摘要
目的:观察顺铂对子宫内膜癌Ishikawa细胞的增殖及自噬的影响。方法:MTS法检测不同剂量顺铂对Ishikawa细胞增殖的作用。透射电镜观察自噬泡形成,应用Western blot检测LC3及Beclin1的积累,免疫荧光检测LC3的荧光聚集情况。结果:(1)10μg/m L顺铂对Ishikawa细胞的增殖有抑制作用,随着时间及浓度的增加,细胞增殖的抑制作用明显增加(P<0.01);(2)透射电镜显示顺铂处理后子宫内膜癌细胞发生自噬,随着时间剂量增加,Western blot检测LC3表达增加,Becline1表达无明显变化;(3)与对照组相比,20μg/m L-12 h组的自噬体及LC3表达增加;20μg/m L-24 h组较12 h组的自噬体及LC3表达增加。结论:顺铂对Ishikawa细胞增殖有明显抑制作用,呈时间剂量依赖性;顺铂诱导Ishikawa细胞发生自噬,呈时间剂量依赖性。自噬相关基因LC3的变化可能为顺铂诱导子宫内膜癌自噬分子机制之一。
Objective To explore the effects of cisplatin on the proliferation and autophagy of endometrial carcinoma Ishikawa cells, Methods Ishikawa cell proliferation was detected by MTS assay after the cells were treated with CDDP. To assess the level of autophagy, transmission electron microscope and Western blot were used to detect LC3 and Beclinl expression; fluorescence microscopy was used to observe the fluorescence aggregation of green fluorescent protein and microtubule associated protein 1 light chain 3 fusionprotein （GFP-LC3）. Results Cisplatin of 10 g/mL inhibited the proliferation of Ishikawa cells, with an increase of time and concentration, the inhibition of cell proliferation was significantly elevated （P 〈 0.01）. Transmission electron microscopy showed that under a condition of cisplatin on Ishikawa endometrial cancer cell, autophagy occurred. With an increase of concentration and dosage, Western blot showed that autophagy related protein LC3 expression was up-regulated, but becline-1 had no obvious change. LC3 expression level was higher in 12h-treatment with 20 μg/mL cisplatin group than in the control group, and was higher in 24h-treatment group than in 12h-treatment group. Conclusions Cisplatin inhibits proliferation of Ishikawa ceils and induces autophagy of the ceils in a time-and dose-dependent manner. Autophagy related MAP-LC3 is involved in the molecular mechanisms of autophagy induced by cisplatin in endometrial carcinora.
作者
林琼燕
王沂峰
熊汉真
生秀杰
LIN Qiongyan WANG Yifeng XIONG Hanzhen SHENG Xiujie(Third Affiliated Hospital, Guangzhou Medical Universi- ty, Guangzhou 510515, China)
出处
《实用医学杂志》
CAS
北大核心
2017年第4期524-528,共5页
The Journal of Practical Medicine
基金
广州市卫生局一般引导项目资助项目(编号:2015A013010068)
关键词
顺铂
ISHIKAWA细胞
增殖
自噬
自噬相关基因
Cisplatin
Endometrial cancer cells
Proliferation
Autophagy
Autophagy related genes
