蛋白酶活化受体1在鼻咽癌细胞中的表达及其意义

发布时间：2018-04-12 17:13

本文选题：鼻咽肿瘤 + 蛋白酶活化受体-1　；参考：《华中科技大学》2010年硕士论文

【摘要】：目的:检测蛋白酶活化受体1 (PAR1)在鼻咽癌细胞中的表达,研究其在鼻咽癌细胞增殖、黏附和侵袭等恶性生物学行为中的作用。方法:选取5株有代表性的鼻咽癌细胞系,其中CNE1为高分化细胞系,CNE1-LMP1为稳定转染潜伏膜蛋1 (LMP1)基因的细胞系,其已被证实比CNE1细胞系更具侵袭性和转移性,CNE2为低分化细胞系,SUNE1-5-8F为高成瘤高转移细胞系,而SUNE1-6-10B为只成瘤不转移细胞系,Real time PCR和Western blot检测5株鼻咽癌细胞系中PAR1 mRNA和蛋白的表达;细胞免疫荧光法检测PAR1在鼻咽癌细胞中的亚细胞定位;MTT、黏附实验和Transwell侵袭实验分别用来检测PAR1特异性激动肽SFLLRN和凝血酶对CNE1-LMP1细胞增殖、黏附和体外侵袭的影响;明胶酶谱法检测SFLLRN和凝血酶对CNE1-LMP1细胞培养上清液中MMP2和MMP9活性的影响。结果:PAR1在5株鼻咽癌细胞系中都存在表达,其主要分布在细胞膜和细胞浆, CNE1-LMP1和SUNE1-5-8F中PAR1的表达量明显多于CNE1、CNE2和SUNE1-6-10B,而CNE1-LMP1细胞中PAR1的表达量为5株细胞中最多一株。SFLLRN能够促进CNE1-LMP1细胞增殖、黏附和体外侵袭。凝血酶亦能提高CNE1-LMP1细胞的黏附和体外侵袭能力,但其对细胞的增殖起双重作用,0.1~0.5U/ml的凝血酶能够促进细胞的增殖,而高于0.5U/ml的凝血酶则抑制细胞的增殖。CNE1-LMP1细胞同凝血酶共孵育24小时,其细胞培养上清液中MMP2和MMP9的活性明显升高;与凝血酶相比,SFLLRN仅能够轻微地上调CNE1-LMP1细胞中MMP2的活性,而对MMP9的活性影响甚微。结论:PAR1在鼻咽癌细胞中的表达与鼻咽癌细胞的侵袭转移潜能相关;PAR1的活化能够提高鼻咽癌细胞的增殖、黏附和侵袭能力,并在一定程度上促进细胞明胶酶的分泌。
[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.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63

【引证文献】