基于蛋白質(zhì)組學(xué)的肝細(xì)胞癌亞型的分子標(biāo)志物篩選與臨床驗證
發(fā)布時間：2018-09-03 12:38
【摘要】：目的分析不同臨床病理特征的HCC亞型的分子表型的特征,初步探討其不同生物學(xué)特征的分子機制,篩選潛在的分子標(biāo)志物并驗證。1.探討影響HCC根治性切除術(shù)后不同時期復(fù)發(fā)/轉(zhuǎn)移的臨床病理因素;2.比較不同腫瘤數(shù)目的HCC亞型的蛋白質(zhì)組學(xué)的特征,初步了解不同腫瘤數(shù)目的HCC亞型生物學(xué)行為差異的分子基礎(chǔ),尋找潛在的分子標(biāo)志物并驗證;3.比較不同腫瘤大小的HCC亞型的蛋白質(zhì)組學(xué)的特征,初步了解不同腫瘤大小的HCC亞型生物學(xué)行為差異的分子基礎(chǔ),尋找潛在的分子標(biāo)志物并驗證;4.探討HK2表達(dá)情況與HCC根治性切除術(shù)后復(fù)發(fā)/轉(zhuǎn)移的關(guān)系。方法1.回顧性分析HCC患者根治性切除術(shù)后的復(fù)發(fā)/轉(zhuǎn)移情況,以12項相關(guān)的臨床病理因素為變量,進(jìn)行單因素和多因素logistic回歸分析,按不同復(fù)發(fā)/轉(zhuǎn)移時期分析其影響因素。2.iTRAQ標(biāo)記的定量蛋白質(zhì)組學(xué)的方法,比較單發(fā)性和多發(fā)性HCC的全體蛋白質(zhì)組的特征,篩選差異表達(dá)蛋白,并應(yīng)用生物信息學(xué)技術(shù)分析其分子功能、信號通路等;篩選出不同腫瘤數(shù)目的HCC亞型潛在的分子標(biāo)志物并在m RNA和蛋白水平進(jìn)行驗證。3.iTRAQ標(biāo)記的定量蛋白質(zhì)組學(xué)的方法,比較微小肝癌、小肝癌、大肝癌、巨大肝癌的全體蛋白質(zhì)組的特征,篩選差異表達(dá)蛋白,并應(yīng)用生物信息學(xué)技術(shù)分析其分子功能、信號通路等;篩選不同腫瘤大小的HCC亞型的潛在的分子標(biāo)志物并在m RNA和蛋白水平進(jìn)行驗證。并對差異蛋白MT1G在HCC中的作用進(jìn)行初步的分子機制的研究。4.采用Western Blot檢測差異表達(dá)蛋白HK2在HCC癌組織樣本中的表達(dá)情況,并與HCC復(fù)發(fā)/轉(zhuǎn)移的相關(guān)臨床病理因素進(jìn)行比較研究。回顧性分析HCC患者根治性切除術(shù)后的復(fù)發(fā)/轉(zhuǎn)移情況,以臨床病理因素和HK2表達(dá)情況為變量,進(jìn)行單因素和多因素logistic回歸分析,分析其影響因素。結(jié)果1.HCC根治性切除術(shù)后12個月內(nèi)復(fù)發(fā)/轉(zhuǎn)移的獨立危險因素為發(fā)現(xiàn)方式、術(shù)前血清AFP值、腫瘤大小和癌細(xì)胞分化程度;術(shù)后12~24個月復(fù)發(fā)/轉(zhuǎn)移的獨立危險因素為術(shù)前血清AFP值和腫瘤數(shù)目;術(shù)后24個月復(fù)發(fā)/轉(zhuǎn)移的獨立危險因素為術(shù)前血清AFP值、腫瘤大小、腫瘤包膜完整性、癌細(xì)胞分化程度。2.在多發(fā)性HCC(MC組)和單發(fā)性HCC(SC組)的癌組織中,與相應(yīng)的癌旁組織(MN組和SN組)比較,分別鑒定出107種和330種差異蛋白。在MC組,差異蛋白主要集中在UBC信號通路和NFκB信號通路;而在SC組,差異蛋白主要集中于ERK信號通路和NFκB信號通路。HSD17B13只在MC組下調(diào),而HK2只在SC組上調(diào)。3.將癌組織和相應(yīng)的癌旁組織相比較,在微小肝癌、小肝癌、大肝癌、巨大肝癌分別鑒定出88種、69種、118種和215種差異蛋白。在各個不同腫瘤大小的HCC亞型中,ERK1/2和AKT信號通路的變化在腫瘤發(fā)生、發(fā)展中均發(fā)揮了重要作用。在微小肝癌和巨大肝癌,發(fā)現(xiàn)了特有的信號通路的變化。確認(rèn)了一組與HCC的腫瘤大小顯著相關(guān)的差異蛋白。MT1G在HCC癌組織中表達(dá)降低,與腫瘤大小呈負(fù)相關(guān),其過表達(dá)可以抑制HCC細(xì)胞增殖,促進(jìn)細(xì)胞凋亡。4.HK2表達(dá)情況與HCC的發(fā)現(xiàn)方式、鏡下脈管癌栓、腫瘤大小、腫瘤數(shù)目相關(guān)。HK2高表達(dá)組術(shù)后累積生存率顯著低于低表達(dá)組。HK2為HCC根治性切除術(shù)后復(fù)發(fā)/轉(zhuǎn)移的獨立危險因素。結(jié)論1.HCC根治性切除術(shù)后不同時期復(fù)發(fā)/轉(zhuǎn)移的影響因素不同。腫瘤數(shù)目和腫瘤大小是影響HCC術(shù)后不同時期復(fù)發(fā)/轉(zhuǎn)移的獨立危險因素。2.不同腫瘤數(shù)目的HCC亞型有著不同的分子學(xué)特征和信號通路。篩選并鑒定出HSD17B13和HK2兩種差異蛋白,可做為不同腫瘤數(shù)目的HCC亞型的潛在的分子標(biāo)志物。3.不同腫瘤大小的HCC亞型具有不同的分子學(xué)特征和信號通路。發(fā)現(xiàn)了與HCC的腫瘤大小密切相關(guān)的一組差異蛋白,可做為不同腫瘤大小HCC亞型的潛在的分子標(biāo)志物。差異蛋白MT1G可能通過調(diào)節(jié)AKT信號通路抑制HCC的增殖。4.HK2表達(dá)水平與HCC根治性切除術(shù)后的預(yù)后密切相關(guān),可作為判斷HCC復(fù)發(fā)/轉(zhuǎn)移和預(yù)后的一個有效的分子標(biāo)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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R735.7
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