内质网功能状态对卵巢癌细胞顺铂敏感度的影响

发布时间：2018-07-13 15:27

【摘要】：目的探讨内质网应激(endoplasmic reticulum stress,ERS)功能状态对卵巢癌细胞顺铂(DDP)敏感度的影响。方法卵巢癌细胞系SKOV3接受40μmol/L DDP处理24 h后,q RT-PCR检测ERS相关基因改变;同时内质网示踪染料(ER-tracker)观察内质网形态变化;分别用ERS稳定剂牛磺熊去氧胆酸钠(tauroursodeoxycholate,TUDCA)或ERS诱导剂毒胡萝卜素(thapsigargin,TG)预处理SKOV3,再接受DDP处理,应用流式细胞术检测细胞凋亡率;应用Western blot法检测内质网应激相关蛋白GRP78和CHOP是否表达;从临床卵巢癌患者腹水中分离与培养原代卵巢癌细胞后,采用流式细胞术比较TUDCA及TG预处理对原代卵巢癌细胞DDP敏感度影响,Western blot法检测凋亡相关蛋白PARP和Caspase-3的变化。结果顺铂处理卵巢癌细胞系SKOV3后能够引起明显的ERS,ERS相关蛋白明显上调,内质网形态呈现颗粒化;TUDCA可以显著下调DDP引起的杀伤作用(P0.05),显著降低GRP78和CHOP蛋白水平;TG可以显著上调DDP引起的杀伤作用(P0.05)、GRP78和CHOP蛋白水平;在卵巢癌腹水原代细胞中TUDCA亦可显著抑制DDP引起的杀伤作用(P0.05),TG亦可显著上调DDP引起的杀伤作用(P0.05)。结论改变ERS的功能状态能够影响卵巢癌细胞对顺铂的敏感度。
[Abstract]:Objective to investigate the effect of endoplasmic reticulum stress (ERS) on the sensitivity of cisplatin (DDP) in ovarian cancer cells. Method ovarian cancer cell line SKOV3 was treated with 40 u mol/L DDP for 24 h, Q RT-PCR detected the changes of ERS related genes; and endoplasmic reticulum tracer dye was used to observe the morphologic changes of endoplasmic reticulum; The stabilizer tauroursodeoxycholate, TUDCA, or ERS inducer, thapsigargin, TG, pretreated SKOV3, and then DDP treatment, and the cell apoptosis rate was detected by flow cytometry; Western blot method was used to detect the expression of egg white GRP78 and CHOP in endoplasmic reticulum stress; from the ascites of the patients with ovarian cancer. After isolation and culture of primary ovarian cancer cells, flow cytometry was used to compare the effect of TUDCA and TG preconditioning on the DDP sensitivity of primary ovarian cancer cells. Western blot assay was used to detect the changes of apoptosis related protein PARP and Caspase-3. Results cisplatin treated ovarian cancer cell line SKOV3 could cause obvious ERS, ERS related protein obviously up, endoplasmic reticulum TUDCA could significantly reduce the killing effect (P0.05) caused by DDP and significantly decrease the level of GRP78 and CHOP protein; TG can significantly increase the killing effect (P0.05), GRP78 and CHOP protein levels caused by DDP, and also significantly inhibit the killing effect caused by DDP in the primary cells of ovarian cancer ascites. The killing effect of DDP (P0.05). Conclusion changing the functional state of ERS can affect the sensitivity of ovarian cancer cells to cisplatin.
【作者单位】：华中科技大学同济医学院附属同济医院肿瘤生物医学中心;
【基金】：国家自然科学基金面上项目(30973184)
【分类号】：R737.31

【共引文献】