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吗啡联合顺铂对卵巢癌细胞Skov3增殖、凋亡及侵袭转移的影响

阮志平1,郑华2,孙志华3   

  1. 1 100039 北京 北京市朝阳区妇幼保健院麻醉科 2 100039 北京市朝阳区妇幼保健院妇科 3 100039 北京市朝阳区妇幼保健院妇瘤科
  • 收稿日期:2015-10-28 修回日期:2015-11-02 出版日期:2016-01-31 发布日期:2016-01-31

Combined effect of morphine and cisplatin on proliferation, apoptosis, as well as invasion of ovarian cancer cell line Skov3

RUAN Zhiping, ZHENG Hua, SUN Zhihua   

  1. Department of Anesthesia, Maternal and Child Health Care of Chaoyang District, Beijing 100039, China
  • Received:2015-10-28 Revised:2015-11-02 Online:2016-01-31 Published:2016-01-31

摘要: 目的 探讨吗啡(Mor)联合顺铂(DDP)对人卵巢癌Skov3细胞增殖、凋亡及侵袭转移的影响。方法 采用MTT法观察不同浓度Mor(10、20、50、100、200 μmol/L)和DDP(1、2、5、10、20 mg/L)作用于Skov3细胞24、48、72、96 h后的增殖抑制率,筛选适合的浓度进行后续功能学研究。取对数生长期Skov3细胞分为4组:Mor(200 μmol/L)组、DDP(20 mg/L)组、Mor+DDP(200 μmol/L Mor+20 mg/L DDP)组和未加药对照组,分别采用流式细胞仪和实时定量PCR检测各组处理48 h的细胞凋亡率及凋亡相关基因(Bax、Bcl-2和caspase-3)的mRNA水平,采用Transwell试验和Western blotting检测各组处理48 h后穿膜细胞数和侵袭相关蛋白(MMP-2、MMP-9、TIMP-1和TIMP-2)水平。结果 随Mor(10~200 μmol/L)和DDP(1~20 mg/L)浓度增加,Skov3细胞的增殖能力受抑制,且抑制效应呈浓度和时间依赖方式;Mor+DDP组处理24、48、72、96 h后的增殖抑制率高于Mor组、DDP组和对照组(P<0.05)。与对照组相比,Mor、DDP和Mor+DDP组的细胞凋亡率、caspase-3活化率、Bax mRNA、caspase-3 mRNA、TIMP-1和TIMP-2水平均升高,Bcl-2 mRNA、穿膜细胞数、MMP-2和MMP-9水平均降低,差异有统计学意义(P<0.05);Mor+DDP组的上述指标水平均优于Mor组和DDP组(P<0.05)。结论 Mor和DDP对Skov3细胞均有细胞毒性,能抑制增殖及侵袭转移并诱导凋亡,且Mor联合DDP应用的抑制作用增强。

Abstract: ObjectiveTo investigate the effect of morphine(Mor)combined with cisplatin(DDP)on the proliferation,apoptosis, as well as invasion and metastasis of human ovarian cancer Skov3 cells. Methods MTT method was used to observe the cell survival rates at 24,48,72,96 h after the treatment of mor at different concentrations(10,20,50,100,200 μmol/L)and DDP(1,2,5,10 and 20 mg/L) as to screen the concentration of Mor and DDP for subsequent functional study. Skov3 cells at logarithmic growth phase were divided into 4 groups:Mor(200 μmol/L)group,DDP(20 mg/L)group,Mor(200 μmol/L)+DDP(20 mg/L) group and control group(Skov3 cells without treatment). Flow cytometry and real-time quantitative PCR were used to detect the apoptosis rate and mRNA levels of apoptosis related genes(Bcl-2,caspase-3 and Bax)at 48 h after the treatment. Transwell test and Western blotting were used to detect the number of cells and the invasion associated proteins(TIMP-2,TIMP-1 MMP-9 and MMP-2)at 48 h after treatment. Results The proliferation inhibition rates increased gradually with the increase of Mor(10-200 μmol/L)and DDP(1-20 mg/L)concentration. The inhibitory effect was in a time and concentration dependent manner. The proliferation inhibition rates of Mor+DDP group was higher than those of Mor group,DDP group and control group at 24, 48,72,96 h after treatment(P<0.05). Compared with control group,there were increased apoptosis rate,caspase-3 activation rate and mRNA levels of Bax and caspase-3, as well as protein levels of TIMP-1 and TIMP-2,but decreased Bcl-2 mRNA,number of cells penetrating membrane and protein levels of MMP-9 and MMP-2 in cells treated with Mor or DDP alone or in combination(P<0.05). The levels of the above indexes in Mor+DDP group were better than those in Mor or DDP groups(P<0.05). Conclusion Both Mor and DDP can inhibit the proliferation of Skov3 cells and induce apoptosis,and the inhibition of Mor combined with DDP is enhanced.

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