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山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (2): 27-32.

• 基础医学 • 上一篇    下一篇

PD98059联合顺铂对卵巢癌细胞增殖的影响

贾宗洋,张向宁,靳艳慧,魏美玲   

  1. 山东大学齐鲁医院妇产科,  济南 250012
  • 收稿日期:2012-11-15 出版日期:2013-02-10 发布日期:2013-02-10
  • 通讯作者: 张向宁(1960- ),男,主任医师,硕士生导师,主要从事妇科肿瘤和妇科微创的研究。E-mail:xningzhang@yahoo.com.cn
  • 作者简介:贾宗洋(1987- ),男,硕士研究生,主要从事妇科肿瘤的研究。E-mail:fcjzy87@163.com

Effects of PD98059 combined with cisplatin derivative on human ovarian carcinoma cell SKOV3

JIA Zong-yang, ZHANG Xiang-ning, JIN Yan-hui, WEI Mei-ling   

  1. Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan 250012, China
  • Received:2012-11-15 Online:2013-02-10 Published:2013-02-10

摘要:

目的   观察MEK1/2特异性抑制剂PD98059联合顺铂对卵巢癌SKOV3细胞增殖的影响。 方法   按不同处理因素对卵巢癌SKOV3细胞分组:空白对照组、单纯PD98059组、单纯顺铂组、联合用药组。CCK-8法检测各组细胞的增殖抑制率。实时定量PCR检测各组细胞含有WW结构域的氧化还原酶(WWOX)和细胞外调节蛋白激酶(ERK)在基因水平的表达。Western-blot检测各组细胞磷酸化ERK1/2(p-ERK1/2)蛋白的表达。光镜下观察各组细胞的生长状态及形态学改变。流式细胞术检测各组细胞的凋亡情况。结果   联合用药组细胞增殖抑制率明显高于其他各组(P<0.05);单纯顺铂组及联合用药组WWOX mRNA表达量较空白对照组明显增加(P<0.05),单纯PD98059组WWOX表达量无明显改变(P>0.05);ERK mRNA在各组间的表达无明显变化(P>0.05);与空白对照组相比,p-ERK1/2表达量在单纯PD98059组及联合用药组中明显降低(P<0.05),在单纯顺铂组的表达无明显变化。光镜下可观察到空白对照组及单纯PD98059组细胞生长状态好,细胞密集,无明显形态学改变;单纯顺铂组及联合用药组贴壁细胞少,细胞稀疏,联合用药组部分细胞出现胞浆溶解样改变。结论   PD98059联合顺铂可能通过调控WWOX基因表达及ERK通路,对卵巢癌细胞增殖产生较强的抑制作用。

关键词: 卵巢肿瘤;顺铂;WW结构域;氧化还原酶;细胞外调节蛋白激酶

Abstract:

Objective   To investigate the effects of MEK/ERK inhibitor PD98059, cisplatin and the combined treatment on the proliferation of human ovarian carcinoma cell SKOV3, and to investigate the potential mechanism. Methods   Cell Counting Kit-8(CCK-8)was used to investigate the SKOV3 cell proliferative inhibition rates of different groups: the blank control group, the cisplatin group, the PD98059 group and the combined treatment group. Real-Time PCR was used to detect the expression levels of WW domain-containing Oxidoreductase (WWOX) and extracellular regulated protein kinases (ERK) mRNA. Western blotting analysis was used to measure the expression of phosphorylated ERK1/2(p-ERK1/2) in different groups. The morphological changes of various groups were observed under the light microscope. Flow cytometry was employed to analyze the apoptosis rate of SKOV3 cells in different groups. Results   The inhibitory effect on the proliferation of SKOV3 cells in the combined treatment group was more significant compared with the other three groups (P<0.05). RT-PCR analysis showed that the expressions of WWOX mRNA were increased in the cisplatin group and the combined treatment group compared with the other two groups. The WB analysis showed that the expressions of p-ERK1/2 in the PD98059 group and the combined treatment group were higher compared with the other two groups. More adherent cells were observed in the blank control group and the PD98059 group under the light microscope. There were less adherent cells in the cisplatin group and the combined treatment group, and cytoplasmic dissolution of cells could be observed under the light microscope in these two groups. Conclusion   PD98059 combined with cisplatin exhibited significant inhibitory effect on the proliferation of SKOV3 cells via regulating the expression of WWOX gene and the ERK pathway.

Key words: Ovarian neoplasms; Cisplatin; WW domain-containing; Oxidoreductase; Extracellular Regulated protein Kinases

中图分类号: 

  • R737.31
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