兰州大学机构库 >学院待认领
COX-2调控胃癌细胞侵袭迁移分子机制的实验研究
Alternative TitleTo Explore the Molecular Mechanism of Invasion and Migration Regulated by Cyclooxygenase-2 in Human Gastric Carcinoma Cell in vitro
刘敏
Thesis Advisor周永宁
2010-05-21
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword环氧化酶-2 基质金属蛋白酶-2 E-钙粘素 前列腺素E2 塞来昔布 侵袭 迁移 胃癌
Abstract胃癌是一种发病率很高的恶性肿瘤,尤其是在亚洲国家,近年来其发病率呈上升趋势。胃癌的早期临床症状不典型,患者就诊时多已经属于晚期并有转移,死亡率高、预后差。如何早期诊断、监测术后复发、远处转移是进一步提高胃癌综合疗效的关键。环氧化酶2(COX-2)是催化花生四烯酸生成前列腺素E2(PGE2)的关键酶。近年来国内外大量的研究表明,COX-2在多种实体肿瘤中过表达,它不仅通过促进细胞增殖、抑制凋亡参与多种肿瘤的发生过程,而且还通过各种机制参与肿瘤的侵袭与转移。迄今为止,COX-2参与胃癌侵袭、转移的机制尚不清楚。本研究通过使用特异性COX-2抑制剂塞来昔布和前列腺素E2对体外培养的人胃癌细胞株SGC-7901进行干预,探讨COX-2对E-cadherin、基质金属蛋白酶2(MMP-2)表达的影响,以及COX-2在胃癌侵袭迁移中的作用,为胃癌的早期诊断和治疗提供有价值的分子生物学指标。目的 使用塞来昔布、PGE2对人胃癌细胞株SGC-7901进行干预,探讨COX-2对E-cadherin、MMP-2表达的影响,阐明COX-2与E-cadherin、MMP-2之间的相互关系及其参与胃癌细胞侵袭迁移的可能机制。方法 使用特异性COX-2抑制剂及PGE2对体外培养的人胃癌细胞系SGC-7901进行干预,采用实时荧光定量反转录PCR分别检测COX-2、E-cadherin、MMP-2 mRNA的表达;运用免疫荧光标记法结合激光共聚焦荧光显微镜分析E-cadherin蛋白表达量;应用Transwell法检测塞来昔布、PGE2对胃癌细胞侵袭及迁移能力的影响。结果 ①实时荧光定量反转录PCR实验:塞来昔布明显抑制体外培养人胃癌细胞SGC-7901中COX-2 mRNA的表达,且E-cadherin mRNA的表达逐渐升高,MMP-2 mRNA的表达逐渐下降;加入PGE2干预后E-cadherin mRNA的表达下降,MMP-2 mRNA的表达增强。②应用塞来昔布作用于人胃癌细胞SGC-7901后,激光共聚焦荧光显微镜检测E-cadherin的蛋白表达量明显升高;使用PGE2处理SGC-7901细胞后其E-cadherin蛋白表达量明显下降。③塞来昔布处理组胃癌细胞穿过Transwell小室的细胞数明显少于对照组,塞来昔布能抑制SGC-7901细胞的侵袭和迁移能力;PGE2处理组细胞穿过Transwell小室的胃癌细胞数明显多于对照组,PGE2能促进SGC-7901细胞的侵袭和迁移。结论 使用塞来昔布干预后能上调SGC-7901细胞中E-cadherin基因及蛋白的表达,下调MMP-2基因的表达,抑制胃癌细胞体外侵袭和迁移。PGE2干预后能下调SGC-7901细胞中E-cadherin基因及蛋白的表达,上调MMP-2基因的表达,促进胃癌细胞体外侵袭和迁移。我们的实验证实COX-2通过调控E-cadherin、MMP-2的表达参与胃癌侵袭迁移的过程。
Other AbstractGastric cancer is a malignant tumor with a high incidence, especially in Asian countries, its incidence showed an upgrade trend in recent years. The early clinical symptom of gastric cancer is not typical, and a lot of patients diagnosed with gastric cancer over time and have been part of the transfer of advanced stage with high mortality and poor prognosis. The key of further improve the comprehensive efficacy of gastric cancer is how to early diagnose and monitor recurrence, distant metastasis after surgery. Cyclooxygenase-2 (COX-2) is the key enzyme which catalyzed arachidonic acid into prostaglandin E2 (PGE2). A large number of studies at home and abroad in recent years have showed that COX-2 is over-expression in a variety of solid tumors, which not only involved in the tumorigenesis and tumor development process according to promote cell proliferation, inhibit of apoptosis, but also involved in tumor invasion and metastasis through various mechanisms. To date, the mechanism of COX-2 involved in gastric cancer invasion and metastasis is not clear. In this study, the specific COX-2 inhibitor celecoxib and prostaglandin E2 have been used to intervene in cultured human gastric cancer cell line SGC-7901 in vitro. We explored the effect of COX-2 on the expression of E-cadherin and matrix metalloproteinase2 (MMP-2), as well as the role of COX-2 in gastric cancer and migration and provide valuable indicators of molecular biology for the early diagnosis and treatment of gastric cancer. Aim:Celecoxib and PGE2 have been used to intervene in human gastric cancer cell line SGC-7901. We explored the effect of COX-2 on the expression of E-cadherin and MMP-2, as well as the relationship between the expression of COX-2 and E-cadherin, MMP-2, and the mechanism of COX-2 involvement in the tumor cell invasion and migration. Method:After celecoxib and PGE2 have been used to intervene in the gastric carcinoma cell line SGC-7901, the real-time quantitative PCR was used to detect the level of COX-2, E-cadherin and MMP-2 mRNA. The combination immunofluorescence and confocal laser scanning microscopy was applied to analyze the expression of E-cadherin protein. Transwell was used to detect the effect of celecoxib and PGE2 on migration and invasive potential of human gastric carcinoma cell. Result:①In the real-time quantitative PCR, celecoxib significantly inhibited the COX-2 mRNA expression, with a corresponding E-cadherin mRNA expression gradually increased and MMP-2 mR...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/224185
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
刘敏. COX-2调控胃癌细胞侵袭迁移分子机制的实验研究[D]. 兰州. 兰州大学,2010.
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