【摘 要】
：
目的 通过体外实验探讨铂耐药的上皮性卵巢癌细胞中是否存在Notch信号通路异常激活及EMT样表型的获得,并探讨γ-分泌酶抑制剂对化疗耐药EOC增殖、粘附、侵袭、转移等生物学行为的影响,为卵巢癌治疗提供新思路.方法 体外培养人卵巢浆液性囊腺癌细胞株SKOV3及其顺铂耐药株SKOV3/DDP,PCR检测其Notch1 mRNA表达量的差异,western-blot检测γ分泌酶抑制剂DAPT作用于SKO
【机 构】
：
广东省妇产医院·广东省妇幼保健院 广州511442
【出 处】
：
广东省遗传学会第九届代表大会暨学术研讨会
论文部分内容阅读
目的 通过体外实验探讨铂耐药的上皮性卵巢癌细胞中是否存在Notch信号通路异常激活及EMT样表型的获得,并探讨γ-分泌酶抑制剂对化疗耐药EOC增殖、粘附、侵袭、转移等生物学行为的影响,为卵巢癌治疗提供新思路.方法 体外培养人卵巢浆液性囊腺癌细胞株SKOV3及其顺铂耐药株SKOV3/DDP,PCR检测其Notch1 mRNA表达量的差异,western-blot检测γ分泌酶抑制剂DAPT作用于SKOV3/DDP细胞前后,E-cadherin及vimentin的表达情况,划痕实验及Transwell试验检测其迁移侵袭能力改变.结果 1倒置显微镜下观察,SKOV3细胞呈上皮样细胞形态；SKOV3/DDP细胞呈间质细胞样形态；Western-blot：与SKOV3相比,SKOV3/DDP细胞的E-cadherin表达量低,Vimentin表达量高(P＜0.01)；RT-PCR结果示SKOV3/DDP细胞Notch1 mRNA的表达量为SKOV3细胞的3.952倍.2 MTT：随着DAPT浓度增加和作用时间的延长,SKOV3/DPP细胞的生长抑制率明显升高(P＜0.01).3 Western-blot：以DAPT (25、50、75 μ mol/L)作用于SKOV3/DDP细胞48h后,E-cadherin表达较对照组明显增加(P＜0.01),各组间差异显著(P＜0.01)；vimentin表达较对照组明显降低(P＜0.01),但50、75μ mol/L浓度组差异无统计学意义(P＞0.05).4 50μ mol/L DAPT作用于SKOV3/DDP细胞24h,划痕试验结果：处理组的划痕愈合面积显著小于对照组(P＜0.01)；Transwell迁移及侵袭试验结果示处理组穿膜细胞个数明显少于对照组(P＜0.01).结论 SKOV3/DDP细胞化疗耐药过程中获得了EMT样表型,γ-分泌酶抑制剂DAPT可通过靶向作用于Notch信号通路,部分逆转EMT,抑制人卵巢癌顺铂耐药细胞的增殖、粘附及侵袭转移能力.
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