Rab27A、Rab27B与TP53在胰腺癌中的作用及部分机制研究
发布时间：2019-02-24 12:11
【摘要】：胰腺癌是消化系统常见的恶性肿瘤之一,其恶性程度高、早期诊断率低、进展迅速、预后极差。胰腺癌已经成为国内外医疗界需要共同面对的一个重大的诊断和治疗难题,因此加强胰腺癌发生机制研究,以发现新的诊断标记物与探索新的治疗策略,具有十分重要的意义。生物体内细胞正常的运转,需要让合适的分子在合适的时间到达合理的靶位置,这一重要作用主要依赖囊泡运输完成。其功能障碍会导致多种细胞器发生缺陷,细胞功能紊乱。近年来的研究显示囊泡运输异常与肿瘤的发生发展相关。在与蛋白质囊泡运输有关的效应分子中,Rab蛋白是Ras相关的小GTP酶超家族的成员,这些蛋白有着特定的细胞亚定位,并在细胞的分泌、胞吞、信号转导和发育等过程发挥着重要的作用。有研究表明,Rab27家族中Rab27A及Rab27B与肿瘤的发生及转移有密切的关系,有学者发现Rab27A与人乳腺癌细胞的侵袭和转移密切相关;Rab27B可调节雌激素受体阳性的乳腺癌细胞的侵袭性生长和转移;Rab27B和Rab27A与肝癌的进展相关,可能是肝癌患者一个有意义的预后指标。检索文献关于Rab27B和Rab27A与胰腺癌之间的关系尚未见诸报道。我们通过将186例胰腺癌组织,54例癌旁周围组织和25例胰腺良性疾病组织制成组织芯片,以免疫组化技术分析Rab27A和TP53在胰腺组织中的表达及临床意义。我们发现Rab27A与TP53在胰腺癌组织中较癌旁组织及胰腺良性疾病组织明显增高。Rab27A和TP53的高表达均与胰腺癌的肿瘤分期及远处转移密切相关。Rab27A的高表达还与血管浸润有关。在Cox单因素和多因素分析中,我们发现Rab27A与TP53的阳性表达与胰腺癌患者不良预后有密切关系,差异具有统计学意义。虽然研究数据不支持TP53可通过Rab27A调控胰腺癌的胞外分泌功能,但不能排除二者之间存在较微弱的相关作用的可能性。随后的研究中,采用免疫组织化学方法,组织芯片技术,对186例胰腺癌组织,51例对应的癌旁周围组织和23例胰腺良性病变组织标本中Rab27B及TP53的表达进行检测,结果表明,Rab27B的高表达可作为胰腺癌的独立的预后因子。通过Cox单因素和多因素分析,我们发现Rab27B的表达水平增高是胰腺癌不良预后的重要的危险因素,胰腺癌中Rab27B和TP53表达水平的增高还与血管浸润、远处转移有密切关系,同时检测到Rab27B与TP53蛋白的表达之间有着相关性,而TP53与Rab27B之间有着怎样的作用关系还有待于进一步研究。我们还分别构建了针对Rab27A及Rab27B基因的siRNA,通过脂质体的方法对胰腺癌细胞进行转染,在细胞水平上,分别用CCK-8、Transwell小室及Western Blot等方法检测转染前后对细胞生物学特性的影响。结果显示,干扰Rab27A及Rab27B基因的表达可抑制胰腺癌的增殖,减弱其侵袭能力,并可诱导细胞凋亡,对胰腺癌的增殖转移行为起到反向调控的作用。综上所述,本研究首次报道了Rab27A及TP53在胰腺癌组织中表达较癌旁组织及胰腺良性疾病组织高,且高表达的Rab27A与胰腺癌患者的不良预后之间有着密切关系。Rab27B在胰腺癌中的表达虽与癌旁组织及胰腺良性疾病组织中的表达无明显统计学差异,但在Rab27B高表达的胰腺癌患者,其五年生存率有明显的降低。我们没有检测到Rab27A与TP53在表达上明显的相关,但Rab27B和TP53的表达有显著的相关性,具体的作用方式还有待进一步研究。以siRNA技术干扰Rab27A及Rab27B均可明显降低胰腺癌细胞的增殖、侵袭、迁移能力,诱导凋亡,由此为下一步构建新型靶向治疗药物提供了理论依据和实验基础。
[Abstract]:Pancreatic cancer is one of the most common malignant tumors in the digestive system. Its malignant degree is high, the early diagnosis rate is low, the progress is rapid, and the prognosis is very poor. Pancreatic cancer has become a major diagnosis and treatment problem to be faced by the medical community at home and abroad, so it is of great significance to study the mechanism of pancreatic cancer in order to find new diagnostic markers and explore new therapeutic strategies. The normal operation of the cells in the organism requires the appropriate molecules to reach a reasonable target position at the right time, which is mainly dependent on the completion of the vesicle transport. The dysfunction can lead to the defects of various organelles and the disorder of cell function. In recent years, the study of vesicle transport is related to the development of the tumor. In effector molecules associated with the transport of protein vesicles, the Rab protein is a member of the Ras-associated small GTP-enzyme superfamily, which has specific subcellular localization and plays an important role in the process of cell secretion, endocytosis, signal transduction and development. It is shown that Rab27A and Rab27B in the Rab27 family have a close relationship with the occurrence and metastasis of the tumor, and some scholars have found that Rab27A is closely related to the invasion and metastasis of human breast cancer cells. Rab27B can regulate the invasive growth and metastasis of breast cancer cells that are positive for estrogen receptor. Rab27B and Rab27A are related to the progression of liver cancer, and may be a meaningful prognostic indicator for patients with liver cancer. The literature on the relationship between Rab27B and Rab27A and pancreatic cancer has not been reported. The expression of Rab27A and TP53 in pancreatic tissue and its clinical significance were analyzed by immunohistochemistry. We found that Rab27A and TP53 increased significantly in the tissues of the pancreatic cancer and the tissues of the benign pancreatic diseases in the pancreas. The high expression of Rab27A and TP53 is closely related to the staging and distant metastasis of pancreatic cancer. The high expression of Rab27A is also associated with vascular invasion. In Cox single and multi-factor analysis, we found that the positive expression of Rab27A and TP53 is closely related to the poor prognosis of patients with pancreatic cancer, and the difference is of statistical