【摘要】：背景腫瘤細胞耐藥是導致腫瘤化學治療失敗的重要原因,長春新堿(vincristine, VCR)是一類廣泛用于腫瘤治療的化學藥物。但是,長期使用可使腫瘤細胞對VCR產(chǎn)生耐藥性,腫瘤耐藥機制仍不清楚。本研究探討與結腸癌耐藥的微小RNA(microRNA, miRNA),旨在揭示腫瘤耐藥機制及為腫瘤細胞耐藥逆轉(zhuǎn)提供基礎。目的建立長春新堿耐藥結腸癌細胞模型,篩選與結腸癌細胞VCR耐藥相關的miRNA。方法1.培養(yǎng)結腸癌HCT-8細胞,采用VCR梯度誘導,建立HCT-8/VCR耐藥細胞株。2.采用全基因組篩選的研究策略,運用HiSeq 2500測序技術和生物信息學的方法,檢測和分析耐藥HCT-8/VCR細胞的和非耐藥的HCT-8細胞差異性表達的miRNAs,構建其miRNA的差異表達譜。3.針對鑒定的差異miRNA,通過靶基因預測軟件進行靶位點確認,通過靶基因所在基因的GO進行功能注釋。結果1.利用濃度梯度的VCR建立了VCR耐藥的結腸癌細胞HCT-8/VCR, HCT-8/VCR細胞在2×103ng/mLVCR中生長良好。2.共進行了1651種miRNA測序,其中有差異的有48種miRNA,與非耐藥的結腸癌HCT-8細胞相比,24種miRNA差異具有統(tǒng)計學意義(P0.05),其中有17種miRNA表達上調(diào)(P0.05)。hsa-miR-675-3p表達量上調(diào)最高,高達7.497681倍,其次是hsa-miR-100-3p, hsa-miR-125b-1-3p, hsa-miR-582-5p, hsa-miR-3141, chr15_10055, hsa-miR-582-5p, hsa-miR-582-3p, hsa-miR-3687。7種miRNA表達下調(diào)(P0.05),下調(diào)最高的是hsa-miR-146a-5p,下調(diào)7.75199倍,其次是hsa-miR-1247-3p, hsa-miR-181 a-3p, hsa-miR-1247-5p。結論]miRNA的異常表達與結腸癌細胞VCR耐藥相關。miRNA表達量的差異可能在結腸癌細胞VCR耐藥中起關鍵作用。
[Abstract]:Background Drug resistance of tumor cells is an important factor leading to the failure of tumor chemotherapy. Vincristine (vincristine, VCR) is a kind of chemotherapeutic drugs widely used in tumor therapy. However, long-term use can result in drug resistance of tumor cells to VCR, and the mechanism of drug-resistance remains unclear. The purpose of this study was to explore the mechanism of drug resistance and to provide a basis for reversal of drug resistance in cancer cells. Objective to establish vincristine resistant colon cancer cell model and screen miRNA. associated with VCR resistance of colon cancer cells. Method 1. Colon cancer HCT-8 cells were cultured and induced by VCR gradient to establish HCT-8/VCR resistant cell line. 2. By using HiSeq 2500 sequencing technique and bioinformatics method, the differentially expressed miRNAs, of drug-resistant HCT-8/VCR cells and non-drug-resistant HCT-8 cells were detected and analyzed by using the strategy of genome screening. The differential expression profile of miRNA was constructed. The target site was identified by target gene prediction software and the functional annotation was performed by GO of the gene in which the target gene was located. Result 1. The VCR resistant colon cancer cell HCT-8/VCR, HCT-8/VCR cells were established by using concentration gradient VCR. The cells grew well in 2 脳 103ng/mLVCR. A total of 1651 miRNA sequences were performed. Among them, 48 miRNA, were significantly different from non-drug-resistant colon cancer HCT-8 cells in 24 miRNA (P0.05), 17 of which were miRNA up-regulated (P0.05), the highest (7.497681 times) in hsa-miR-675-3p expression, and the highest in Hsa-miR-675-3p expression (P0.05). The miRNA expression of hsa-miR-100-3p, hsa-miR-125b-1-3p, hsa-miR-582-5p, hsa-miR-3141, chr15_10055, hsa-miR-582-5p, hsa-miR-582-3p, hsa-miR-3687.7 species was down-regulated (P0.05), and the highest down-regulation of hsa-miR-146a-5p, was 7.75199 times, followed by hsa-miR-1247-3p, hsa-miR-181 a-3p, hsa-miR-1247-5p.. Conclusion: the difference of the expression of miRNA and VCR related to drug resistance in colon cancer cells may play a key role in the drug resistance of colon cancer cell line VCR.
【學位授予單位】：新鄉(xiāng)醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.35

