肝素酶对血管内皮细胞粘附分子P选择素表达的影响

发布时间：2018-11-09 14:32

【摘要】：目的:构建RNAi干扰肝癌细胞株HePG-2,通过实验观察各种肝素酶不同表达水平的肝癌细胞对血管内皮细胞粘附分子表达的影响,完善并丰富肝素酶在肝癌侵袭转移中的分子学研究。方法:(1)选用肝癌细胞株HePG-2作为研究的实验细胞,运用抑制肝素酶的基因对HePG-2进行细胞转染,通过qRT-PCR及Western-blot等实验方法检测各组转染细胞中HPSE基因表达的水平;(2)通过上述方法选择出相应的HePG-2细胞分为转染组(RNAi干扰的HePG-2细胞)、阴性对照组(转染阴性的HePG-2细胞)、空白对照组(未转染的HePG-2细胞)三组实验细胞进行细胞粘附实验,通过qRT-PCR及Western-blot等实验方法分析各实验组细胞粘附分子表达的影响。结果:(1)运用qRT-PCR及Western-blot等实验方法分析刷选,成功得到最佳干扰的HePG-2细胞;(2)qRT-PCR和Western-blot结果:未转染的HePG-2细胞对血管内皮细胞粘附分子表达有明显增强;RNAi干扰的HePG-2细胞对血管内皮细胞粘附分子表达有明显降低;转染阴性的HePG-2细胞对血管内皮细胞粘附分子表达相对空白对照组有所降低,但高于RNAi干扰的HePG-2细胞组。结论:通过qRT-PCR及Western-blot等实验方法证实肝素酶对肝癌细胞在与血管内皮细胞粘附的作用过程中发挥了相关重要的的作用,参与介导了肝癌细胞与血管内皮细胞的粘附。
[Abstract]:Objective: to construct a RNAi interfering hepatoma cell line HePG-2, and observe the effect of different levels of heparinase expression on the expression of adhesion molecules in vascular endothelial cells (VECs). To improve and enrich the molecular studies of heparanase in the invasion and metastasis of liver cancer. Methods: (1) Hepatocellular carcinoma cell line HePG-2 was selected as the experimental cell, and HePG-2 was transfected with the gene that inhibited heparinase. QRT-PCR and Western-blot were used to detect the expression of HPSE gene in the transfected cells. (2) the corresponding HePG-2 cells were selected to be divided into transfection group (RNAi interfering HePG-2 cells) and negative control group (transfected negative HePG-2 cells). Three groups of untransfected HePG-2 cells in blank control group were tested for cell adhesion. The effects of cell adhesion molecules on the expression of adhesion molecules were analyzed by means of qRT-PCR and Western-blot. Results: (1) the best interfering HePG-2 cells were successfully obtained by qRT-PCR and Western-blot. (2) the results of qRT-PCR and Western-blot: the expression of vascular endothelial cell adhesion molecules was significantly increased in untransfected HePG-2 cells, and decreased by RNAi interfering HePG-2 cells. Compared with the blank control group, the expression of adhesion molecules in the transfected HePG-2 cells was lower than that in the control group, but higher than that in the HePG-2 cells with RNAi interference. Conclusion: heparanase plays an important role in the adhesion of HCC cells to vascular endothelial cells by means of qRT-PCR and Western-blot. It plays an important role in mediating the adhesion of hepatoma cells to vascular endothelial cells.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

【参考文献】

相关期刊论文 前10条

1 ;Interaction of vascular endothelial cells with leukocytes, platelets and cancer cells in inflammation, thrombosis and cancer growth and metastasis[J];Acta Pharmacologica Sinica;2003年12期

2 赵云峰,王英年,刘加洪,姚艳红,张可佳;P-选择素与肿瘤转移[J];中华结核和呼吸杂志;2001年09期

3 刘芳,孔旭黎,樊红亮,高冰;P-选择素与肿瘤转移关系的研究[J];现代医药卫生;2005年20期

4 陈孝平,裘法祖,吴在德,何松青,张万广,刘安重,孟春城,江斌,乔森,潘华锋,史光军,曹斌;肝细胞癌门静脉癌栓形成的分子生物学机制研究[J];中华实验外科杂志;2005年09期

5 李燕杰;李淑莲;;乙酰肝素酶的分子特性及其在生理病理过程中的作用[J];河南大学学报(医学版);2008年03期

6 范磊;吴强;邢晓娟;刘郁东;邵增务;;Targeted Silencing of Heparanase Gene by Small Interfering RNA Inhibits Invasiveness and Metastasis of Osteosarcoma Cells[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2011年03期

7 陈晓鹏;罗俊生;田野;聂宸麟;崔巍;张卫东;;肝素酶调节对肝癌细胞粘附侵袭能力的影响[J];中国肿瘤临床;2014年18期

8 王炼炼;邓幼林;李聪;;Chk1基因对人卵巢癌细胞HO8910增殖及细胞周期调控的影响及可能机制[J];重庆医科大学学报;2014年11期

9 干惠珠;张学文;孙步彤;王世宝;;乙酰肝素酶蛋白表达与肝癌侵袭转移关系的研究[J];中国实验诊断学;2014年09期

10 师腾瑞;于卫华;海春旭;秦绪军;;活性氧调控细胞增殖的分子机制研究进展[J];癌变·畸变·突变;2015年06期

，

本文编号：2320687