本文選題：miR-148a-3p + 胃癌��； 參考：《蚌埠醫(yī)學院》2015年碩士論文

【摘要】：背景:胃癌屬于全世界高發(fā)的惡性腫瘤之一,我國胃癌發(fā)病率占惡性腫瘤中占首位,死亡率占所有惡性腫瘤中占第四位,其臨床治療方法通常是手術(shù)切除合并放化療為主。然而,隨著化療在腫瘤病患中的不斷開展,多藥耐藥問題日益凸顯。有研究報道,耐藥是胃癌患者死亡的主要原因。因此,研究胃癌耐藥是醫(yī)務工作人員亟需解決的問題。Micro RNAs是一組含量豐富,長度為20各堿基左右的單鏈非編碼小RNA。它通過與靶基因mRNA的3,-非編碼區(qū)互補配對,直接誘導靶基因mRNA降解或抑制其蛋白質(zhì)合成,從而在細胞分化、凋亡、增值、新陳代謝等生命過程發(fā)揮重要作用。已有大量文獻報道上調(diào)或下調(diào)腫瘤耐藥相關(guān)的miRNA分子可逆轉(zhuǎn)腫瘤耐藥。我們參考國內(nèi)外相關(guān)文獻,得出miR-148a-3p與胃癌耐藥相互關(guān)系尚沒有報道,因此我們展開以下實驗。目的:探究miR-148a-3p調(diào)節(jié)胃癌多重耐藥作用機制方法:1.QRT-PCR檢測比較耐藥細胞系SGC7901/ADR與其親本細胞SGC7901之間miR-148a-3p表達差異。2.采用瞬時轉(zhuǎn)染miR-148a-3p mimic和inhibitor,分別上調(diào)SGC7901/ADR細胞miR-148a-3p表達量,下調(diào)SGC7901細胞miR-148a-3p表達量。通過體外MTT藥敏實驗檢測miR-148a-3p對其細胞耐藥的影響,流式細胞技術(shù)檢測miR-148a-3p對細胞凋亡情況的影響。3.深入研究miR-148a-3p調(diào)節(jié)胃癌多重耐藥可能的機制。結(jié)果:1.耐藥細胞SGC7901/ADR中miR-148a-3p表達量低于其親本細胞SGC7901。2.上調(diào)SGC7901/ADR細胞中miR-148a-3p表達量后,可增加耐藥細胞對化療藥物的敏感性,增加細胞凋亡百分比;下調(diào)SGC7901細胞中miR-148a-3p表達后,可降低SGC7901細胞對化療藥物的敏感性,并減少細胞凋亡百分比。3.Western-blot及q RT-PCR結(jié)果顯示ERBB3可能是miR-148a-3p的靶基因之一。用siRNA降低SGC7901/ADR細胞中ERBB3表達后,其對化療藥物的敏感性增加。4.Western-blot結(jié)果顯示,下調(diào)SGC7901/ADR細胞中ERBB3表達后,p-Akt表達水平降低。結(jié)論:miR-148a-3p可能通過作用于其靶基因ERBB3,介導ERBB3/PI3K/Akt信號通路調(diào)節(jié)胃癌多重耐藥。
[Abstract]:Background: gastric cancer is one of the most common malignant tumors in the world. The incidence of gastric cancer is the first and the mortality rate is the fourth among all malignant tumors in China. The clinical treatment of gastric cancer is usually surgical resection combined with radiotherapy and chemotherapy. However, with the continuous development of chemotherapy in cancer patients, the problem of multi-drug resistance has become increasingly prominent. Drug resistance is reported to be the leading cause of death in patients with gastric cancer. Therefore, the study of drug resistance in gastric cancer is an urgent problem for medical workers. Micro RNAs are a group of single-stranded non-coding small RNAs with rich contents and about 20 bases in length. It directly induces the degradation of target gene mRNA or inhibits its protein synthesis by complementing with the 3- non-coding region of target gene mRNA, thus plays an important role in cell differentiation, apoptosis, proliferation, metabolism and other life processes. A large number of literatures have reported that up-regulating or down-regulating miRNA molecules associated with tumor resistance can reverse drug resistance. The relationship between miR-148a-3p and gastric cancer resistance has not been reported, so we have carried out the following experiments. Aim: to investigate the mechanism of miR-148a-3p in regulating multidrug resistance in gastric cancer cell line SGC7901 / ADR and its parent cell line SGC7901. Methods: 1. The expression of miR-148a-3p in gastric cancer cell line SGC7901 / ADR was compared with that of its parent cell line SGC7901. Transient transfection of miR-148a-3p mimic and inhibitor were used to up-regulate the expression of miR-148a-3p and down-regulate the expression of miR-148a-3p in SGC7901 cells. The effect of miR-148a-3p on cell resistance was detected by MTT assay in vitro, and the effect of miR-148a-3p on cell apoptosis was detected by flow cytometry. To study the possible mechanism of miR-148a-3p regulating multidrug resistance in gastric cancer. The result is 1: 1. The expression of miR-148a-3p in SGC7901 / ADR was lower than that in SGC7901.2. After upregulation of miR-148a-3p expression in SGC7901 / ADR cells, the sensitivity of resistant cells to chemotherapeutic drugs and the percentage of apoptosis were increased, and the sensitivity of SGC7901 cells to chemotherapeutic drugs was decreased after down-regulating the expression of miR-148a-3p in SGC7901 cells. The results of Western-blot and Q RT-PCR showed that ERBB3 might be one of the target genes of miR-148a-3p. The sensitivity of ERBB3 in SGC7901 / ADR cells was increased with siRNA. 4. Western-blot results showed that the expression of ERBB3 decreased after down-regulation of ERBB3 expression in SGC7901 / ADR cells. Conclusion it is possible that the multidrug resistance of gastric cancer is mediated by the ERBB3 / PI3K / Akt signaling pathway mediated by the action of the target gene ERBB3: miR-148a-3p.
【學位授予單位】：蚌埠醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.2

【參考文獻】

相關(guān)期刊論文 前1條

1 Sumadi Lukman Anwar;Ulrich Lehmann;;DNA methylation,microRNAs,and their crosstalk as potential biomarkers in hepatocellular carcinoma[J];World Journal of Gastroenterology;2014年24期

，

本文編號：2086057