significance. Although the study data does not support the ability of TP53 to regulate the extracellular secretion of pancreatic cancer by Rab27A, it is not possible to exclude the possibility of a weak correlation between the two. In the following study, the expression of Rab27B and TP53 in the specimens of 186 pancreatic cancer tissues, 51 corresponding adjacent tissues and 23 cases of benign lesions of the pancreas was detected by immunohistochemical method and tissue chip technique. The results showed that, The high expression of Rab27B can be used as an independent prognostic factor for pancreatic cancer. Through the Cox and multi-factor analysis, we found that the increase of the expression level of Rab27B is an important risk factor for the poor prognosis of the pancreatic cancer. The level of the expression of Rab27B and TP53 in the pancreatic cancer is also closely related to the vascular invasion and distant metastasis. The relationship between the expression of Rab27B and TP53 protein is also detected, and the relationship between TP53 and Rab27B remains to be further studied. In addition, the siRNA of Rab27A and Rab27B gene was constructed, and the cells of pancreatic cancer were transfected with the method of liposome, and the effects of the transfection and before and after transfection on the biological characteristics of the cells were detected by the methods of CCK-8, Transwell and Western Blot respectively. The results showed that the expression of Rab27A and Rab27B gene could inhibit the proliferation of pancreatic cancer, reduce its invasion ability and induce cell apoptosis. To sum up, this study first reported the high expression of Rab27A and TP53 in the tissues of pancreatic cancer and the high tissue of the pancreatic benign diseases, and the high expression of Rab27A was closely related to the poor prognosis of the patients with pancreatic cancer. The expression of Rab27B in pancreatic cancer has no significant difference with the expression of the adjacent tissue and the tissue of the pancreatic benign disease, but the five-year survival rate of Rab27B is significantly lower in patients with pancreatic cancer with high expression of Rab27B. We did not detect that Rab27A and TP53 are significantly related to the expression, but the expression of Rab27B and TP53 has a significant correlation, and the specific mode of action is to be further studied. The interference of Rab27A and Rab27B with siRNA technology can significantly reduce the proliferation, invasion, migration and apoptosis of pancreatic cancer cells, thus providing a theoretical basis and experimental basis for the next step to construct a novel targeted therapeutic agent.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R735.9
本文编号：2429539
[Abstract]:Pancreatic cancer is one of the most common malignant tumors in the digestive system. Its malignant degree is high, the early diagnosis rate is low, the progress is rapid, and the prognosis is very poor. Pancreatic cancer has become a major diagnosis and treatment problem to be faced by the medical community at home and abroad, so it is of great significance to study the mechanism of pancreatic cancer in order to find new diagnostic markers and explore new therapeutic strategies. The normal operation of the cells in the organism requires the appropriate molecules to reach a reasonable target position at the right time, which is mainly dependent on the completion of the vesicle transport. The dysfunction can lead to the defects of various organelles and the disorder of cell function. In recent years, the study of vesicle transport is related to the development of the tumor. In effector molecules associated with the transport of protein vesicles, the Rab protein is a member of the Ras-associated small GTP-enzyme superfamily, which has specific subcellular localization and plays an important role in the process of cell secretion, endocytosis, signal transduction and development. It is shown that Rab27A and Rab27B in the Rab27 family have a close relationship with the occurrence and metastasis of the tumor, and