【相似文獻】

相關期刊論文 前10條

1 胡平,琚立華,徐元鼎;結腸癌細胞核DNA含量測量及形態(tài)分析[J];復旦學報(醫(yī)學版);2003年05期

2 王春暉;歐陽欽;唐承薇;黃明慧;李獻;;選擇性及非選擇性COX-2抑制劑對結腸癌細胞生長的影響[J];四川大學學報(醫(yī)學版);2006年04期

3 韓忠寶;李洪祥;楊翊研;;生長抑素聯(lián)合阿霉素體外抑制結腸癌細胞的實驗研究[J];吉林醫(yī)學;2006年09期

4 張朝陽;徐克森;楊廣運;王金申;高穎;劉炎峰;帥晶;王家勇;李少燕;壽楠海;牛軍;;反義整合素β6基因?qū)Y腸癌細胞作用的研究[J];中華醫(yī)學雜志;2007年37期

5 沈丹;鄧長生;;過氧化物酶體增殖物激活受體γ在結腸癌中的表達及其激動劑對結腸癌細胞的生長抑制作用[J];臨床內(nèi)科雜志;2008年01期

6 田永剛;許軍;劉昶;;人工氣腹對結腸癌細胞生物學行為的影響[J];腹腔鏡外科雜志;2009年10期

7 高喜廉，傅志民，姜宗源;結腸癌細胞腸壁縱向浸潤與臨床意義[J];中國醫(yī)科大學學報;1994年03期

8 鄭平菊,黃威權,王瑞安,孫嵐,高平;5-羥色胺及其受體在結腸癌細胞的免疫組織化學定位[J];科學通報;1995年21期

9 林潔;黃瓊;來茂德;;5-氮-2′-脫氧胞苷對結腸癌細胞生物學行為的影響[J];臨床與實驗病理學雜志;2008年04期

10 王德力;齊東麗;;光鑷拉曼光譜技術檢測結腸癌細胞的研究[J];長春大學學報;2009年04期

相關會議論文 前10條

1 許峰;王杉;葉穎江;崔志榮;;腫瘤浸潤淋巴細胞在結腸癌細胞殺傷中的作用及機制[A];中華醫(yī)學會第七次全國消化病學術會議論文匯編（下冊）[C];2007年

2 王貴玉;王錫山;;激光捕獲微切割技術分離結腸癌細胞用于蛋白組學研究[A];中國蛋白質(zhì)組學第三屆學術大會論文摘要[C];2005年

3 沈丹;鄧長生;;過氧化物酶體增殖物激活受體γ在結腸癌中的表達及其激動劑對結腸癌細胞的生長抑制作用[A];中華醫(yī)學會第七次全國消化病學術會議論文匯編（下冊）[C];2007年

4 郭俊明;賴依峰;肖丙秀;劉瓊;;大豆異黃酮對體外培養(yǎng)的結腸癌細胞生長和細胞周期的影響[A];華東六省一市生物化學與分子生物學會2003年學術交流會論文摘要集[C];2003年

5 李本輝;楊賢子;張文杰;;人類結腸癌細胞缺陷型Stat6~(null)活化表型與癌細胞高凋亡率及低侵襲轉(zhuǎn)移力相關[A];湖北省抗癌協(xié)會青年委員會成立大會暨第一屆青年學術論壇資料匯編[C];2009年

6 王國強;方m錁，

本文編號：2215614