淫羊藿苷通过诱导细胞有丝分裂灾变抑制外阴鳞癌SW962细胞增殖

Icariin inhibits proliferation of vulvar squamous cell carcinoma SW962 cells by inducing mitotic cataclysm

目的:探讨淫羊藿苷(ICA)能否抑制外阴鳞癌SW962细胞增殖及其潜在机制。方法:选取外阴鳞癌SW962细胞系为研究对象,ICA处理后,通过相差显微镜及HE染色、DAPI染色方法观察细胞形态变化;MTT法检测细胞增殖;免疫荧光法检测α-微管蛋白(α-tubulin)和波形蛋白(Vimentin)表达及分布;流式细胞术检测细胞周期;Western blot检测有丝分裂灾变及凋亡相关蛋白表达。结果:ICA诱导SW962细胞核发生多核分裂,抑制细胞活性,并将细胞阻滞在G 2/M期。免疫荧光检测结果显示α-tubulin和Vimentin蛋白呈多极分布并凝聚在细胞核周围。ICA在12 h内显著上调有丝分裂相关蛋白Chk1、CDK1、cyclinB1和cdc25c表达,但在24~48 h持续下降。凋亡相关蛋白cleaved caspase-3表达持续升高。结论:ICA可诱导外阴鳞癌SW962细胞发生有丝分裂灾变,抑制细胞增殖,最终导致细胞凋亡。

Objective:To investigate whether icariin(ICA)can inhibit the proliferation of vulvar squamous cell carcinoma SW962 cells and its potential mechanism.Methods:The SW962 cell line of vulvar squamous cell carcinoma was selected as the research object.After ICA treatment,the morphological changes of cells were observed by phase contrast microscope,HE staining and DAPI staining;MTT assay was used to detect cell proliferation and the expression and distribution ofα-tubulin and Vimentin proteins were detected by immunofluorescence;the cell cycle was detected by flow cytometry.Western blot was used to detect the expression of proteins related to mitotic catastrophe and apoptosis.Results:ICA induced multinucleation of SW962 nucleus and inhibited cell activity,as well as blocked cells cycle on G2/M phase.The results of immunofluorescence test thatα-tubulin and Vimentin proteins were multipolar and condensed around the nucleus.ICA significantly increased the expressions of mitosis related proteins Chk1,CDK1,cyclinB1 and cdc25c within 12 h,but continued to decline during 24~48 h.Expression of apoptosis related protein cleaved caspase-3 was increased continuously.Conclusion:ICA induces mitotic catastrophe,inhibits cell proliferation and eventually leads to cell apoptosis in SW962 cells.