some scholars have found that Rab27A is closely related to the invasion and metastasis of human breast cancer cells. Rab27B can regulate the invasive growth and metastasis of breast cancer cells that are positive for estrogen receptor. Rab27B and Rab27A are related to the progression of liver cancer, and may be a meaningful prognostic indicator for patients with liver cancer. The literature on the relationship between Rab27B and Rab27A and pancreatic cancer has not been reported. The expression of Rab27A and TP53 in pancreatic tissue and its clinical significance were analyzed by immunohistochemistry. We found that Rab27A and TP53 increased significantly in the tissues of the pancreatic cancer and the tissues of the benign pancreatic diseases in the pancreas. The high expression of Rab27A and TP53 is closely related to the staging and distant metastasis of pancreatic cancer. The high expression of Rab27A is also associated with vascular invasion. In Cox single and multi-factor analysis, we found that the positive expression of Rab27A and TP53 is closely related to the poor prognosis of patients with pancreatic cancer, and the difference is of statistical significance. Although the study data does not support the ability of TP53 to regulate the extracellular secretion of pancreatic cancer by Rab27A, it is not possible to exclude the possibility of a weak correlation between the two. In the following study, the expression of Rab27B and TP53 in the specimens of 186 pancreatic cancer tissues, 51 corresponding adjacent tissues and 23 cases of benign lesions of the pancreas was detected by immunohistochemical method and tissue chip technique. The results showed that, The high expression of Rab27B can be used as an independent prognostic factor for pancreatic cancer. Through the Cox and multi-factor analysis, we found that the increase of the expression level of Rab27B is an important risk factor for the poor prognosis of the pancreatic cancer. The level of the expression of Rab27B and TP53 in the pancreatic cancer is also closely related to the vascular invasion and distant metastasis. The relationship between the expression of Rab27B and TP53 protein is also detected, and the relationship between TP53 and Rab27B remains to be further studied. In addition, the siRNA of Rab27A and Rab27B gene was constructed, and the cells of pancreatic cancer were transfected with the method of liposome, and the effects of the transfection and before and after transfection on the biological characteristics of the cells were detected by the methods of CCK-8, Transwell and Western Blot respectively. The results showed that the expression of Rab27A and Rab27B gene could inhibit the proliferation of pancreatic cancer, reduce its invasion ability and induce cell apoptosis. To sum up, this study first reported the high expression of Rab27A and TP53 in the tissues of pancreatic cancer and the high tissue of the pancreatic benign diseases, and the high expression of Rab27A was closely related to the poor prognosis of the patients with pancreatic cancer. The expression of Rab27B in pancreatic cancer has no significant difference with the expression of the adjacent tissue and the tissue of the pancreatic benign disease, but the five-year survival rate of Rab27B is significantly lower in patients with pancreatic cancer with high expression of Rab27B. We did not detect that Rab27A and TP53 are significantly related to the expression, but the expression of Rab27B and TP53 has a significant correlation, and the specific mode of action is to be further studied. The interference of Rab27A and Rab27B with siRNA technology can significantly reduce the proliferation, invasion, migration and apoptosis of pancreatic cancer cells, thus providing a theoretical basis and experimental basis for the next step to construct a novel targeted therapeutic agent.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R735.9
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1 王庆庆;Rab27A、Rab27B与TP53在胰腺癌中的作用及部分机制研究[D];苏州大学;2015年
，本文编号：2429539
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