探究吉西他滨对膀胱肿瘤T24细胞MicroRNA表达谱的影响

发布时间：2018-05-15 06:15

  本文选题：膀胱癌 + 吉西他滨　； 参考：《蚌埠医学院》2014年硕士论文

【摘要】：研究背景：膀胱癌是泌尿系统中最常见的肿瘤，近年来膀胱癌的发病率有逐渐升高的趋势，已经成为严重威胁人类健康的常见疾病；随着医疗水平的提高，化疗药物的出现明显降底了膀胱癌术后的复发率。吉西他滨是目前临床一线的化疗药物，通过作用于DNA合成期阻止其进展来发挥抗肿瘤的效应，在膀胱肿瘤的治疗中，其疗效已经得到大量临床试验的验证。最近有研究表明吉西他滨能通过微小RNA (miRNA)发挥其抗肿瘤作用；miRNA是一类非编码小分子RNA，通过与靶基因mRNA的不完全配对从而抑制其翻译或直接将其降解；作为重要的基因调控因子发挥着癌基因或抑癌基因的作用。MiRNA与肿瘤发生发展的关系成为近年来的研究热点。虽然在治疗膀胱癌上有一定的疗效，但是药物的作用靶点以及转录后水平的调节机制还尚不清楚；本研究采用基因芯片技术，通过检测吉西他滨干预膀胱肿瘤T-24细胞后miRNA表达谱的差异性来分析其可能抑制膀胱肿瘤细胞生长的机制。 目的：探究吉西他滨对膀胱肿瘤T24细胞miRNA表达谱的影响。 方法：培养膀胱肿瘤T-24细胞株，待其传代生长后用MTT法检测细胞增殖情况并计算吉西他滨的半数抑制浓度（即IC50);用此浓度刺激T-24细胞24h后，利用miRNA芯片技术检测用药前后T-24细胞miRNA的表达，对其表达谱进行差异性分析，并筛选出差异表达的miRNA分子。 结果：1.T-24细胞的生长抑制率与吉西他滨之间存在一定的浓度依赖关系，测出其IC50为1.403ug/ml。 2.用IC50刺激T24细胞24h后，利用miRNA芯片技术检测加药刺激后的样本与未加药的样本比较。共测得有152个与吉西他滨加药刺激相关的miRNA差异表达，其中与未刺激的样本比较明显上调的有7个，包括miR-200c-5p,miR-3689b-3p/miR-3689c，miR-2355-5p，miR-195-3p，miR-23b-5p，miR-4648禾口miR-3929；明显下调的有11个，包括miR-802，miR-1252-5p，miR-5009-3p，miR-5003-5p，miR-874-3p，miR-216a-5p? miR-4736，miR-548aa/miR-548ap-3/miR-548t-3p?miR-548h-3p/miR-548z，miR-3688-5p，，miR-2681-3p。 结论：1.吉西他滨与膀胱肿瘤T-24细胞的增殖呈浓度依赖关系；并且吉西他滨的干预能够使T-24细胞的miRNA表达谱发生改变 2.吉西他滨对miRNA的表达调控可能是其抗膀胱肿瘤T-24细胞的重要途径。
[Abstract]:Background: bladder cancer is the most common tumor in the urinary system. In recent years, the incidence of bladder cancer has gradually increased, which has become a serious threat to human health. The appearance of chemotherapeutic drugs significantly reduced the recurrence rate of bladder cancer after operation. Gemcitabine is one of the first-line chemotherapeutic drugs in clinic at present. It has been proved by a large number of clinical trials that gemcitabine can prevent the progress of DNA in the synthetic phase to play an anti-tumor effect. Recent studies have shown that gemcitabine can play its anti-tumor effect through small RNA miRNAs. It is a kind of non-coding small molecule RNAs, which inhibits its translation or directly degrades by incomplete pairing with the target gene mRNA. As an important gene regulator, the role of oncogene or tumor suppressor gene. MiRNA has become a hot topic in recent years. Although there are some therapeutic effects in the treatment of bladder cancer, the target of the drug and the regulation mechanism of posttranscriptional level are still unclear. By detecting the difference of miRNA expression profile after gemcitabine intervention in bladder tumor T-24 cells, the mechanism of inhibition of bladder tumor cell growth was analyzed. Objective: to investigate the effect of gemcitabine on miRNA expression profile of bladder tumor T 24 cells. Methods: the bladder tumor T-24 cell line was cultured. The proliferation of T-24 cells was detected by MTT method and the half inhibitory concentration (IC50) of gemcitabine was calculated after 24 h stimulation of T-24 cells. The expression of miRNA in T-24 cells before and after treatment was detected by miRNA chip technique. The differential expression profiles were analyzed and the differential expression miRNA molecules were screened out. Results there was a concentration-dependent relationship between the growth inhibition rate and gemcitabine, and the IC50 was 1.403U / ml. 2. After T24 cells were stimulated with IC50 for 24 hours, miRNA microarray technique was used to detect the comparison between the samples stimulated by IC50 and those without. 鍏辨祴寰楁湁152涓�涓庡悏瑗夸粬婊ㄥ姞鑽�鍒烘縺鐩稿叧鐨刴iRNA宸�寮傝〃杈�,鍏朵腑涓庢湭鍒烘縺鐨勬牱鏈�姣旇緝鏄庢樉涓婅皟鐨勬�

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