线粒体丙酮酸载体与肝细胞肝癌患者预后的相关性研究

发布时间：2018-08-07 17:13

【摘要】：研究背景和目的肝细胞癌是全世界范围内最常见、死亡率最高、恶性程度最高的消化系统恶性肿瘤。临床上最常用和最主要的治疗手段为肝切除术。近年来,随着手术经验、专业知识以及操作技术的不断提高,接受肝切除术的肝细胞癌患者术后的总体生存率有所提高,但术后的无瘤生存率却并没有明显改变,其复发率高达近70%。了解肝细胞癌术后复发的分子机制,寻找可以预测肿瘤患者的预后及复发的肿瘤标记物将会开辟出针对复发性肝细胞癌的新的治疗思路及方法。线粒体是肿瘤发生、发展过程中最重要的介质之一,参与或主导了多种肿瘤细胞特征的转变,包括细胞能量代谢异常、抵抗细胞死亡、组织浸润和转移、促进肿瘤的炎症、基因组不稳定和逃避免疫杀伤。肝细胞含有相对于其他细胞更多的线粒体。因此,线粒体的代谢功能障碍在肝细胞癌的发生、发展中扮演者非常重要的角色。MPC位于线粒体内膜上的丙酮酸转运蛋白,由MPC1和MPC2组成,位于糖酵解和线粒体丙酮酸代谢的交叉点。大多数的肿瘤细胞表现出增强糖酵解和减少氧化磷酸化的特点,因此,MPC应该在肿瘤细胞的代谢改变方面发挥着及其重要的病理生理学作用。但是目前关于MPC对恶性肿瘤发生发展的影响及其机制的研究较少,尤其是在与肝细胞癌方面的研究尚未有报道。本研究通过检测肝细胞癌组织中MPC蛋白及m RNA表达水平,分析MPC在肝细胞癌组织中的表达水平与临床病理学指标的相关性及其对复发和预后的指导价值。以明确MPC对肝细胞癌患者预后的重要意义。希望本研究可以为肝细胞癌的治疗和预后判断及药物的研发提供一条新的思路、观点和依据。研究方法1)通过免疫组织化学法检测85例手术切除的肝细胞癌石蜡标本中癌和癌旁组织的MPC1和MPC2蛋白表达水平,通过Image-Pro Plus软件进行光密度扫描并计算MPC1和MPC2的相对蛋白表达量,分析MPC1和MPC2的蛋白表达水平变化。2)通过Western Blotting检测20例手术切除的肝细胞癌新鲜标本中癌和癌旁组织的MPC1和MPC2蛋白表达水平的变化。3)通过RT-PCR检测20例手术切除的肝细胞癌新鲜标本中癌和癌旁组织的MPC1和MPC2 m RNA表达水平的变化。4)根据MPC1和MPC2相对蛋白表达量的中位数对85例肝细胞肝癌患者进行分组,分析MPC1和MPC2蛋白表达水平和临床病理学指标的相关性。5)根据随访结果和MPC1和MPC2蛋白表达量,Kaplain-Meier生存分析法分析MPC蛋白表达水平与肝细胞癌患者复发及预后的相关性。6)根据随访结果,建立COX回归分析模型,对MPC1和MPC2蛋白表达水平和临床病理学指标进行单因素和多因素分析,寻找肝细胞癌术后复发的独立危险因素。结果1)在手术切除的肝细胞癌石蜡标本中,与癌旁组织相比,癌组织中MPC1和MPC2蛋白表达水平均明显降低。2)在手术切除的肝细胞癌新鲜标本中,与癌旁组织相比,癌组织中MPC1和MPC2蛋白表达水平明显降低。3)在手术切除的肝细胞癌新鲜标本中,与癌旁组织相比,癌组织中MPC1的m RNA表达水平均降低;MPC2 m RNA表达水平则表现为一些标本的癌组织中MPC2的基因表达水平降低,另一些则是癌旁组织中的MPC2基因表达水平降低的无序状态。4)MPC1和MPC2的蛋白表达水平和临床病理学指标均无明显相关性(P0.05)。5)MPC1蛋白表达水平低的肝细胞癌患者的术后复发率升高且总体生存时间缩短;MPC2的蛋白表达水平与复发和总生存期无相关性。6)MPC1蛋白表达水平降低和微血管侵润是肝细胞癌患者术后复发的独立危险因素;单因素和多因素分析显示MPC2和患者术后复发无关。结论在肝细胞癌中MPC1和MPC2的蛋白活性下降或缺失是普遍存在的,且MPC1的蛋白表达水平与患者的术后复发和预后存在明显的相关性,可成为新的判断肝细胞癌患者临床预后、对术后患者进行风险分层的生物标记物。研究结果揭示MPC可能在肝细胞癌糖异生减低和糖酵解增强的代谢异常中发挥重要作用,有望成为一个新的肝细胞癌药物治疗靶点。
[Abstract]:Background and objective hepatocellular carcinoma is the most common, highest mortality and most malignant digestive system malignant tumor in the world. Hepatectomy is the most commonly used and most important treatment in clinic. In recent years, with the experience of surgery, professional knowledge and the continuous improvement of operation techniques, hepatectomy for hepatocarcinoma patients The overall survival rate of the patients was improved, but the postoperative tumor free survival was not significantly changed. The recurrence rate was up to 70%. to understand the molecular mechanism of postoperative recurrence of hepatocellular carcinoma. To find out the prognosis and recurrence of tumor markers that could predict the prognosis and recurrence of tumor patients will open up a new way of treatment for recurrent hepatocellular carcinoma. Mitochondrion is one of the most important mediators in the development of tumor. It participates in or dominates a variety of tumor cell characteristics, including cell energy metabolism abnormality, resistance to cell death, tissue infiltration and metastasis, inflammation of the tumor, genomic instability and escape from epidemic killing. Liver cells contain more than other cells. Mitochondria. Therefore, mitochondrial metabolic dysfunction plays a very important role in the development of hepatocellular carcinoma. The pyruvate transporter, located on the mitochondrial membrane, is composed of MPC1 and MPC2, located at the intersection of glycolysis and mitochondrial pyruvate metabolism. Most of the tumor cells show enhanced glycolysis and reduction. Therefore, MPC should play an important role in the metabolic changes of tumor cells and its important pathophysiological role. However, there are few studies on the effect of MPC on the development of malignant tumor and its mechanism, especially in the study of hepatocellular carcinoma. This study is based on the detection of liver cells. The expression level of MPC protein and m RNA in cancer tissues, the correlation between the expression level of MPC in the hepatocellular carcinoma tissue and the clinicopathological indexes and the guiding value for the recurrence and prognosis. To