辛二酰苯胺异羟肟酸诱导人卵巢癌细胞OC3自噬作用

发布时间：2018-04-16 14:08

本文选题：辛二酰苯胺异羟肟酸 + 卵巢肿瘤　；参考：《中国药理学与毒理学杂志》2015年04期

【摘要】：目的探讨辛二酰苯胺异羟肟酸(SAHA)是否诱导人卵巢癌细胞OC3发生自噬。方法 SAHA0.05,0.25,1.25,6.25和31.25μmol·L-1处理OC3细胞24 h后,倒置显微镜吉姆萨和-瑞氏染色观察细胞形态变化;四甲基偶氮唑蓝(MTT)法检测细胞存活;流式细胞术检测自噬微管相关蛋白轻链3B(LC3B)的表达;AO/EB双染色观察红色酸性自噬囊泡的出现;电镜直接观察自噬囊泡的结构。结果不同浓度SAHA处理后,OC3细胞形态呈不规则梭形或多角形,细胞空泡化增多,折光性差。MTT结果表明,不同浓度SAHA处理对OC3细胞的存活具有明显的抑制作用,并存在时间和浓度效应关系,浓度相关系数分别为r12 h=0.898,r24 h=0.976和r48 h=0.952(P0.05),SAHA 0.05,0.25,1.25,6.25和31.25μmol·L-1时,时间相关系数分别为0.999,0.654,0.999,0.869和0.922(P0.05)。SAHA与OC3细胞作用24 h后,AO/EB双染色可见红色酸性自噬囊泡出现;进一步电镜观察,可观察到自噬囊泡结构;流式细胞术检测结果表明,与正常对照组相比,SAHA 0.05,0.25,1.25,6.25和31.25μmol·L-1处理OC3细胞24 h后,LC3B阳性细胞比率显著升高,分别为(19.4±2.4)%,(28.5±3.4)%,(34.6±3.9)%,(38.6±3.2)%和(61.8±1.0)%(P0.05)。结论SAHA可能通过诱导人卵巢癌细胞OC3发生自噬而杀伤肿瘤细胞。
[Abstract]:Objective to investigate whether octylaniline isoxime acid (SAHAA) induces autophagy in human ovarian cancer cell line OC3.Methods the OC3 cells were treated with SAHA 0.05 0.251.256.25 and 31.25 渭 mol L-1 for 24 h. The morphological changes of OC3 cells were observed by inverted microscope Gimsa and Rayleigh staining, and the survival of OC3 cells was detected by tetramethylazolium.The expression of microtubule-associated protein 3BnLC3B) was detected by flow cytometry. The appearance of red acidic autophagy vesicles was observed by AO-EB double staining, and the structure of autophagy vesicles was observed directly by electron microscope.Results the morphology of OC3 cells was irregular fusiform or polygonal after treatment with different concentrations of SAHA, the vacuolation of cells increased, and the refractive index was poor. The results showed that the survival of OC3 cells was significantly inhibited by different concentrations of SAHA.There was a time-dependent relationship between time and concentration, and the correlation coefficients were r 12 h (r 12 h), 0.898 h (r 24 h) 0.976 and r 48 h (r 48 h), respectively. The time correlation coefficients were 0.9990.6540.9990.869 and 0.9990.6540.9990.869, respectively. When the time correlation coefficients were 0.9990.6540.9990.869 and 0.922(P0.05).SAHA / OC3 cells treated for 24 h, the presence of red acid-macrophage vesicles could be seen by AOEB double staining.Furthermore, the structure of autophagy vesicles was observed by electron microscopy, and the results of flow cytometry showed that compared with the normal control group, the positive rate of OC3 cells was significantly increased after treatment with SAHA0.050.251.256.25 and 31.25 渭 mol L-1 for 24 h, which was 28.5 卤3.44.64.6 卤3.91and 61.8 卤1.0mg / L, respectively.Conclusion SAHA may kill human ovarian cancer cells by inducing OC3 autophagy.
【作者单位】：河北北方学院研究生部;大兴妇幼保健院;空军总医院妇产科;军事医学科学院基础医学研究所细胞生物学研究室;
【基金】：国家自然科学基金(31070996);国家自然科学基金(31171084)~~
【分类号】：R96

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