbers have different molecular characteristics and signaling pathways. Two different proteins, HSD17B1 3 and HK2, were screened and identified as potential molecular markers of HCC subtypes with different tumor numbers. 3. HCC subtypes with different tumor sizes have different molecular characteristics and signaling pathways. Differential protein MT1G may inhibit the proliferation of HCC by regulating AKT signaling pathway. 4. The expression level of HK2 is closely related to the prognosis of HCC after radical resection. It can be used as a predictor of recurrence/metastasis and prognosis of HCC. An effective molecular marker.
【学位授予单位】：福建医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7
[Abstract]:Objective To analyze the molecular phenotypes of HCC subtypes with different clinical and pathological characteristics and to explore the molecular mechanism of their different biological characteristics and to screen potential molecular markers for identification. 3. Comparing the proteomic characteristics of HCC subtypes with different tumor sizes, we can preliminarily understand the molecular basis of the differences in biological behavior of HCC subtypes with different tumor sizes and find out the potential molecular markers. To explore the relationship between the expression of HK2 and the recurrence/metastasis of HCC patients after radical resection. Methods 1. The recurrence/metastasis of HCC patients after radical resection was retrospectively analyzed. 12 clinicopathological factors were taken as variables, and univariate and multivariate logistic regression analysis was performed. 2. Quantitative proteomics with iTRAQ markers was used to compare the proteomic characteristics of single and multiple HCCs, screen differentially expressed proteins, analyze their molecular functions and signal pathways by bioinformatics techniques, and screen out potential molecular markers of HCC subtypes with different tumor numbers in M RNA and m RNA. 3. Quantitative proteomics with iTRAQ markers was used to compare the proteomic characteristics of small, small, large, and giant hepatocellular carcinomas, screen differentially expressed proteins, analyze their molecular functions and signal pathways by bioinformatics techniques, and screen potential subtypes of HCC with different tumor sizes. The expression of differentially expressed protein HK2 in HCC tissues was detected by Western Blot and compared with the clinicopathological factors associated with recurrence/metastasis of HCC. The role of differentially expressed protein MT1G in HCC was studied preliminarily. The recurrence/metastasis of CC patients after radical resection were analyzed by univariate and multivariate logistic regression analysis with clinicopathological factors and HK2 expression as variables. Results 1. The independent risk factors of recurrence/metastasis within 12 months after radical resection of HCC were found by preoperative serum AFP, tumor size and tumor size. The independent risk factors for recurrence/metastasis were preoperative AFP level and tumor number at 12-24 months after surgery. The independent risk factors for recurrence/metastasis at 24 months after surgery were preoperative AFP level, tumor size, tumor envelope integrity, and differentiation of cancer cells. Compared with the corresponding adjacent tissues (MN group and SN group), 107 and 330 differentially expressed proteins were identified. In MC group, the differentially expressed proteins were mainly concentrated in UBC and NF-kappa B signaling pathways, while in SC group, the differentially expressed proteins were mainly concentrated in ERK and NF-kappa B signaling pathways. Compared with the corresponding adjacent tissues, 88, 69, 118 and 215 differential proteins were identified in small, small, large, and giant hepatocellular carcinomas, respectively. The expression of MT1G in HCC tissues was negatively correlated with tumor size. The overexpression of MT1G could inhibit the proliferation of HCC cells and promote apoptosis. 4. The expression of HK2 was associated with the pattern of HCC detection, vascular tumor thrombus and tumor size under microscope. The cumulative survival rate of the patients with high expression of HK2 was significantly lower than that of the patients with low expression of HK2. HK2 was an independent risk factor for recurrence/metastasis after HCC radical resection. Conclusion 1. The factors influencing recurrence/metastasis were different at different stages after HCC radical resection. Independent risk factors. 2. HCC subtypes with different tumor numbers have different molecular characteristics and signaling pathways. Two different proteins, HSD17B1 3 and HK2, were screened and identified as potential molecular markers of HCC subtypes with different tumor numbers. 3. HCC subtypes with different tumor sizes have different molecular characteristics and signaling pathways. Differential protein MT1G may inhibit the proliferation of HCC by regulating AKT signaling pathway. 4. The expression level of HK2 is closely related to the prognosis of HCC after radical resection. It can be used as a predictor of recurrence/metastasis and prognosis of HCC. An effective molecular marker.
【学位授予单位】：福建医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7
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