兰州大学机构库 >学院待认领
AFP对人肝癌细胞生物学特性及对VEGF 信号通路的影响
Alternative TitleThe effect of AFP to the cellular property and VEGF signal pathway in human HCC
孟文勃
Thesis Advisor李玉民
2011-12-01
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword肝细胞癌 Huh-7细胞系 甲胎蛋白 VEGF 血管生成 侵袭
Abstract肝细胞癌是世界上最常见的恶性肿瘤之一,全世界每年50-60万例新发肝癌中约50%发生在我国。肝癌的发生发展在分子水平是多通路、网络性交错作用的过程,因此,多靶点、多抑制剂药物的开发已成为肝癌药物研发的趋势。甲胎蛋白是分子量为68,000的单肽,同时也是迄今为止在HCC中发现的最重要的肿瘤标记物。临床发现绝大多数HCC患者合并有高AFP表达。AFP在细胞增殖及凋亡的多个路径中起调节作用。VEGF / VEGFR信号是胚胎发育和病理模型中血管生成、再生和淋巴管生成过程的中心环节。VEGFR-2介导VEGF信号通过诱导内皮细胞增殖、迁移以及增加内皮通透性刺激血管生成。本研究主要对人肝癌Huh-7细胞株和HepG2细胞株进行体外培养,利用siRNA干扰技术下调肝癌细胞中AFP的表达。结果显示,Huh-7细胞株呈AFP阳性、HepG2细胞株呈AFP阴性。因此,本实验利用Huh-7细胞株为研究对象,进行后续实验。流式结果显示,与对照组相比,行siRNA/ AFP干扰后的Huh-7细胞株,细胞生长周期多停留在G1期。TUNEL细胞凋亡检测显示,转染组中出现了大量的荧光凋亡信号。Western blot显示,Huh-7细胞株行AFP siRNA转染后VEGF及VEGFR2低表达,而对照组和LP2000组高表达。利用激光共聚焦显微镜进一步研究证实了Western Blot的研究结果。AFP siRNA转染后MMP2和MMP9均低表达。通过以上实验结果,本研究发现,肝癌肿瘤标记物AFP直接参与VEGF/VEGFR信号通路,AFP表达水平与VEGF/VEGFR表达呈正相关,AFP通过调控VEGF/VEGFR信号通路调控肝癌生物学行为。
Other AbstractHepatocellular carcinoma is the third most common cause of cancer- related death worldwide with approximately 600,000 patients dying from this disease annually.The tumorigenesis is a multi and complex pathway process. So, the drug creation against multi-molecular targets and inhibitors is a new focus in clinical research. Alpha-fetoprotein (AFP) is frequently found in HCC (positive in more than 70% of tumors) and is considered to be a diagnostic and prognostic tumor marker. AFP is well known as a tumor maker in ontogenesis and tumorigenesis. It plays an important role in many pathways of cellular apoptosis and hepatocellular carcinoma progress. VEGF/VEGFR signaling pathway has been identified as the center of vasculogenesis, revasculorization and lymphatic generation in embryogenesis and pathological models. The VEGF signal mediated by VEGFR2 stimulates vasculogenesis by inducing proliferation, generation and migration of endothelinum, and improving the transparency of endoepithelium.In our research, the Huh-7 and HepG2 cell lines were cultured in vitro. The AFP expression was down regulated by using siRNA and gene silence.Compared with control, the cell cycle of most cells transfected with AFP siRNA stayed in G1 stage. The apoptosis data shows lots of apoptotic cell marked with fluorescence appeared.The western blot data show VEGF, VEGFR2, MMP2 and MMP9 down expression in AFP siRNA transfected Huh-7 cell line compared with control. The results were confirmed by confocal microscopy. It shows that the AFP may play an important role in VEGF/VEGFR2 signaling pathway, as well in tumor invasion.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/224245
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
孟文勃. AFP对人肝癌细胞生物学特性及对VEGF 信号通路的影响[D]. 兰州. 兰州大学,2011.
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