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(P<0.05),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(P<0.05);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(P<0.05)。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。
：Background and purpose： Aspirin （one of the nonsteroidal anti-inflammatory drugs） is effective in the prevention and treatment of colon cancer. And recently, it has been indicated that regulating colon cancer stem cells might be the underlying mechanism. When colon cancer stem cells become mature, colon cancer will not progress, and even subside. Therefore, this study aimed to explore the effects of aspirin on differentiation in human colon cancer cell line HT-29. Methods： The inhibitory effects of different concentrations of aspirin on HT-29 cells were detected by living cell counting kit CCK-8, and the IC50 was calculated. Real-time fluorescence quantitative polymerase chain reaction （RTFQ-PCR） was used to detect the mRNA expressions of intestinal differentiation markers [mucin 2 （MUC2）, trefoil factor 3 （TFF3） lysozyme and sucrase-isomaltase] after aspirin IC50 intervention for 24, 48 and 72 h. Immunocytochemistry was used to detect the MUC2 protein expression. And Western blot was used to detect the protein expressions of lysozyme and sucrase-isomaltase. Results： Aspirin inhibited the proliferation of HT-29 cells significantly in a time- and dose-dependent manner. RTFQ-PCR and Western blot showed that the mRNA expressions of goblet cell markers （MUC2 and TFF3） were increased （P〈0.05）, and the mRNA and protein expressions of Paneth cell marker （lysozyme） and absorptive cell marker （sucrase-isomaltase） were decreased （P〈0.05）, compared with the control group, after aspirin intervention for 48 and 72 h. Immunocytochemistry showed that the MUC2 protein expression was increased （P〈0.05） compared with the control group after aspirin intervention for 48 h. Conclusion： Nonsteroidal anti-inflammatory drug aspirin can affect the expressions of intestinal differentiation markers in human colon cancer HT-29 cells, and may lead to their differentiation into the phenotype of goblet cell.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2017年第11期867-872,共6页
China Oncology
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)
