KLF17在肝癌中的作用及对肝癌预后的影响

发布时间：2018-01-25 18:25

本文关键词： 肝癌KLF17迁移侵袭预后　出处：《北京协和医学院》2016年博士论文　论文类型：学位论文

【摘要】：目的：肝癌已成为全球第五种最常见的恶性肿瘤,在中国肝癌发病率水平男性居第5位,女性居第6位；死亡率水平男性居第2位,女性居第5位。我国肝癌患者多数有肝炎、肝硬化病史,临床80%左右患者因各种原因不能手术,而且术后高复发,高转移,成为肝癌患者治疗中难以解决的问题。而恶性肿瘤侵袭和转移的能力与上皮-间质转化(Epithelial-Mesenchymal Transition, EMT)密切相关,我们计划研究肝癌细胞的EMT调控机制,为肝癌治疗提供新的可能的治疗靶点。Hepll和Hep12是从同一肝癌患者提取的两株分别具有上皮或者间质样细胞性质的肿瘤细胞,Hepl2的迁移和侵袭能力显著高于Hep11,而在Hep11中转录因子KLF17表达显著高于Hepl2,在乳腺癌,胃癌,食道癌中KLF17都是抑癌基因,所以我们计划研究KLF17在肝癌细胞中作用及作用机制,以及KLF17的表达和肝细胞癌患者预后的关系。方法：利用SiRNA技术干扰KLF17在肝癌细胞系中的表达,然后利用Transwell法检测KLF17干扰对肝癌细胞系迁移和侵袭能力的影响。利用Western Blot口定量PCR技术检测KLF17干扰后上皮和间质相关基因在mRNA和蛋白水平的表达,明确KLF17对上皮间质转化和迁移侵袭的影响。选择2009年1月至2011年12月在北京协和医院肝胆外科手术的肝细胞肝癌患者,有完整随访资料的患者,免疫组化法检测KLF17在肝细胞癌组织中的表达水平,评估其和患者DFS,OS之间的关系。结果：研究发现在3个肝癌细胞系中,KLF17干扰后细胞系的迁移和侵袭能力显著增加,同时发现其上皮基因表达下调,间质基因表达上调。在36名可评估的肝癌患者中其中有19人(52.8%)为KLF17高表达,17人(47.2%)低表达。Cox回归分析发现巴塞罗那分期和KLF17表达水平对DFS和OS有影响。单因素分析KLF17对预后的影响,发现KLF17免疫组化低表达的患者的DFS为23.4±4.9个月(95%可信区间为13.2-32.3个月),高表达的DFS为39.3±4.2个月(95%可信区间为31.1~47.5个月),p0.05。KLF17评分为低表达患者的OS为35.2±4.2个月(95%可信区间27.0-43.4个月)高表达患者的OS为46.9±3.2(95%可信区间40.1~53.2个月),KLF17高表达和低表达的肝细胞肝癌患者DFS和OS均有显著性差异(p0.05)。结论：本研究发现KLF17调控肝癌细胞迁移和侵袭,可能是通过影响上皮间质转化；在肝细胞癌患者中,KLF17表达高是独立的预后较好的因素。
[Abstract]:Objective: hepatocellular carcinoma (HCC) has become the most common malignant tumor in the world. The incidence of HCC in China is the fifth in males and the 6th in females. The mortality level was the second in males and the 5th in females. The majority of HCC patients in China had hepatitis and cirrhosis history. About 80% patients could not be operated on for various reasons and had high recurrence and high metastasis after operation. The ability of malignant tumor invasion and metastasis and epithelial-interstitial transformation (EMT) are difficult to solve in patients with liver cancer. Epithelial-Mesenchymal Transition. We plan to study the mechanism of EMT regulation in hepatoma cells. Hepll and Hep12 are two tumor cells with epithelial or interstitial cell nature extracted from the same HCC patients. The migration and invasion ability of Hepl2 was significantly higher than that of Hep11, while the expression of transcription factor KLF17 in Hep11 was significantly higher than that in Hepl2, and in breast cancer and gastric cancer. KLF17 is a tumor suppressor gene in esophageal cancer, so we plan to study the role and mechanism of KLF17 in liver cancer cells. Methods: SiRNA technique was used to interfere the expression of KLF17 in hepatocellular carcinoma cell line. Then the effect of KLF17 interference on the migration and invasion of hepatoma cell lines was detected by Transwell assay. Blot quantitative PCR was used to detect the expression of epithelial and interstitial genes in mRNA and protein after KLF17 interference. To determine the effect of KLF17 on epithelial interstitial transformation and migration and invasion, we selected patients with hepatocellular carcinoma (HCC) who underwent hepatobiliary surgery from January 2009 to December 2011 in Peking Union Hospital. Immunohistochemical method was used to detect the expression of KLF17 in hepatocellular carcinoma (HCC) and to evaluate the expression of KLF17 and DFS in the patients with complete follow-up data. Results: it was found that the migration and invasion of three hepatoma cell lines were increased after KLF17 interference, and the expression of epithelial genes was down-regulated. The expression of mesenchymal genes was up-regulated. Among the 36 patients with liver cancer, 19 (52.8%) had high expression of KLF17. Low expression. Cox regression analysis showed that Barcelona stage and KLF17 expression level had influence on DFS and OS. Univariate analysis showed that KLF17 had influence on prognosis. It was found that the DFS of the patients with low expression of KLF17 was 23.4 卤4.9 months and 95% confidence interval was 13.2-32.3 months. The DFS with high expression was 39.3 卤4.2 months and 95% confidence interval was 31.1-47.5 months). The OS of low expression patients with p0.05.KLF17 score was 35.2 卤4.2 months and 95% confidence interval was 27.0-43.4 months. The OS of high expression patients was 46.9 卤3.2 95% confidence interval (40.1 ~ 53.2 months). There was significant difference between DFS and OS in HCC patients with high and low expression of KLF17. Conclusion: in this study, KLF17 regulates the migration and invasion of HCC cells. It may be through the influence of epithelial mesenchymal transformation; High expression of KLF17 is an independent prognostic factor in hepatocellular carcinoma.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7

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