【作者】 贾慧丽；

【导师】 徐芸；

【作者基本信息】 郑州大学 ， 内科学， 2013， 硕士

【摘要】 研究背景肝细胞肝癌(hepatocellular carcinoma, HCC)是我国最常见的肿瘤之一。目前,虽然对肝癌的癌变机制取得一定的进展,然而具体的分子生物学机制尚不清楚。肝癌的发生发展涉及多基因、多因素、多步骤的连续过程。Argonaute(AGO)蛋白家族的成员之一PIWIL2,在多种癌组织中表达,然而,PIWIL2在HCC中的研究尚未报道。本研究通过RT-PCR、Western-blot方法分别检测PIWIL2mRNA及蛋白在肝癌及其对应的癌旁组织中的表达并结合临床资料进行分析,寻求肿瘤治疗的靶点,为肿瘤筛选及预后提供生物学依据。研究目的检测肝癌组织中PIWIL2基因mRNA和蛋白的表达,进而探讨PIWIL2基因在临床诊断中的意义。研究方法1.70例肝细胞肝癌及其癌灶旁(>2am)的非癌组织,均经病理学证实。分别来自2011年1月-2012年3月郑州大学第一附属医院手术切除标本,所有患者术前均未实施过其它治疗。2.采用反转录一聚合酶链反应(RT-PCR)和Western-blot印迹技术,检测了70例肝癌及其对应癌旁组织中PiWi12基因mRNA及其蛋白的表达并结合临床资料进行分析。3.采用SPSS13.0统计软件进行统计学处理。数据处理两样本率的比较采用四格表资料的Z检验,两样本定量资料采用检验,以P<0.05为差异显著性的标准。研究结果PiWi12mRNA在肝癌中的表达量相对值(0.91±0.04)高于癌旁的肝脏组织的表达量(0.32±0.04)(P<0.05)。Piwi12基因mRNA的表达与患者的肝内转移、分化程度及癌栓等病理因素方面的差异有统计学意义(P＜0.05)。蛋白在肝癌中的表达均高于对应的癌旁组织。Piwi12的蛋白质表达与患者的肝内转移及癌栓等病理因素方面的差异有统计学意义(P<0.05)。 Piwi12基因mRNA的表达与Piwi12的蛋白质的表达有相关性(P＜0.05)。结论1.Piwi12表达与肝内转移及门静脉癌栓有关,可能为HCC候选癌基因之一。2.Piwi12有可能成为肝癌的检测及治疗的一种分子标志物。更多还原

【Abstract】 BackgroundHepatocellular carcinoma(HCC) is a common tumor in China. At present, although the mechanism of carcinogenesis of hepatocellular carcinoma makes some progress, the specific molecular mechanisms remains unclear. HCC is a long-term complex process in which multi-stage, multi-factor and multiple genes are involved. PIWI(P-element induced wimpy testis) domain (PPD) proteins are members of a well-conserved family which is expressed in a variety of organisms. However, there is not any evidence to show that the relationship between status of mRNA of PIWIL2and expression of its protein in HCC. We use RT-PCR and Western-blot to discuss their role as a cancer gene in the invasion and metastasis of the HCC which will provide new insights into HCC carcinogenesis, cancer treatment and feasible chemopreventive measures for the future.ObjectTo investigate the mRNA and protein expression of Piwil2gene in hepatocellular carcinoma(HCC), and then discuss the clinical significance of Piwil2in hepatocellular carcinoma(HCC).Methods1.70cases of HCC specimens and matched tumor-adjacent non-cancerous specimens had been collected from January2011to April2012in the first affiliated hospital of Zhengzhou University. All cases are diagnosed by experienced pathological doctor and are not accepted any treatment before operation.2. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the Piwil2mRNA and protein levels in70Surgical excised HCC specimens and paired adjacent non-cancerous specimens.3SPSS13.0statistical software is used for statistical analysis. The relative quantitative RT-PCR results were expressed as mean and standard deviation and tested for statistical significance with Student’s two-tailed t-test. Significant differences between the expression of Runx3and clinicopathological parameters were compared by the X2-test.All P-values were derived from two-sided tests and a probability less than0.05was considered statistically significant.ResultsThe relative expression value of Piwil2mRNA was0.91±0.04in HCC vs0.32±0.04in adjacent non-cancerous tissues (t=37.240, P<0.05). The Piwil2mRNA expression in HCC correlated with some clinical pathological parameters including low tumor differentiation, intrahepatic invasion and metastasis. The protein expression of Piwil2gene in HCC was higher than that in adjacent non-cancerous tissues. The Piwil2protein expression in HCC correlated with some clinical pathological parameters including intrahepatic invasion and metastasis. Furthermore there was a distinct correlation between the mRNA and protein expression of Piwi2gene(x2=13.113P<0.01).Conclusions1. Piwi2expression was releated to the intrahepatic invasion and metastasis and may be one of candidate cancer genes.2. The Piwil2had the potential to be developed into a novel biomarker for detection and therapy of HCC.更多还原