【摘要】：目前,胃癌在因癌癥引發(fā)的死亡率中排名第二,在男性中的死亡率僅次于支氣管肺癌,在女性中的死亡率排名第四。近三分之二的胃癌病例發(fā)生在發(fā)展中國家,42%發(fā)生在中國。胃癌的發(fā)生是多因素作用下的多階段過程,例如癌基因激活、抑制基因失活,免疫改變、表觀遺傳改變等。即使在接受根治性手術(shù)和輔助化療之后,晚期胃癌患者的病情仍常有惡化,導(dǎo)致治療失敗和高死亡率。因此,急需發(fā)現(xiàn)可無創(chuàng)檢測出早期胃癌患者的生物標(biāo)志物。MicroRNAs (miRNA)是～22個(gè)核苷酸組成的非編碼RNAs,其可參與轉(zhuǎn)錄后調(diào)控且調(diào)控著多種重要的病理生理過程。MiRNA可在多種生物過程的調(diào)節(jié)中發(fā)揮重要作用,包括細(xì)胞增殖、凋亡、遷移和分化。異常表達(dá)的miRNA與多種疾病有關(guān),包括癌癥。既往研究顯示,miRNA在胃癌中可扮演抑癌基因或致癌基因的角色。截至目前,有關(guān)microRNA-217參與多種腫瘤發(fā)生發(fā)展的報(bào)道很多,但是有關(guān)microRNA-217在胃癌中的研究甚少。[目的]探討microRNA-217在胃癌中的表達(dá)水平及其對胃癌發(fā)生發(fā)展的影響。[方法](1)采用real-time PCR檢測50例胃癌組織和癌旁正常組織microRNA-217的表達(dá)水平,并進(jìn)一步分析microRNA-217表達(dá)水平與臨床病理特征之間的相關(guān)性。(2)采用Northern雜交檢測胃癌上皮細(xì)胞株(SGC-7901、HGC-27、MGC-803、MKN-45)和正常胃黏膜永生化細(xì)胞株(GES-1)中microRNA-217的表達(dá)水平。(3)采用 real-time PCR檢測胃癌上皮細(xì)胞株(SGC-7901、HGC-27、MGC-803、MKN-45)和正常胃黏膜永生化細(xì)胞株(GES-1)中microRNA-217的表達(dá)水平。(4)用microRNA-217模擬物和microRNA-217抑制劑分別轉(zhuǎn)染胃癌上皮細(xì)胞株HGC-27后,使用real-timePCR檢測其表達(dá)水平,表達(dá)成功后使用CCK-8試劑盒檢測microRNA-217對細(xì)胞增殖的影響。(5)應(yīng)用細(xì)胞遷移和侵襲實(shí)驗(yàn)技術(shù)評(píng)估m(xù)icroRNA-217對細(xì)胞轉(zhuǎn)移和侵襲能力的影響。[結(jié)果](1)相較于癌旁正常組織,microRNA-217的表達(dá)水平在胃癌組織中明顯下調(diào);并且microRNA-217的表達(dá)水平與TNM分期有關(guān)。(2) Northern雜交檢測結(jié)果顯示,microRNA-217在胃癌上皮細(xì)胞株(SGC-7901、HGC-27、MGC-803、MKN-45)中的表達(dá)水平明顯低于正常胃黏膜永生化細(xì)胞株(GES-1)中的表達(dá)水平。(3) Real-time PCR 檢測結(jié)果顯示,microRNA-217 在胃癌細(xì)胞株(SGC-7901、HGC-27、MGC-803、MKN-45)中的表達(dá)水平明顯低于正常胃黏膜永生化細(xì)胞株(GES-1)中的表達(dá)水平。(4)在胃癌細(xì)胞株HGC-27中轉(zhuǎn)染microRNA-217 mimic后,其增殖率顯著降低;在胃癌細(xì)胞株HGC-27中轉(zhuǎn)染microRNA-217 inhibitor后,其增殖率顯著增加。(5)在胃癌細(xì)胞株HGC-27中轉(zhuǎn)染microRNA-217 mimic后,其侵襲和轉(zhuǎn)移能力顯著降低;在胃癌細(xì)胞株HGC-27中轉(zhuǎn)染microRNA-217 inhibitor后,其侵襲和轉(zhuǎn)移能力顯著增加。[結(jié)論](1) MicroRNA-217在胃癌組織和細(xì)胞中的表達(dá)水平明顯低于癌旁正常組織和細(xì)胞中的表達(dá)水平。(2) MicroRNA-217表達(dá)水平與TNM分期呈負(fù)相關(guān)。(3)過表達(dá)microRNA-217可抑制胃癌細(xì)胞增殖、侵襲和轉(zhuǎn)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.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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本文編號(hào)：2343507