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背景与目的:染色质核心组蛋白的乙酰化水平受组蛋白乙酰转移酶和组蛋白去乙酰酶(HDAC)的控制,它与基因表达调控密切相关、HDACs功能异常与肿瘤的发生发展有关,组蛋白去乙酰酶抑制剂(HDAC Is)通过抑制HDACs而抗肿瘤。本文研究HDAC I类药物之一MS-275对人膀胱癌T24细胞的抑制作用,为临床应用提供有价值的实验依据。方法:将不同浓度的MS-275作用于T24细胞,用MTT法检测生长抑制作用,用平板克隆形成试验测定细胞增殖能力的影响,分别采用光学显微镜、透射电子显微镜及AnnexinV、PI双标流式细胞术观察和检测细胞的凋亡情况。结果:MS-275对人膀胱癌T24细胞的生长抑制作用存在明显的剂量、时间依赖关系,0.5μmol/L作用72 h的抑制率为9.34%±3.57%,而8μmol/L作用120 h达94.84%±1.33%。克隆形成率从阴性对照组的55.67%±6.51%降至4μmol/L组的2.33%±0.58%。显微镜下可观察到大量的凋亡形态特征的细胞。流式细胞术检测示,作用48 h后的凋亡率在1μmol/L组、2μmol/L组和4μmol/L组分别为24.19%、30.77%和38.51%,明显高于阴性对照组(2.49%)。结论:组蛋白去乙酰酶抑制剂类药物具有体外抗膀胱癌作用,其重要作用机制之一是诱导膀胱癌细胞凋亡,并有望成为新的抗膀胱肿瘤药物。
BACKGROUND & OBJECTIVE: The acetylation level of chromatin core histones is controlled by histone acetyltransferase and histone deacetylase (HDAC), which is closely related to the regulation of gene expression. The dysfunction of HDACs is associated with the development and progression of tumors. Protein deacetylase inhibitors (HDAC Is) are anti-tumor by inhibiting HDACs. This study was to investigate the inhibitory effect of MS-275, a HDAC class I drug, on human bladder cancer T24 cells and provide valuable experimental evidence for clinical application. METHODS: T24 cells were treated with different concentrations of MS-275. The growth inhibition was detected by MTT assay. The proliferation of cells was measured by plate clone formation assay. The cells were stained with optical microscope, transmission electron microscope and Annexin V, Cell observation and detection of cell apoptosis. Results: The inhibitory effect of MS-275 on human bladder cancer T24 cells in a dose-dependent and time-dependent manner was 9.34% ± 3.57% at 0.5 μmol / L for 72 h, and 94.84 at 8 μmol / L for 120 h % ± 1.33%. The rate of colony formation decreased from 55.67% ± 6.51% in the negative control group to 2.33% ± 0.58% in the 4 μmol / L group. A large number of cells characterized by apoptosis were observed under the microscope. Flow cytometry showed that the apoptotic rate of 48 h after treatment was 24.19%, 30.77% and 38.51% in 1μmol / L group, 2μmol / L group and 4μmol / L group respectively, which was significantly higher than that in negative control group (2.49% ). Conclusion: Histone deacetylase inhibitors have the anti-bladder cancer effect in vitro. One of the important mechanisms is to induce apoptosis of bladder cancer cells and to be a new anti-bladder cancer drug.