兰州大学机构库 >学院待认领
体内及体外研究IL-6在肝细胞肝癌血管再生中的调控作用
Alternative TitleThe role of IL-6 in regulating HCC angiogenesis --In vivo and in vitro study
田爱平
Thesis Advisor严祥
2014-12-01
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name博士
Keyword肝细胞肝癌 白介素6 血管再生 索拉菲尼
Abstract目的:通过体内及体外研究探讨白介素6(IL-6)在原发性肝癌血管再生中的调节作用及机制,为原发性肝癌的抗血管再生靶向治疗提供新策略。 结果:C57小鼠DEN慢性诱导性肝癌组织中IL-6/STAT-3无论在分子水平还是蛋白水平,表达量均明显高于癌旁组织。同时肿瘤组织中血管再生相关因子(VEGF, VEGFR-2,CD31)蛋白水平均明显高于癌旁组织。结论:IL-6在小鼠及人HCC肿瘤组织中表达高于癌旁组织,且IL-6与VEGF,CD31及DLL4等血管再生因子表达水平高度正相关。体内及体外研究均表明调节IL-6表达水平后,肿瘤血管再生因子及细胞血管再生能力均相应被调节。上述结果证实了IL-6在肝癌患者肿瘤血管再生中的调控作用。本研究还发现IL-6/JAK-1/STAT-3信号通路可能介导索拉菲尼耐药机制。以上结果为我们进一步解决抗血管再生药物索拉菲尼耐药性问题及高表达IL-6 肝癌患者治疗提供了新的理论基础,也为肝癌患者的有效靶向化疗提供了新的思路和治疗策略。
Other AbstractAim: To investigate the role of IL-6 involves in regulating hepatocarcinoma angiogenesis and related mechanisms, and hopefully our study can provide new therapeutic strategy for anti-angiogenetic of HCC. In vivo study, we build mouse HCC xenograft animal model, and overexpressed IL-6 in the xenograft by transfecting plasmid IL-6, first of all, we need to confirm if IL-6 has been successfully overpressed through testing serum IL-6 by enzyme-linked immunosorbent assay (ELISA), after that, we used both RT-PCR and WB to to detect the gene and protein expression level of IL-6 and angiogenesis related factors and correlation rate between them in mouse HCC xenograft. In vitro study, we picked up several strong invation and metastatic HCC cell lines: HUH7, SNU182, SNU423 and PLC/PRF/5, and overexpressed IL-6 in these cells by thansfecting plasmid IL-6, after trasfection has been confirmed, we used pIL-6 conditioned medium (CM) of HCC cell lines to culture human umbilical vein endothelial cells (HUVEC), and observe HUVEC cell viability, cell proliferation, cell migration and angiogenetic function in pIL-6 CM through cell counting, bromodeoxyuridine (BrdU) assay, cell migration assay, and in vitro angiogenesis assay. After overexpression IL-6, we also inhibited IL-6 expression level in HCC cell lines by small interfering RNA (siRNA), and after the transfection has been confirmed, we detected HUVEC cell viability, cell proliferation, cell migration and angiogenetic function in siIL-6 CM through above assays. At last, we generated HUH7 sorafenib resistant cell lines, and test cell viability and IL-6/STAT3 signaling pathway expression level in sorafenib resistant cell lines and normal HUH7 cells. Results: both gene and protein expression level of IL-6/STAT3 was significantly higher in C57 DEN induced HCC tumor tissue compared with non-tumor. Meanwhile, angiogenetic factors (VEGF, VEGFR-2 and CD31) were also much higher in tumor tissue. According to ELISA and WB results, the correlation rate of IL-6 and VEGF protein expression level in human HCC tumor tissue was 0.928, from IHC results, we can tell that the correlation rate of IL-6 and vascular endothelium marker CD31 in human HCC tumor tissue was 0.951. After mouse HCC xenograft animal model was built up, and plasmid IL-6 was successfully transfected, the weight and the volume of the tumors were both significantly increased compared with control vector transfected tumor. Angiogenetic factors’ gene and protein e...
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222641
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
田爱平. 体内及体外研究IL-6在肝细胞肝癌血管再生中的调控作用[D]. 兰州. 兰州大学,2014.
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