【摘要】：目的:構(gòu)建RNAi干擾肝癌細(xì)胞株HePG-2,通過(guò)實(shí)驗(yàn)觀察各種肝素酶不同表達(dá)水平的肝癌細(xì)胞對(duì)血管內(nèi)皮細(xì)胞粘附分子表達(dá)的影響,完善并豐富肝素酶在肝癌侵襲轉(zhuǎn)移中的分子學(xué)研究。方法:(1)選用肝癌細(xì)胞株HePG-2作為研究的實(shí)驗(yàn)細(xì)胞,運(yùn)用抑制肝素酶的基因?qū)ePG-2進(jìn)行細(xì)胞轉(zhuǎn)染,通過(guò)qRT-PCR及Western-blot等實(shí)驗(yàn)方法檢測(cè)各組轉(zhuǎn)染細(xì)胞中HPSE基因表達(dá)的水平;(2)通過(guò)上述方法選擇出相應(yīng)的HePG-2細(xì)胞分為轉(zhuǎn)染組(RNAi干擾的HePG-2細(xì)胞)、陰性對(duì)照組(轉(zhuǎn)染陰性的HePG-2細(xì)胞)、空白對(duì)照組(未轉(zhuǎn)染的HePG-2細(xì)胞)三組實(shí)驗(yàn)細(xì)胞進(jìn)行細(xì)胞粘附實(shí)驗(yàn),通過(guò)qRT-PCR及Western-blot等實(shí)驗(yàn)方法分析各實(shí)驗(yàn)組細(xì)胞粘附分子表達(dá)的影響。結(jié)果:(1)運(yùn)用qRT-PCR及Western-blot等實(shí)驗(yàn)方法分析刷選,成功得到最佳干擾的HePG-2細(xì)胞;(2)qRT-PCR和Western-blot結(jié)果:未轉(zhuǎn)染的HePG-2細(xì)胞對(duì)血管內(nèi)皮細(xì)胞粘附分子表達(dá)有明顯增強(qiáng);RNAi干擾的HePG-2細(xì)胞對(duì)血管內(nèi)皮細(xì)胞粘附分子表達(dá)有明顯降低;轉(zhuǎn)染陰性的HePG-2細(xì)胞對(duì)血管內(nèi)皮細(xì)胞粘附分子表達(dá)相對(duì)空白對(duì)照組有所降低,但高于RNAi干擾的HePG-2細(xì)胞組。結(jié)論:通過(guò)qRT-PCR及Western-blot等實(shí)驗(yàn)方法證實(shí)肝素酶對(duì)肝癌細(xì)胞在與血管內(nèi)皮細(xì)胞粘附的作用過(guò)程中發(fā)揮了相關(guān)重要的的作用,參與介導(dǎo)了肝癌細(xì)胞與血管內(nèi)皮細(xì)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.
【學(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

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 趙云峰,王英年,劉加洪,姚艷紅,張可佳;P-選擇素與腫瘤轉(zhuǎn)移[J];中華結(jié)核和呼吸雜志;2001年09期

3 劉芳,孔旭黎,樊紅亮,高冰;P-選擇素與腫瘤轉(zhuǎn)移關(guān)系的研究[J];現(xiàn)代醫(yī)藥衛(wèi)生;2005年20期

4 陳孝平,裘法祖,吳在德,何松青,張萬(wàn)廣,劉安重,孟春城,江斌,喬森,潘華鋒,史光軍,曹斌;肝細(xì)胞癌門(mén)靜脈癌栓形成的分子生物學(xué)機(jī)制研究[J];中華實(shí)驗(yàn)外科雜志;2005年09期

5 李燕杰;李淑蓮;;乙酰肝素酶的分子特性及其在生理病理過(guò)程中的作用[J];河南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年03期

6 范磊;吳強(qiáng);邢曉娟;劉郁東;邵增務(wù);;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 陳曉鵬;羅俊生;田野;聶宸麟;崔巍;張衛(wèi)東;;肝素酶調(diào)節(jié)對(duì)肝癌細(xì)胞粘附侵襲能力的影響[J];中國(guó)腫瘤臨床;2014年18期

8 王煉煉;鄧幼林;李聰;;Chk1基因?qū)θ寺殉舶┘?xì)胞HO8910增殖及細(xì)胞周期調(diào)控的影響及可能機(jī)制[J];重慶醫(yī)科大學(xué)學(xué)報(bào);2014年11期

9 干惠珠;張學(xué)文;孫步彤;王世寶;;乙酰肝素酶蛋白表達(dá)與肝癌侵襲轉(zhuǎn)移關(guān)系的研究[J];中國(guó)實(shí)驗(yàn)診斷學(xué);2014年09期

10 師騰瑞;于衛(wèi)華;海春旭;秦緒軍;;活性氧調(diào)控細(xì)胞增殖的分子機(jī)制研究進(jìn)展[J];癌變·畸變·突變;2015年06期

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