胃腺癌中LPA差异表达及姜黄素对胃腺癌细胞的影响

发布时间：2018-05-24 06:51

  本文选题：LPA1-3 + 姜黄素　； 参考：《兰州大学》2017年硕士论文

【摘要】：目的:1.探究正常胃黏膜上皮组织、高分化胃腺癌、中分化胃腺癌、低分化胃腺癌中LPA1-3的表达差异水平。2.探究姜黄素(curcumin)对正常胃上皮细胞(GES-1)、高分化胃腺癌细胞(MKN28)、中分化胃腺癌细胞(SGC-7901)及低分化胃腺癌细胞(MKN45)体外增殖的抑制,细胞凋亡的影响以及迁移侵袭能力的影响。方法:1.使用免疫组织化学染色S-P法观察LPA1-3在正常胃上皮及高中低分化胃腺癌组织中表达水平的差异。2.体外培养正常胃黏膜上皮细胞(GES-1)、高分化胃腺癌细胞(MKN28)、中分化胃腺癌细胞(SGC-7901)及低分化胃腺癌细胞(MKN45)。3.Western Blot检测细胞株中LPA1-3表达水平。4.MTT比色法检测姜黄素对四种细胞体外增殖的抑制。5.Hoechst33342染色观察细胞凋亡。6.Transwell小室观察姜黄素对四种细胞迁移侵袭能力的影响。7.应用Imagepro Plus 6.0软件对免疫组织化学染色结果进行分析,所有数据使用SPSS20.0进行分析。结果:1.免疫组化染色结果显示:(1).正常组、高中低分化胃腺癌组中LPA2表达水平均高于LPA1与LPA3(p≤0.05),LPA1与LPA3表达无显著差异(p0.05);(2).LPA1在正常组、高中低胃腺癌组中表达无显著差异(p0.05);LPA2在中分化组中表达低于正常组、高分化组及低分化组(p≤0.05);LPA3在高中低胃腺癌组中表达均低于正常组(p≤0.05),但在高中低胃腺癌组中表达无显著差异(p0.05)。2.各浓度的姜黄素均可抑制GES-1、MKN28、SGC-7901、MKN45四种细胞的增殖,与对照组对比均有显著差异(p≤0.05),且具有时间剂量依赖性。3.姜黄素可诱导GES-1、MKN28、SGC-7901、MKN45四种细胞凋亡的发生。4.姜黄素在低浓度(20μmol/L和40μmol/L)时可促进GES-1细胞的迁移及侵袭能力,在高浓度(80μmol/L)时抑制;姜黄素可抑制MKN28、SGC-7901、MKN45三种细胞迁移及侵袭能力,且呈剂量依赖性。5.Western Blot结果显示:在GES-1细胞中,LPA1高于LPA2和LPA3(p≤0.05),MKN28和SGC-7901细胞中,LPA2均高于LPA1与LPA3(p≤0.05),MKN45细胞中,LPA3高于LPA1与LPA2(p≤0.05)。结论:1.LPA2在正常胃上皮组织、高、中、低分化胃腺癌组织中的表达均高于LPA1与LPA3,在α=0.05水平具有显著差异。2.姜黄素可抑制GES-1、MKN28、SGC-7901、MKN45四种细胞的增殖,并诱导凋亡的发生。3.低浓度姜黄素会促进GES-1细胞迁移及侵袭,在高浓度时抑制;姜黄素对高中低分化胃腺癌细胞迁移及侵袭能力有抑制作用。
[Abstract]:Purpose 1. To investigate the differential expression of LPA1-3 in normal gastric mucosal epithelium, well differentiated gastric adenocarcinoma, moderately differentiated gastric adenocarcinoma and poorly differentiated gastric adenocarcinoma. To investigate the inhibitory effect of curcumin on proliferation, apoptosis and migration and invasion of normal gastric epithelial cells, well-differentiated gastric adenocarcinoma cell line MKN28, moderately differentiated gastric adenocarcinoma cell line SGC-7901) and poorly differentiated gastric adenocarcinoma cell line MKN45) in vitro. Method 1: 1. Immunohistochemical staining S-P method was used to observe the expression of LPA1-3 in normal gastric epithelium and poorly differentiated gastric adenocarcinoma. Normal gastric mucosal epithelial cells were cultured in vitro, well differentiated gastric adenocarcinoma cell line MKN28, moderately differentiated gastric adenocarcinoma cell line SGC-7901) and poorly differentiated gastric adenocarcinoma cell line MKN45. 3. Western Blot were used to detect the expression level of LPA1-3 in cell line. 4. MTT colorimetric assay was used to detect curcumin on four kinds of cells in vitro. Cell apoptosis was observed by Hoechst33342 staining. 6. Transwell chamber was used to observe the effect of curcumin on the migration and invasion of four kinds of cells. The results of immunohistochemical staining were analyzed by Imagepro Plus 6.0 software, and all data were analyzed by SPSS20.0. The result is 1: 1. The results of immunohistochemical staining showed that: 1. There was no significant difference in the expression of LPA2 between LPA1 and LPA3(p 鈮，

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