Leukamenin E诱导人宫颈癌HeLa细胞胞质骨架重排和迁移抑制的研究  

Leukamenin E-induced cytoskeletal rearrangement and migration inhibition in HeLa cells

作　　者：杜世龙 宁秀梅 何苗[1] 李翡翡[1] 张会[1] 丁兰[1] DU Shi-long;NING Xiu-mei;HE Miao;LI Fei-fei;ZHANG Hui;DING Lan(College of Life Sciences,Northwest Normal University,Lanzhou 730070,China)

机构地区：[1]西北师范大学生命科学学院,兰州730070

出　　处：《天然产物研究与开发》2021年第7期1186-1197,共12页Natural Product Research and Development

基　　金：国家自然科学基金(31660101,30960464)。

摘　　要：为了探索对映-贝壳杉烷二萜leukamenin E对细胞骨架和细胞迁移的影响,该文采用MTT法、吉姆萨染色、流式细胞术及免疫荧光技术研究了leukamenin E诱导HeLa细胞的3种胞质骨架重排,细胞迁移抑制及可能的作用机制。结果显示,0.4~1.0μmol/L的leukamenin E可导致G1期阻滞,抑制细胞增殖,使细胞形态及核形态发生显著变化,细胞伪足减少以及“肾形核”细胞数量增多,并抑制细胞迁移;Leukamenin E可显著改变HeLa细胞微管和角蛋白纤维的排布,诱导微管和角蛋白纤维在核周聚集,但减少胞质中应力纤维数量。Leukamenin E显著升高HeLa细胞内活性氧(ROS)水平,但加入活性氧清除剂NAC可明显减弱leukamenin E诱导的细胞形态改变、细胞骨架重排以及细胞迁移的抑制作用,表明活性氧途径与上述变化相关联。上述结果说明,leukamenin E通过ROS水平升高调节相关信号途径,导致HeLa细胞骨架重排而引起细胞及细胞核形态改变,同时导致了细胞的迁移的抑制效应。In order to explore the effects of leukopenia E,an ent-kaurane diterpenoid on cytoskeleton and cell migration,leukamenin E-induced cytoskeletal rearrangement and inhibition of cell migration were investigated by using MTT,Giemsa staining,flow cytometry and immunofluorescence assay.The results showed that 0.4-1.0μmol/L leukamenin E inhibited the proliferation of HeLa cells through G1 arrest,led to obvious change in cellular morphology and nuclear morphology,while reduced the number of pseudopods and increased the number of flattened nucleus.Moreover,the migration of HeLa cells was inhibited.Leukamenin E could significantly alter the arrangement of microtubules and keratin filaments,decrease stress filaments in the cytoplasm,and the aggregation of microtubules and keratin filaments into large bundles around the nucleus have been showed.Leukamenin E induced a significant increase in intracellular ROS levels.A radical scavenger NAC,reduced leukamenin E-induced cell morphology changes,cytoskeletal rearrangement and inhibition of cell migration,namely,these changes was associated with the increase of ROS.These results suggested that upregulation of ROS level induced by leukameninE resulted in the rearrangement of HeLa cytoskeleton by regulating the related signal pathway,which leads to morphological changes of cells and nuclear,and inhibits cell migration.

关 键 词：对映贝壳杉烷二萜 Leukamenin E 细胞质骨架 细胞迁移 活性氧 

分 类 号：R285.5[医药卫生—中药学]