IRF7在肝癌中的表达与其对肝癌细胞生物学行为的影响

发布时间：2018-02-21 21:09

本文关键词： 肝癌干扰素调节因子7 增殖周期侵袭　出处：《南昌大学》2016年硕士论文　论文类型：学位论文

【摘要】：研究目的:分析IRF7基因在肝癌与癌旁组织中的表达差异,并结合患者临床资料,探讨其表达与肝癌发生发展的关系;通过降低IRF7的表达,观察其对肝癌细胞生物学行为的影响,进一步了解IRF7基因影响肝癌进展的机制,从而为肝癌病变机制提供一定实验和理论依据。第一部分:IRF7在肝癌组织中的表达及与临床预后的关系实验目的:分析IRF7基因在肝癌及相应癌旁组织中的表达差异,并探讨其表达与肝癌临床预后的关系。实验方法:1、应用Western Blot及免疫组织化学方法检测IRF7在肝癌及相应癌旁组织中表达。2、结合患者临床资料,分析IRF7基因在肝癌组织中的表达与肝癌患者性别、年龄、血清AFP、肝硬化、肿瘤大小、肿瘤分期、肿瘤转移等临床相关特征之间的关系。实验结果:1、Western Blot检测结果:在所检测的40对肝癌组织及相应的癌旁肝组织中,29例(72.5%)肝癌组织中IRF7蛋白表达水平比相应的癌旁组织明显升高,11例(27.5%)癌旁组织中IRF7蛋白表达高于癌组织。2、免疫组织化学检测结果:在60例肝癌组织中IRF7高表达为65.0%(39/60),癌旁组织中高表达率为38.3%(23/60),IRF7在肝癌组织中高表达率明显高于癌旁组织高表达率,并具有统计学意义(P0.05)。3、分析IRF7的表达与各临床参数的关系,发现IRF7的表达与肿瘤的大小、转移呈正相关,与分化程度呈负相关。结论:IRF7基因在肝癌组织中高表达,且其表达与肝癌的侵袭和恶性程度相关,在肝癌的发生发展中起到重要的作用。第二部分:IRF7表达对肝癌细胞生物学功能的影响实验目的:通过细胞实验进一步研究IRF7基因的表达对肝癌细胞增殖、周期、侵袭的影响。实验方法:1、q RT-PCR和Western Blot方法检测正常肝脏细胞株HL-7702,及不同的肝癌细胞系(Hep G2,Huh-7,Hep3B、SMMC7721,MHCCLM3)中目的基因IRF7的表达。2、RNA干扰的方法特异性抑制了肝癌细胞中IRF7基因的表达,分别应用CCK8、流式细胞仪技术、Transwell法检测降低IRF7基因表达后对肝癌细胞增殖、细胞周期、侵袭转移能力的影响。实验结果:1、q RT-PCR与Western Blot结果提示,IRF7 mRNA和蛋白表达在各肝癌细胞系中的表达均比正常肝细胞高;且在肝癌细胞株MHCCLM3中IRF7表达较其他肝癌细胞系更高。2、RNA干扰MHCCLM3肝癌细胞株中IRF7的表达,qRT-PCR和Western Blot检测IRF7在mRNA和蛋白表达水平的干扰效果,并筛选出最佳的RNA干扰系列。3、CCK8检测、流式细胞仪技术、Transwell法检测发现,降低IRF7基因的表达,肝癌细胞增殖,周期及侵袭明显受到抑制。结论:IRF7基因在肝癌细胞株高表达。IRF7的表达能够促进肝癌细胞增殖、周期及侵袭。总结:本课题通过检测肝癌组织中IRF7的表达,并分析其与肝癌患者的临床资料,发现IRF7的表达与肝癌的发生发展密切相关。同时,通过细胞生物学功能实验,证实了IRF7的表达能够促进肝癌细胞增殖、周期、转移;进一步的揭示了IRF7基因的表达在肝癌的进展中发挥了重要的作用。
[Abstract]:Objective: to analyze the difference of IRF7 gene expression between HCC and paracancerous tissues, and to explore the relationship between the expression of IRF7 gene and the occurrence and development of HCC, and to observe the effect of IRF7 on the biological behavior of HCC cells. To further understand the mechanism of IRF7 gene affecting the progression of liver cancer, The first part is the expression of IRF7 gene in HCC tissues and its relationship with clinical prognosis. Objective: to analyze the difference of expression of IRF7 gene in HCC and its adjacent tissues. To investigate the relationship between the expression of IRF7 and the clinical prognosis of HCC, we used Western Blot and immunohistochemical method to detect the expression of IRF7 in HCC and its adjacent tissues, and combined with the clinical data of the patients. To analyze the expression of IRF7 gene in hepatocellular carcinoma (HCC) and its relationship with sex, age, serum AFP, liver cirrhosis, tumor size and tumor staging. The relationship between tumor metastasis and other clinicopathological characteristics. Results: the expression of IRF7 protein in 40 pairs of HCC tissues and corresponding adjacent liver tissues was higher than that in corresponding paracancerous tissues. The expression of IRF7 protein in adjacent tissues was significantly higher than that in cancer tissues. The results of immunohistochemistry: in 60 cases of liver cancer, the high expression rate of IRF7 was 65.039 / 60%, and the high expression rate of 38.3% / 2360% IRF7 was higher in HCC tissues. The rate of high expression was significantly higher than that of paracancerous tissues. It was found that the expression of IRF7 was positively correlated with tumor size and metastasis, and negatively correlated with the degree of differentiation. Conclusion the expression of IRF7 gene is highly expressed in hepatocellular carcinoma. And its expression is related to the invasion and malignancy of liver cancer. The second part is the effect of the expression of IRF7 7 on the biological function of hepatoma cells. Objective: to further study the effect of IRF7 gene expression on the proliferation and cycle of hepatoma cells through cell experiments. The expression of IRF7 gene in HL-7702 cells and different hepatoma cell lines Hep G2H Huh-7h-7, Hep3BSS-SMMC7721 MHCCLM3 was detected by using the methods of 1: 1Q RT-PCR and Western Blot, which specifically inhibited the expression of IRF7 gene in hepatoma cells. CCK8 and flow cytometry were used to detect the proliferation and cell cycle of hepatoma cells after IRF7 gene expression was reduced. The results showed that the expression of IRF7 mRNA and protein in HCC cells was higher than that in normal hepatocytes. Moreover, the expression of IRF7 in HCC cell line MHCCLM3 was higher than that in other HCC cell lines. The expression of IRF7 in MHCCLM3 cell line was interfered with qRT-PCR and Western Blot were used to detect the interference effect of IRF7 on mRNA and protein expression, and the best RNA interference series. Flow cytometry analysis showed that the expression of IRF7 gene decreased, the proliferation, cycle and invasion of HCC cells were inhibited obviously. Conclusion the high expression of% IRF7 gene in hepatoma cells can promote the proliferation of HCC cells. Period and invasion. Summary: by detecting the expression of IRF7 in HCC and analyzing the clinical data of HCC patients, we found that the expression of IRF7 is closely related to the occurrence and development of HCC. At the same time, through the experiment of cell biological function, we found that the expression of IRF7 is closely related to the occurrence and development of HCC. It is confirmed that the expression of IRF7 can promote the proliferation, cycle and metastasis of HCC cells, and further reveal that the expression of IRF7 gene plays an important role in the progression of HCC.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.7

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