天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (9): 970-974.doi: 10.11958/j.issn.0253-9896.2015.09.003

• 细胞与分子生物学 • 上一篇    下一篇

MCL-1和FBW7蛋白在纺锤丝毒性药物诱导的乳腺癌多倍体中的表达及临床意义

张倩1,袁碧波1,王彦1,许毅2   

  1. 1. 天津医科大学总医院
    2. 河北省清华发展研究院
  • 收稿日期:2015-03-10 修回日期:2015-05-08 出版日期:2015-09-15 发布日期:2015-09-15
  • 通讯作者: 张倩 E-mail:zhangqian19880303@163.com
  • 作者简介:张倩(1988)， 女， 硕士在读， 主要从事妇科肿瘤耐药机制的研究
  • 基金资助:
    国家自然科学基金资助项目(30872969)

Expression and clinical significance of MCL-1 and FBW7 proteins in breast cancer polyploid induced by spindle poisons

ZHANGQian1,YUANBibo1 ,WANGYan1,XUYi2   

  1. 1 Department of Gynecology and Obstetrics, Tianjin Medical University General Hospital,Tianjin 300052,China; 2 Hebei Tsinghua Development Research Institute
  • Received:2015-03-10 Revised:2015-05-08 Published:2015-09-15 Online:2015-09-15

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摘要： 摘要:目的 探讨 MCL-1 和 FBW7 蛋白在纺锤丝毒性药物诱导的乳腺癌多倍体中的表达及临床意义。方法（1） 以纺锤丝毒性药物 Nocodazole 处理人乳腺癌 MDA-MB-231 细胞， 显微镜下观察细胞形态学变化， 并于 0、 6、 12、 24、 48 及 72 h 收获细胞， 流式细胞术检测细胞周期和染色体倍体变化， Western blot 检测细胞中 FBW7 和 MCL-1 蛋白的表达。（2） 用多激酶抑制剂索拉非尼 （Sorafenib） 分别与 Nocodazole、 紫杉醇 （Taxol） 联合或单独处理细胞，Western blot 检测处理 48 h 后各组细胞中 MCL-1 蛋白的表达， 流式细胞术检测处理 48 h 细胞周期和染色体倍体变化，MTT 法检测处理 48、 72 h 细胞增殖情况。结果 （1） Nocodazole 处理后， 细胞出现体积增大、 核大的多倍体形态学改变， 0、 6、 12、 24、 48 及 72 h 八倍体细胞所占百分比分别为 （0.8±0.2） %、（8.5±2.3） %、 （7.8±2.0） %、 （9.9±0.9） %、 （28.2± 0.8） %及 （35.1±4.9） %， 逐渐增加 （P < 0.001）， 48 h 后多倍体 （四倍体和八倍体） 细胞数高达 （97.6±0.7） %； 随时间延长， FBW7 蛋白表达量减少， MCL-1 蛋白表达量增加。（2） 处理细胞 48 h 后， Nocodazole+Sorafenib 组较 Nocodazole 组、 Taxol+Sorafenib 组较 Taxol 组 MCL-1 蛋白表达量均明显减少， 多倍体细胞均减少， 细胞生长增殖率下降 （P < 0.05）。结论 FBW7 蛋白低表达， MCL-1 蛋白高表达与乳腺癌多倍体细胞的形成密切相关； Sorafenib 抑制 MCL-1 蛋白表达， 可能减少多倍体肿瘤形成。

关键词: 乳腺肿瘤, 多倍性, 体外研究, MCL-1, FBW7, 纺锤丝毒性药物, Nocodazole, 索拉非尼

Abstract: Abstract: Objective To investigate the expression and clinical significance of myeloid cell leukemia-1 (MCL-1) and F-box and WD repeat domain-containing 7 (FBW7) in breast cancer polyploid induced by spindle poisons. Methods (1) Nocodazole spindle poison was used to treat breast cancer cell MDA-MB-231. The morphological changes of cells were ob⁃ served under microscope, and cells were harvested in 0, 6, 12, 24, 48 and 72 h. The cell cycle and DNA-ploidy changes were examined by flow cytometry. The expressions of FBW7 and MCL-1 proteins were detected by Western blot assay. (2) A multikinase inhibitor (Sorafenib) with Nocodazole or Taxol was used to treat MDA-MB-231 cells. MCL-1 protein expression was detected by Western blot assay after 48 h treatment. The cell cycle and DNA-ploidy changes were examined by flow cy⁃ tometry after 48 h treatment. MTT method was used to observe cell proliferation after 48 and 72 h treatment. Results (1)Af⁃ ter treatment by Nocodazole, polyploid characteristics of large cell size and nucleus were appeared. The percentages of octa⁃ ploid were (0.8±0.2)%, (8.5±2.3)%, (7.8±2.0)%, (9.9±0.9)%, (28.2±0.8)% and (35.1±4.9)% after 0, 6, 12, 24, 48 and 72 h treatment, showing the increasing trend in turn (P < 0.001). The number of polyploidy (tetraploid and octaploid) cells was as high as (97.6±0.7)% after 48 h treatment. The expression level of FBW7 protein was decreased significantly but the expres⁃ sion of MCL-1 protein was increased significantly after 48 h treatment. (2) After 48 h treatment, the expression level of MCL- 1 protein, polyploidy percentage and cell proliferation decreased significantly in Nocodazole+Sorafenib group and Taxol+Sorafenib group compared with those of Nocodazole group and Taxol group (P < 0.05). Conclusion The lower expression of FBW7 protein and over- expression of MCL- 1 protein are correlated with the formation of breast cancer polyploidy. Sorafenib can reduce polyploid tumor cells by inhibiting MCL-1 protein expression.

Key words: breast neoplasms, polyploidy, in vitro, myeloid cell leukemia-1, F-box and WD repeat domain-containing 7, spindle poison, Nocodazole, sorafeni