靶向敲除JARID2基因对葡萄膜黑色素瘤细胞生长与转移的影响
网络出版日期: 2015-06-04
基金资助
国家自然科学基金(81372469, 81172323)
Effects of targeted knockout of JARID2 gene on growth and migration of uveal melanoma cells
Online published: 2015-06-04
Supported by
National Natural Science Foundation of China, 81372469, 81172323
目的 探讨JARID2基因在葡萄膜黑色素瘤细胞生长和转移中的作用。方法 采用real-time PCR检测葡萄膜黑色素瘤细胞系OM431和SP6.5中JARID2 mRNA的表达。利用shRNA敲除OM431和SP6.5的JARID2基因,构建稳定转染细胞系shSP6.5和shOM431,Western blotting检测JARID2蛋白表达变化,观察靶向敲除JARID2基因对葡萄膜黑色素瘤细胞生长、转移及体外克隆形成的影响。结果 OM431和SP6.5细胞中JARID2 mRNA呈过表达。Western blotting检测结果显示:与OM431和SP6.5细胞比较,shOM431和shSP6.5细胞中JARID2蛋白表达显著减少。靶向敲除JARID2基因的葡萄膜黑色素瘤细胞的生长受到抑制,细胞转移数量减少,体外克隆形成率降低。结论 JARID2基因在葡萄膜黑色素瘤细胞的生长、转移和克隆形成能力中起着重要的作用。
刘博 , 邱纯 , 李鹏 , 等 . 靶向敲除JARID2基因对葡萄膜黑色素瘤细胞生长与转移的影响[J]. 上海交通大学学报(医学版), 2015 , 35(5) : 642 . DOI: 11.3969/j.issn.1674-8115.2015.05.004
Objective To investigate the role of JARID2 gene in the growth and migration of uveal melanoma cells. Methods The mRNA expression of JARID2 in uveal melanoma cell lines OM431 and SP6.5 was detected by the real-time PCR. The JARID2 gene of OM431 and SP6.5 was knocked out by shRNA and stable transfected cell lines shSP6.5 and shOM431 were constructed. Variations of protein expression of JARID2 were detected by the Western blotting. Effects of targeted knockout of JARID2 gene on the growth, migration, and in vitro colony formation of uveal melanoma cells were observed. Results The mRNA expression of JARID2 in OM431 and SP6.5 was over-expressed. The results of Western blotting showed that compared with OM431 and SP6.5, the protein expression of JARID2 in shOM431 and shSP6.5 significantly decreased. Targeted knockout of JARID2 gene could inhibit the growth, migration, and in vitro colony formation of uveal melanoma cells. Conclusion JARID2 gene plays an important role in the growth, migration, and colony formation of uveal melanoma cells.
Key words: JARID2 gene; uveal melanoma; tumor metastasis
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