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目的研究辛二酞苯胺异羟肟酸(SAHA)对人卵巢癌紫杉醇耐药细胞OC3/P存活和凋亡的影响。方法 SAHA 4~64μmo·lL-1与OC3/P细胞作用6,12,24或48 h,用倒置显微镜观察细胞形态的变化,瑞氏吉姆萨染色观察细胞质和细胞核形态的变化,MTT法检测细胞存活率,AnnexinⅤ-FITC/PI双染流式细胞术测定细胞凋亡率,用透射电镜观察细胞内超微结构的变化。结果倒置显微镜下可见,SAHA 4,16和64μmo·lL-1与OC3/P细胞孵育24 h,可引起OC3/P细胞皱缩,细胞贴壁不良,折光率减弱,活细胞数目减少,且随着SAHA浓度升高细胞形态改变更明显。瑞氏吉姆萨染色发现,SAHA 64μmol·L-1与OC3/P细胞孵育24 h,使细胞体积减小、细胞质凝集和核固缩。MTT法检测结果表明,SAHA 4~64μmo·lL-1对OC3/P细胞存活具有抑制作用,并存在明显的时间和浓度效应关系,其中SAHA 64μmol·L-1作用6,12,24和48 h对OC3/P细胞存活的抑制率分别为(7.1±1.9)%,(14.6±2.0)%,(36.8±2.7)%和(83.3±3.0)%(r=0.891,P<0.05),SAHA 4,8,16,32和64μmo·lL-1作用48 h对OC3/P细胞存活的抑制率分别为(28.8±1.2)%,(34.8±4.3)%,(52.3±2.8)%,(61.3±4.9)%和(83.3±3.0)%(r=0.966,P<0.05)。流式细胞术检测结果表明,SAHA 4,16和64μmo·lL-1作用48 h均可诱导OC3/P细胞凋亡(P<0.05,P<0.01)。透射电镜观察结果表明,SAHA 4,16和64μmo·lL-1作用24 h,OC3/P细胞核出现典型的凋亡形态变化。结论 SAHA可抑制人卵巢癌紫杉醇耐药细胞OC3/P的存活并诱导其凋亡。
Objective To study the effects of saphenanthroline aniline hydroxamic acid (SAHA) on the survival and apoptosis of OC3 / P cells in paclitaxel resistant human ovarian cancer cells. Methods SAHA 4 ~ 64μmo · lL-1 and OC3 / P cells were treated with 6, 12, 24 or 48 hours. The morphology of the cells was observed by inverted microscope. The changes of cytoplasm and nucleus were observed by Wright Giemsa staining. Survival rate, Annexin Ⅴ-FITC / PI double staining flow cytometry was used to determine the apoptotic rate. Transmission electron microscopy was used to observe the changes of intracellular ultrastructure. Results Inverted microscope showed that SAHA 4, 16 and 64μmo · lL-1 incubated with OC3 / P cells for 24 hours caused OC3 / P cells to shrink, poorly adherent cells, weakened refractive index, and decreased number of viable cells SAHA concentration increased cell morphology changes more pronounced. Wright Giemsa staining found that SAHA 64μmol·L-1 and OC3 / P cells incubated 24 h, the cell volume decreased, cytoplasm aggregation and nuclear pyknosis. MTT assay showed that SAHA 4 ~ 64μmo · lL-1 had an inhibitory effect on the survival of OC3 / P cells, and there was a significant time-concentration effect. SAHA 64μmol·L-1 for 6,12,24 and 48 h The inhibitory rates of OC3 / P cell survival were (7.1 ± 1.9)%, (14.6 ± 2.0)%, (36.8 ± 2.7)% and (83.3 ± 3.0)% (28.8 ± 1.2)%, (34.8 ± 4.3)%, (52.3 ± 2.8)%, (61.3 ± 8)% respectively at 8, 16, 32 and 64 μmol·L- 4.9%) and (83.3 ± 3.0)% (r = 0.966, P <0.05). The results of flow cytometry showed that apoptosis of OC3 / P cells was induced by SAHA 4, 16 and 64μmo · L-1 for 48 h (P <0.05, P <0.01). Transmission electron microscopy showed that SAHA 4, 16 and 64μmo · L-1 for 24 h, OC3 / P nucleus typical apoptotic morphological changes. Conclusion SAHA can inhibit the survival and induce apoptosis of OC3 / P cells in paclitaxel resistant human ovarian cancer cells.