天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (3): 294-297.doi: 10.11958/20150144

• 细胞与分子生物学 • 上一篇    下一篇

氯化钴低氧对黑色素瘤细胞系迁移能力及FSTL1表达分泌的影响

任方元1,李莲1,姜芳馨2,冯靖3,陈宝元1,曹洁3   

  1. 1 天津医科大学总医院呼吸科（邮编 300052）；2南开大学生命科学院
  • 收稿日期:2015-09-01 修回日期:2015-11-12 出版日期:2016-03-15 发布日期:2016-03-15
  • 通讯作者: 曹洁 E-mail:renfangyuan1020@126.com
  • 作者简介:任方元（1990）， 女， 硕士在读， 主要从事睡眠低氧性疾病和肿瘤相关研究
  • 基金资助:
    国家十二五科技支撑计划

Effect of cobalt chloride-induced hypoxia on cell migration, expression and secretion of FSTL1 in melanoma cell lines

REN Fangyuan， LI Lian， JIANG Fangxin， FENG Jing， CHEN Baoyuan , CAO Jie   

  1. 1 Respiratory Department of Tianjin Medical University General Hospital， Tianjin 300052, China; 2 College of Life Sciences, Nankai University
  • Received:2015-09-01 Revised:2015-11-12 Published:2016-03-15 Online:2016-03-15
  • Contact: CAO Jie E-mail:renfangyuan1020@126.com

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摘要： 目的 探讨氯化钴（CoCl2）化学模拟低氧对黑色素瘤细胞系B16F10迁移的影响，以及Follistatin-like 1（FSTL1）蛋白在此过程中的转录、表达和分泌情况。方法 CoCl2模拟低氧作用于小鼠B16F10细胞，实验分为3组：0 μmol/L CoCl2对照组、50 μmol/L和100 μmol/L CoCl2处理组。用MTT法测定细胞活力；用Transwell法测定细胞迁移能力；qRT-PCR检测Fstl1 mRNA表达；Western blot检测细胞内外FSTL1蛋白表达。结果 CoCl2模拟低氧可以导致B16F10细胞存活率显著下降，并呈浓度和时间依赖性；50 μmol/L CoCl2处理组（0.158±0.006）、100 μmol/LCoCl2处理组（0.203±0.002）B16F10细胞迁移能力均明显高于对照组（0.107±0.001，均P<0.05）；50 μmol/L CoCl2处理组（1.573±0.114）、100 μmol /L CoCl2处理组（2.219±0.085）Fstl1 mRNA表达均显著高于对照组（0.962±0.054，均P<0.05），而胞内FSTL1蛋白表达与Fstl1 mRNA表达趋势一致。同时也发现，CoCl2处理组细胞外FSTL1蛋白表达均低于对照组，且100 μmol/L CoCl2处理组几乎检测不到FSTL1表达。结论 CoCl2模拟低氧促进黑色素瘤细胞迁移，可能与FSTL1的表达和分泌有关，但其功能和作用机制还需进一步探讨。

关键词: 黑色素瘤, 细胞运动, 细胞低氧, 氯化钴, 迁移, FSTL1

Abstract: Objective The aim of this study was to explore the effect of cobalt chloride-induced hypoxia on migration of melanoma cells, and detect the transcription, expression and secretion of FSTL1 in this process. Methods B16F10 melanoma cell lines were treated with CoCl2 in order to mimic hypoxia. Experiment was divided into three groups: 0 μmol/L,50 μmol/L ,100 μmol/L .MTT assay was used to assure cell viability and determine the treatment concentration of CoCl2. Transwell assay was used to determine the migration ability of B16F10 melanoma cell line. Real-time PCR was used to measure the mRNA expression of FSTL1. Western blot were used to detect the protein expression of intracellular and extracellular of FSTL1. Results The cell viability of B16F10 melanoma cell lines was significantly reduced by CoCl2 treatment, in a time and concentration-dependent manner; The migration ability of B16F10 cell lines in CoCl2 treated group was significantly increased compared with the control group（all P<0.05）; The mRNA level of Fstl1 in CoCl2 treated group were obviously higher than the control group （all P<0.05）, the same as the protein of FSTL1. Simultaneously, the extracellular protein level of FSTL1 was significantly decreased compared with the control group and even it was not found in 100umol/L CoCl2 treatment. Conclusion The migration ability of melanoma cell lines was enhanced by CoCl2 treatment, which may be associated with expression and secretion of FSTL1, however, the relevant mechanism still be explored in the future.

Key words: melanoma, cell movement, cell hypoxia, cobalt chloride, migration, follistatin-like 1