阿司匹林对人结肠癌HT-29细胞肠分化标志物表达的影响

发布时间：2018-08-21 20:35

【摘要】：背景与目的:非甾体抗炎药阿司匹林临床防治结肠癌确切有效,近年来有研究认为其防治结肠癌最终通过对结肠癌干细胞调控而实现。当结肠癌干细胞分化成熟时,结肠癌将不再进展、甚至消退。本研究旨在探讨非甾体抗炎药阿司匹林对人结肠癌HT-29细胞株分化的影响。方法:采用活细胞计数试剂盒CCK-8检测不同浓度阿司匹林抑制HT-29细胞增殖的作用,得出IC50;采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)检测阿司匹林IC50干预24、48和72 h,肠分化标志物黏蛋白2(mucin 2,MUC2)、三叶因子3(trefoil factor 3,TFF3)、蔗糖酶-异麦芽糖酶(sucroseisomaltase)和溶菌酶(lysozyme)的m RNA表达情况;采用免疫细胞化学法检测MUC2蛋白在细胞中的表达情况;采用蛋白[质]印迹法(Western blot)检测sucrase-isomaltase及lysozyme的蛋白表达情况。结果:阿司匹林能明显抑制HT-29细胞的增殖,且呈时间和剂量依赖性;RTFQ-PCR和Western blot结果显示,阿司匹林干预HT-29细胞48、72 h,与对照组比较,肠杯状细胞标志物MUC2、TFF3的m RNA表达均上调(P0.05),肠吸收细胞标志物sucrase-isomaltase和潘氏细胞标志物lysozyme的m RNA及蛋白表达均下调(P0.05);免疫细胞化学实验结果显示,阿司匹林干预48 h,MUC2蛋白表达较对照组上调,差异有统计学意义(P0.05)。结论:非甾体抗炎药阿司匹林能影响人结肠癌HT-29细胞肠分化标志物的表达,且可能导致其向杯状细胞表型分化。
[Abstract]:Background & AIM: nonsteroidal anti-inflammatory drug aspirin is effective in the prevention and treatment of colon cancer. In recent years, it has been considered that the prevention and treatment of colon cancer can be achieved through the regulation of colon cancer stem cells. When colon cancer stem cells mature, colon cancer will no longer progress, or even fade. The aim of this study was to investigate the effect of aspirin, a non-steroidal anti-inflammatory drug, on the differentiation of human colon cancer cell line HT-29. Methods: the effects of aspirin at different concentrations on the proliferation of HT-29 cells were detected by living cell count kit (CCK-8). The expression of m RNA in intestinal differentiation marker mucin 2 (mucin 2 MUC2), trefoil factor 3 (trefoil factor 3tff3, sucrose enzyme isomaltase (sucroseisomaltase) and lysozyme (lysozyme) were detected by real time real-time fluorescent quantitative polymerase chain reactionation polymerase chain reaction (RTFQ-PCR). The expression of MUC2 protein was detected by immunocytochemistry and the protein expression of sucrase-isomaltase and lysozyme was detected by Western blot (Western blot). Results: aspirin could significantly inhibit the proliferation of HT-29 cells. The results of RT FQ-PCR and Western blot in a time and dose-dependent manner showed that aspirin inhibited the proliferation of HT-29 cells for 4872 hours, compared with the control group. The expression of m RNA in goblet cell marker MUC2TFF3 was up-regulated (P0.05), and the expression of m RNA and protein in intestinal absorption cell marker sucrase-isomaltase and lysozyme were down-regulated (P0.05). The expression of MUC2 protein was up-regulated at 48 h after aspirin intervention compared with the control group (P 0.05). Conclusion: aspirin can affect the expression of intestinal differentiation markers in human colon cancer HT-29 cells and may lead to the phenotype differentiation of goblet cells.
【作者单位】：遵义医学院附属医院消化内科;
【基金】：国家自然科学基金项目(81560467) 贵州省科学技术基金项目(黔科合J字LKZ[2013]18号) 贵州省高层次人才科研条件特助经费资助项目(TZJF-2011-32号) 遵义医学院博士科研启动基金(F-572)
【分类号】：R735.35

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