COX-2、MMP-14和胰腺癌细胞增殖、侵袭、转移的关系研究

发布时间：2018-01-13 14:11

本文关键词：COX-2、MMP-14和胰腺癌细胞增殖、侵袭、转移的关系研究　出处：《石河子大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的:通过测定不同浓度的环氧合酶-2抑制剂塞来昔布对胰腺癌细胞增殖、侵袭、迁移能力以及对环氧合酶-2(cyclooxygenase-2,COX-2)、基质金属蛋白酶14(matrix metalloproteinase14,MMP-14)的蛋白表达影响来探讨COX-2抑制剂塞来昔布对胰腺癌的作用及相关作用机制。方法:1.体外培养人胰腺癌细胞系PANC-1细胞;2.分别用0(无)、20(低)、60(中)、100(高)μmol/L四个塞来昔布浓度对PANC-1细胞进行不同时间(24、48和72h)处理,MTT比色法检测细胞的增殖能力,Transwell侵袭实验和划痕实验检测细胞的侵袭能力和迁移能力,酶联免疫吸附剂测定(EILISA)法检测COX-2和MMP-14的蛋白表达情况;3.数据进行统计分析。结果:1.MTT结果显示不同浓度的塞来昔布作用24、48和72h后PANC-1细胞的增殖能力下降,随着塞来昔布浓度的增加,抑制作用逐渐增强,且随着塞来昔布作用时间的延长,抑制作用也逐渐加强,抑制作用呈时间和浓度依赖性(P0.05);2.Transwell侵袭实验结果显示不同浓度的塞来昔布作用24h后PANC-1细胞的侵袭能力下降,穿过基质胶的细胞个数分别为300.654±12.558、271.041±10.569、184.003±10.865和92.667±7.567(F=237.182,P0.05),呈浓度依赖性;3.划痕实验结果显示不同浓度的塞来昔布作用24h后PANC-1细胞的迁移能力下降,相对迁移距离分别为11.400±0.959、9.511±1.110、5.856±0.778和2.844±0.416(F=59.516,P0.05),呈浓度依赖性;4.正常PANC-1细胞呈梭形或多角形,状态良好,细胞形态学提示不同浓度的塞来昔布作用24h后PANC-1细胞逐渐变圆、皱缩并开始出现死细胞;5.ELISA结果提示不同浓度的塞来昔布作用24h后胰腺癌细胞中COX-2和MMP-14的蛋白表达水平相应降低,均呈浓度依赖性,COX-2蛋白表达水平分别为0.876±0.023、0.682±0.027、0.505±0.022和0.384±0.019(F=262.537,P0.05);MMP-14蛋白表达水平分别为7.955±0.297、7.031±0.178、6.072±0.209、和5.047±0.216(F=89.247,P0.05);6.PANC-1细胞内COX-2和MMP-14的蛋白表达具有显著正相关性(r=0.873,P0.05)。结论:1.COX-2抑制剂塞来昔布以浓度梯度、时间梯度的形式减弱人胰腺癌细胞系PANC-1的增殖能力,以浓度梯度的形式减弱人胰腺癌细胞系PANC-1的侵袭及迁移能力;2.COX-2抑制剂塞来昔布抑制COX-2表达后,可能通过下调MMP-14表达,进而以浓度梯度形式减弱胰腺癌细胞的增殖、侵袭、迁移能力,起到抗胰腺癌作用。
[Abstract]:Objective: to determine the proliferation, invasion and migration of pancreatic cancer cells by celecoxib, a cyclooxygenase-2 inhibitor with different concentrations of cyclooxygenase-2 (Cyclooxygenase-2). Metalloproteinase 14 matrix metalloproteinase14. To investigate the effect of COX-2 inhibitor celecoxib on pancreatic cancer and its mechanism. Methods: 1. Human pancreatic cancer cell line PANC-1 was cultured in vitro. 2. Four celecoxib concentrations of 0 (0 ~ 0 ~ 20) (0 渭 mol/L) were used to treat the PANC-1 cells at different time intervals (n = 10). The concentration of celecoxib in PANC-1 cells was 100 渭 mol/L (high). 48 and 72 h) MTT colorimetric assay was used to detect the proliferation ability of the cells. Transwell invasion assay and scratch assay were used to detect the invasion ability and migration ability of the cells. The protein expression of COX-2 and MMP-14 was detected by Elisa. 3.The data were statistically analyzed. Results 1. The cell proliferation ability of PANC-1 cells was decreased after 24 h and 72 h treatment with different concentrations of celecoxib. With the increase of celecoxib concentration, the inhibitory effect was gradually enhanced, and with the prolongation of celecoxib action time, the inhibitory effect was gradually strengthened, and the inhibitory effect was time-and concentration-dependent (P0.05). 2. The results of transwell invasion experiment showed that the invasion ability of PANC-1 cells decreased after 24 h treatment with different concentrations of celecoxib. The number of cells passing through the matrix glue was 300.654 卤12.558U 271.041 卤10.569, respectively. 184.003 卤10.865 and 92.667 卤7.567 卤7.567 卤7.567FU 237.182P0.05, in a concentration-dependent manner. 3. The results of scratch test showed that the migration ability of PANC-1 cells was decreased after 24 h treatment with different concentrations of celecoxib, and the relative migration distance was 11.400 卤0.959, respectively. 9.511 卤1.110U 5.856 卤0.778 and 2.844 卤0.416FU 59.516P0.05U in a concentration-dependent manner. 4. The normal PANC-1 cells were fusiform or polygonal, and the morphology indicated that the PANC-1 cells gradually became round after 24 hours of celecoxib treatment with different concentrations of celecoxib. Shrink and begin to appear dead cells; 5. The results of Elisa showed that the protein expression of COX-2 and MMP-14 in pancreatic cancer cells decreased in a concentration dependent manner after 24 hours of treatment with different concentrations of celecoxib. The expression level of COX-2 protein was 0.876 卤0.023 卤0.682 卤0.027, respectively. 0.505 卤0.022 and 0.384 卤0.019 respectively. The expression level of MMP-14 protein was 7.955 卤0.297 卤7.031 卤0.178 卤6.072 卤0.209, respectively. And 5.047 卤0.216F ~ (89) 247 (P _ (0.05)); 6. The expression of COX-2 and MMP-14 in PANC-1 cells was positively correlated with the expression of MMP-14. Conclusion 1. Cyclooxygenase-2 inhibitor celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in the form of concentration gradient and time gradient. The invasion and migration of human pancreatic cancer cell line PANC-1 were attenuated in the form of concentration gradient. 2. COX-2 inhibitor celecoxib inhibits the expression of COX-2, which may decrease the proliferation, invasion and migration of pancreatic cancer cells by down-regulating the expression of MMP-14, and then decreasing the proliferation, invasion and migration of pancreatic cancer cells in the form of concentration gradient. Play an anti-pancreatic cancer effect.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.9

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