MicroRNA-217表达水平与胃癌肿瘤生物学特性分析

发布时间：2018-11-19 21:44

【摘要】：目前,胃癌在因癌症引发的死亡率中排名第二,在男性中的死亡率仅次于支气管肺癌,在女性中的死亡率排名第四。近三分之二的胃癌病例发生在发展中国家,42%发生在中国。胃癌的发生是多因素作用下的多阶段过程,例如癌基因激活、抑制基因失活,免疫改变、表观遗传改变等。即使在接受根治性手术和辅助化疗之后,晚期胃癌患者的病情仍常有恶化,导致治疗失败和高死亡率。因此,急需发现可无创检测出早期胃癌患者的生物标志物。MicroRNAs (miRNA)是～22个核苷酸组成的非编码RNAs,其可参与转录后调控且调控着多种重要的病理生理过程。MiRNA可在多种生物过程的调节中发挥重要作用,包括细胞增殖、凋亡、迁移和分化。异常表达的miRNA与多种疾病有关,包括癌症。既往研究显示,miRNA在胃癌中可扮演抑癌基因或致癌基因的角色。截至目前,有关microRNA-217参与多种肿瘤发生发展的报道很多,但是有关microRNA-217在胃癌中的研究甚少。[目的]探讨microRNA-217在胃癌中的表达水平及其对胃癌发生发展的影响。[方法](1)采用real-time PCR检测50例胃癌组织和癌旁正常组织microRNA-217的表达水平,并进一步分析microRNA-217表达水平与临床病理特征之间的相关性。(2)采用Northern杂交检测胃癌上皮细胞株(SGC-7901、HGC-27、MGC-803、MKN-45)和正常胃黏膜永生化细胞株(GES-1)中microRNA-217的表达水平。(3)采用 real-time PCR检测胃癌上皮细胞株(SGC-7901、HGC-27、MGC-803、MKN-45)和正常胃黏膜永生化细胞株(GES-1)中microRNA-217的表达水平。(4)用microRNA-217模拟物和microRNA-217抑制剂分别转染胃癌上皮细胞株HGC-27后,使用real-timePCR检测其表达水平,表达成功后使用CCK-8试剂盒检测microRNA-217对细胞增殖的影响。(5)应用细胞迁移和侵袭实验技术评估microRNA-217对细胞转移和侵袭能力的影响。[结果](1)相较于癌旁正常组织,microRNA-217的表达水平在胃癌组织中明显下调;并且microRNA-217的表达水平与TNM分期有关。(2) Northern杂交检测结果显示,microRNA-217在胃癌上皮细胞株(SGC-7901、HGC-27、MGC-803、MKN-45)中的表达水平明显低于正常胃黏膜永生化细胞株(GES-1)中的表达水平。(3) Real-time PCR 检测结果显示,microRNA-217 在胃癌细胞株(SGC-7901、HGC-27、MGC-803、MKN-45)中的表达水平明显低于正常胃黏膜永生化细胞株(GES-1)中的表达水平。(4)在胃癌细胞株HGC-27中转染microRNA-217 mimic后,其增殖率显著降低;在胃癌细胞株HGC-27中转染microRNA-217 inhibitor后,其增殖率显著增加。(5)在胃癌细胞株HGC-27中转染microRNA-217 mimic后,其侵袭和转移能力显著降低;在胃癌细胞株HGC-27中转染microRNA-217 inhibitor后,其侵袭和转移能力显著增加。[结论](1) MicroRNA-217在胃癌组织和细胞中的表达水平明显低于癌旁正常组织和细胞中的表达水平。(2) MicroRNA-217表达水平与TNM分期呈负相关。(3)过表达microRNA-217可抑制胃癌细胞增殖、侵袭和转移的能力。
[Abstract]:Gastric cancer is now the second leading cause of cancer mortality among men, second only to lung cancer, and fourth among women. Nearly 2/3 of gastric cancer cases occur in developing countries and 42% in China. The occurrence of gastric cancer is a multistage process under the action of many factors, such as oncogene activation, inhibition gene inactivation, immune change, epigenetic change and so on. Even after radical surgery and adjuvant chemotherapy, advanced gastric cancer patients often deteriorate, leading to treatment failure and high mortality. Therefore, it is urgent to find the noninvasive biomarker. MicroRNAs (miRNA) is a non-coding RNAs, composed of ~ 22 nucleotides in patients with early gastric cancer. MiRNA may play an important role in the regulation of many biological processes, including cell proliferation, apoptosis, migration and differentiation. Abnormally expressed miRNA is associated with many diseases, including cancer. Previous studies have shown that miRNA may play a role as a tumor suppressor gene or oncogene in gastric cancer. Up to now, there have been many reports about the involvement of microRNA-217 in many kinds of tumorigenesis and development, but there are few studies on microRNA-217 in gastric cancer. [objective] to investigate the expression of microRNA-217 in gastric cancer and its influence on the development of gastric cancer. [methods] (1) real-time PCR was used to detect the expression of microRNA-217 in 50 cases of gastric cancer and adjacent normal tissues. Furthermore, the correlation between the expression of microRNA-217 and clinicopathological features was analyzed. (2) Northern hybridization was used to detect the expression of SGC-7901,HGC-27,MGC-803, in gastric cancer epithelial cell line (SGC-7901,HGC-27,MGC-803,). (3) the expression of microRNA-217 was detected by real-time PCR in gastric cancer epithelial cell line (SGC-7901,HGC-27,MGC-803,) and normal gastric mucosa immortalized cell line (GES-1). (4) the expression level of microRNA-217 was detected by real-timePCR after transfection of microRNA-217 mimics and microRNA-217 inhibitor into HGC-27, respectively, in normal gastric mucosa immortalized cell line (GES-1) and normal gastric mucosa immortalized cell line (GES-1). CCK-8 kit was used to detect the effect of microRNA-217 on cell proliferation. (5) the effect of microRNA-217 on cell metastasis and invasion was evaluated by cell migration and invasion assay. [results] (1) the expression of microRNA-217 was down-regulated in gastric cancer tissues compared with the adjacent normal tissues. The expression of microRNA-217 was related to the TNM stage. (2) the results of Northern hybridization showed that the expression of microRNA-217 in gastric cancer epithelial cell line (SGC-7901,HGC-27,MGC-803,) (3) the expression of microRNA-217 in gastric cancer cell line (SGC-7901,HGC-27,MGC-803,) was significantly lower than that in normal gastric mucosa immortalized cell line (GES-1). (3) the results of Real-time PCR showed that the expression of microRNA-217 in gastric cancer cell line (SGC-7901,HGC-27,MGC-803,) was significantly lower than that in normal gastric mucosa immortalized cell line (GES-1). The expression level in MKN-45 was significantly lower than that in normal gastric immortalized cell line (GES-1). (4) after transfection of microRNA-217 mimic into gastric cancer cell line HGC-27, the proliferation rate decreased significantly. After transfection of microRNA-217 inhibitor into gastric cancer cell line HGC-27, the proliferation rate increased significantly. (5) after transfection of microRNA-217 mimic into gastric cancer cell line HGC-27, the ability of invasion and metastasis was significantly decreased. After transfection of microRNA-217 inhibitor into gastric cancer cell line HGC-27, the ability of invasion and metastasis was significantly increased. [conclusion] (1) the expression of MicroRNA-217 in gastric cancer tissues and cells was significantly lower than that in adjacent normal tissues and cells. (2) the expression of MicroRNA-217 was negatively correlated with TNM staging. -217 can inhibit the proliferation of gastric cancer cells, Ability to invade and metastasize.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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