本文選題：鼻咽腫瘤 + 蛋白酶活化受體-1　； 參考：《華中科技大學(xué)》2010年碩士論文

【摘要】：目的:檢測(cè)蛋白酶活化受體1 (PAR1)在鼻咽癌細(xì)胞中的表達(dá),研究其在鼻咽癌細(xì)胞增殖、黏附和侵襲等惡性生物學(xué)行為中的作用。 方法:選取5株有代表性的鼻咽癌細(xì)胞系,其中CNE1為高分化細(xì)胞系,CNE1-LMP1為穩(wěn)定轉(zhuǎn)染潛伏膜蛋1 (LMP1)基因的細(xì)胞系,其已被證實(shí)比CNE1細(xì)胞系更具侵襲性和轉(zhuǎn)移性,CNE2為低分化細(xì)胞系,SUNE1-5-8F為高成瘤高轉(zhuǎn)移細(xì)胞系,而SUNE1-6-10B為只成瘤不轉(zhuǎn)移細(xì)胞系,Real time PCR和Western blot檢測(cè)5株鼻咽癌細(xì)胞系中PAR1 mRNA和蛋白的表達(dá);細(xì)胞免疫熒光法檢測(cè)PAR1在鼻咽癌細(xì)胞中的亞細(xì)胞定位;MTT、黏附實(shí)驗(yàn)和Transwell侵襲實(shí)驗(yàn)分別用來(lái)檢測(cè)PAR1特異性激動(dòng)肽SFLLRN和凝血酶對(duì)CNE1-LMP1細(xì)胞增殖、黏附和體外侵襲的影響;明膠酶譜法檢測(cè)SFLLRN和凝血酶對(duì)CNE1-LMP1細(xì)胞培養(yǎng)上清液中MMP2和MMP9活性的影響。 結(jié)果:PAR1在5株鼻咽癌細(xì)胞系中都存在表達(dá),其主要分布在細(xì)胞膜和細(xì)胞漿, CNE1-LMP1和SUNE1-5-8F中PAR1的表達(dá)量明顯多于CNE1、CNE2和SUNE1-6-10B,而CNE1-LMP1細(xì)胞中PAR1的表達(dá)量為5株細(xì)胞中最多一株。SFLLRN能夠促進(jìn)CNE1-LMP1細(xì)胞增殖、黏附和體外侵襲。凝血酶亦能提高CNE1-LMP1細(xì)胞的黏附和體外侵襲能力,但其對(duì)細(xì)胞的增殖起雙重作用,0.1~0.5U/ml的凝血酶能夠促進(jìn)細(xì)胞的增殖,而高于0.5U/ml的凝血酶則抑制細(xì)胞的增殖。CNE1-LMP1細(xì)胞同凝血酶共孵育24小時(shí),其細(xì)胞培養(yǎng)上清液中MMP2和MMP9的活性明顯升高;與凝血酶相比,SFLLRN僅能夠輕微地上調(diào)CNE1-LMP1細(xì)胞中MMP2的活性,而對(duì)MMP9的活性影響甚微。 結(jié)論:PAR1在鼻咽癌細(xì)胞中的表達(dá)與鼻咽癌細(xì)胞的侵襲轉(zhuǎn)移潛能相關(guān);PAR1的活化能夠提高鼻咽癌細(xì)胞的增殖、黏附和侵襲能力,并在一定程度上促進(jìn)細(xì)胞明膠酶的分泌。
[Abstract]:Aim: to investigate the expression of protease activated receptor 1 (PAR1) in nasopharyngeal carcinoma (NPC) cells and its role in malignant biological behaviors such as proliferation, adhesion and invasion of nasopharyngeal carcinoma cells.It has been proved that CNE _ 2 is more invasive and metastatic than CNE1 cell line. SUNE1-5-8F, a poorly differentiated cell line, is a highly metastatic cell line of Gao Cheng tumor, while SUNE1-6-10B is a non-metastasis cell line. Real time PCR and Western blot are used to detect the expression of PAR1 mRNA and protein in five nasopharyngeal carcinoma cell lines.The subcellular localization of PAR1 in nasopharyngeal carcinoma cells was detected by cell immunofluorescence assay. The effects of PAR1 specific activator SFLLRN and thrombin on the proliferation, adhesion and invasion of CNE1-LMP1 cells in vitro were detected by adhesion assay and Transwell invasion assay, respectively.The effects of SFLLRN and thrombin on the activities of MMP2 and MMP9 in the supernatant of CNE1-LMP1 cell culture were detected by gelatinase assay.Results all of the 5 nasopharyngeal carcinoma cell lines were expressed in the presence of 1% PAR1.The expression of PAR1 in CNE1-LMP1 and SUNE1-5-8F was significantly higher than that in CNE1CNE2 and SUNE1-6-10B, while the expression of PAR1 in CNE1-LMP1 cells was more than that in 5 cell lines. SFLLRN could promote the proliferation, adhesion and invasion of CNE1-LMP1 cells in vitro.Thrombin could also enhance the adhesion and invasiveness of CNE1-LMP1 cells in vitro, but it had double effects on cell proliferation.However, there was little effect on the activity of MMP9.Conclusion the expression of 1% PAR1 in nasopharyngeal carcinoma cells is related to the invasion and metastasis potential of nasopharyngeal carcinoma cells. The activation of PAR1 can enhance the proliferation, adhesion and invasion ability of nasopharyngeal carcinoma cells, and to some extent promote the secretion of gelatinase in nasopharyngeal carcinoma cells.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.63

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 張立華;蔣星軍;馮湘玲;劉衛(wèi)東;張樹(shù)菊;任彩萍;;蛋白酶活化受體1在不同腫瘤中的研究進(jìn)展[J];生命科學(xué)研究;2012年03期

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1 張立華;Flotillin-2在鼻咽癌中的表達(dá)及作用機(jī)制的初步探索[D];中南大學(xué);2012年

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本文編號(hào)：1740668