clarify the significance of MPC to the prognosis of the patients with hepatocellular carcinoma. I hope this study can be used as the treatment and prognosis of the liver cell carcinoma and the development of the drug. A new idea, viewpoint and basis. Method 1) the expression of MPC1 and MPC2 protein in the paraffin tissues of 85 surgically excised hepatocellular carcinoma specimens was detected by immunohistochemistry. The Image-Pro Plus software was used to perform light density scanning and to calculate the relative protein expression of MPC1 and MPC2, and the analysis of MPC1 and MPC2. The change of protein expression level.2) Western Blotting detection of MPC1 and MPC2 protein expression levels in the fresh specimens of hepatocarcinoma in 20 surgically resected hepatocellular carcinoma specimens.3) by RT-PCR detection of the RNA expression level of MPC1 and MPC2 m in the fresh specimens of hepatocarcinoma in 20 cases of surgical resection of hepatocellular carcinoma The median of the relative protein expression of C1 and MPC2 was grouped in 85 patients with hepatocellular carcinoma. The correlation between the expression level of MPC1 and MPC2 protein and the correlation of the clinicopathological indexes was analyzed. According to the follow-up results and the expression of MPC1 and MPC2 protein, the expression of MPC protein and the recurrence and prognosis of the patients with hepatocellular carcinoma were analyzed by Kaplain-Meier survival analysis. Correlation.6) based on the follow-up results, the COX regression analysis model was established to analyze the MPC1 and MPC2 protein expression level and the clinicopathological index by single factor and multifactor analysis to find independent risk factors for the recurrence of hepatocellular carcinoma. Results 1) in the surgical excised hepatocellular carcinoma paraffin specimens, MPC1 and MPC1 were compared with the para cancerous tissue. The expression level of MPC2 protein was significantly reduced by.2). The expression level of MPC1 and MPC2 protein in the cancerous tissue was significantly lower in the fresh specimens of the surgically resected hepatocellular carcinoma than that of the para cancerous tissue. In the fresh specimens of the hepatocarcinoma, the expression level of MPC1 m RNA in the cancerous tissues decreased, and MPC2 m RNA expressed the water. In some specimens, the expression level of MPC2 is reduced in some specimens, and the other is the disordered state of the MPC2 gene expression level in the para cancerous tissue.4). There is no significant correlation between the protein expression level and the clinicopathological index of the MPC1 and MPC2 (P0.05).5) the postoperative recovery of the hepatocellular carcinoma with low level of MPC1 protein expression. Higher hair rate and shorter overall survival time; MPC2 protein expression level and recurrence and total survival time without.6) MPC1 protein expression level and microvascular invasion are independent risk factors for postoperative recurrence of hepatocellular carcinoma; single factor and multiple factor analysis show that MPC2 and patients have no recurrence after operation. Conclusion MPC1 in hepatocellular carcinoma (MPC1) The decrease or loss of protein activity of MPC2 is common, and there is a significant correlation between the protein expression level of MPC1 and the postoperative recurrence and prognosis of the patients. It can be a new clinical prognosis for patients with hepatocellular carcinoma and the risk stratification of the patients after the operation. The results reveal that MPC may be in the sugar isogenesis of hepatocellular carcinoma. It plays an important role in reducing metabolic abnormalities and enhancing glycolysis, and is expected to become a new target for drug therapy of hepatocellular carcinoma.
【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.7

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