發(fā)布時(shí)間：2018-03-29 20:43

  本文選題：胃癌　切入點(diǎn)：奧沙利鉑　出處：《首都醫(yī)科大學(xué)》2016年碩士論文

【摘要】：研究目的：本研究旨在探索miR-145與胃癌奧沙利鉑耐藥的關(guān)系。材料與方法：通過(guò)濃度遞增方法誘導(dǎo)出對(duì)奧沙利鉑耐藥的胃癌細(xì)胞株BGC-823/L-OHP,應(yīng)用MTT法檢測(cè)兩組ICso驗(yàn)證耐藥株誘導(dǎo)成功。通過(guò)實(shí)時(shí)定量PCR方法檢驗(yàn)野生BGC-823及對(duì)奧沙利鉑耐藥株BGC-823/L-OHP兩組細(xì)胞的miR-145表達(dá)水平,驗(yàn)證miR-145與胃癌細(xì)胞奧沙利鉑耐藥相關(guān)性。構(gòu)建差異表達(dá)miR-145的四組BGC-823細(xì)胞系(高表達(dá)組、低表達(dá)組、空載體組、野生組),通過(guò)MTT法檢測(cè)四組細(xì)胞對(duì)奧沙利鉑的半數(shù)抑制濃度(IC50),在細(xì)胞水平進(jìn)一步驗(yàn)證miR-145與胃癌細(xì)胞奧沙利鉑耐藥相關(guān)性。結(jié)果：構(gòu)建耐藥株后,通過(guò)MTT實(shí)驗(yàn)顯示BGC-823細(xì)胞株的半抑制濃度(IC50)為8.164ug/ml, BGC-823/L-OHP細(xì)胞株的ICso為138.994ug/ml,較前者明顯升高,耐藥株建立成功。通過(guò)病毒向野生BGC-823細(xì)胞系中轉(zhuǎn)染靶基因,熒光顯微鏡下可觀察到四組細(xì)胞內(nèi)可見(jiàn)熒光蛋白,成功構(gòu)建四組差異表達(dá)miR-145的胃癌細(xì)胞系(高表達(dá)組、低表達(dá)組、空載體組、野生組),MTT實(shí)驗(yàn)結(jié)果顯示高表達(dá)miR-145組IC50為9.861ug/ml,低表達(dá)miR-145組ICso為5.377ug/ml,空載體組ICso為5.318ug/ml,野生組ICso為4.805ug/ml,高表達(dá)組表現(xiàn)出奧沙利鉑耐藥性升高。結(jié)論：miR-145在BGC-823細(xì)胞株與BGC-823/L-OHP田胞株之間表達(dá)存在差異,耐藥株中表達(dá)明顯升高；高表達(dá)miR-145后,胃癌細(xì)胞系BGC-823對(duì)奧沙利鉑耐藥性升高。miR-145可作為預(yù)測(cè)胃癌奧沙利鉑化療是否敏感的標(biāo)志物,高表達(dá)miR-145的胃癌細(xì)胞奧沙利鉑化療敏感性欠佳。
[Abstract]:Objective: to explore the relationship between miR-145 and oxaliplatin resistance in gastric cancer. Materials and methods: to induce oxaliplatin resistant gastric cancer cell line BGC-823 / L-OHPby increasing concentration, and to detect the drug resistance of two groups by MTT assay. The expression of miR-145 was detected by real-time quantitative PCR assay in both groups of BGC-823 and BGC-823/L-OHP cells, which were resistant to oxaliplatin. To verify the correlation between miR-145 and oxaliplatin resistance in gastric cancer cells, four groups of BGC-823 cell lines (high expression group, low expression group, empty vector group) with differentially expressed miR-145 were constructed. In wild group, the half inhibitory concentration of oxaliplatin on oxaliplatin was detected by MTT method, and the correlation between miR-145 and oxaliplatin resistance in gastric cancer cell line was further verified at the cell level. MTT assay showed that the semi-inhibitory concentration of BGC-823 cell line IC50 was 8.164ugr / ml, and the ICso of BGC-823/L-OHP cell line was 138.994ugr / ml, which was significantly higher than that of the former cell line, and the drug resistant strain was successfully established. The target gene was transfected into the wild BGC-823 cell line by virus transfection. Four groups of gastric cancer cell lines with differentially expressed miR-145 (high expression group, low expression group, empty vector group) were successfully constructed. The results of wild group showed that the IC50 of high expression miR-145 group was 9.861 ugr / ml, the ICso of low expression miR-145 group was 5.377ugr / ml, the ICso of empty vector group was 5.318ugr / ml, the ICso of wild group was 4.805ugr / ml, and the drug resistance of oxaliplatin was increased in high expression group. There are differences in expression between two groups. After high expression of miR-145, the drug resistance of gastric cancer cell line BGC-823 to oxaliplatin was increased. MiR-145 could be used as a marker to predict the chemosensitivity of oxaliplatin to oxaliplatin in gastric cancer, but the chemosensitivity of gastric cancer cell line with high expression of miR-145 to oxaliplatin was not good.
【學(xué)位授予單位】：首都醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前4條

1 張晴;吳正升;張瑰紅;丁向東;黃焰;楊楓;徐曉春;吳強(qiáng);;miR-145在乳腺癌中表達(dá)及意義[J];臨床與實(shí)驗(yàn)病理學(xué)雜志;2009年01期

2 郭廣秀;張功亮;陳青;;miR-145與腫瘤研究進(jìn)展[J];江西醫(yī)藥;2011年11期

3 董文;林天歆;黃健;廖蓓;許可慰;江春;張彩霞;劉皓;;miR-143和miR-145在膀胱癌中的表達(dá)及意義[J];中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版);2011年06期

4 ;[J];;年期

相關(guān)會(huì)議論文 前2條

1 馮陽(yáng);曹建華;李新云;趙書(shū)紅;;miR-145在豬骨骼肌發(fā)育和成肌細(xì)胞分化不同時(shí)期的差異表達(dá)分析[A];中國(guó)遺傳學(xué)會(huì)模式生物與人類健康研討會(huì)會(huì)議論文集[C];2010年

2 馮陽(yáng);曹建華;李新云;趙書(shū)紅;;miR-145在豬骨骼肌發(fā)育和成肌細(xì)胞分化不同時(shí)期的差異表達(dá)分析[A];中國(guó)遺傳學(xué)會(huì)模式生物與人類健康研討會(huì)會(huì)議論文集[C];2010年

相關(guān)博士學(xué)位論文 前1條

1 董瑞芬;miR-145在漿液性卵巢癌發(fā)生發(fā)展中的作用和機(jī)制研究[D];山東大學(xué);2015年

相關(guān)碩士學(xué)位論文 前5條

1 尉建安;胃癌對(duì)奧沙利鉑耐藥性與miR-145關(guān)系初探[D];首都醫(yī)科大學(xué);2016年

2 陳敏;miR-145介導(dǎo)阿司匹林抗血管平滑肌細(xì)胞增殖及抗炎作用的研究[D];中南大學(xué);2014年

3 鄧鑫;miR-145對(duì)前列腺癌PC-3細(xì)胞增殖和凋亡及其調(diào)控基因表達(dá)的影響[D];蘇州大學(xué);2014年

4 陳嘉佳;miR-145靶向FSCN1在不同生長(zhǎng)方式胃癌中的作用及機(jī)制[D];廈門大學(xué);2014年

5 張鴻博;miR-145對(duì)宮頸癌Hela細(xì)胞的增殖、凋亡及其對(duì)紫杉醇藥物敏感性的研究[D];吉林大學(xué);2014年

，

本文編號(hào)：1682766