摘要
目的探讨多不饱和脂肪酸DHA对宫颈癌细胞(He La、Siha)凋亡、迁移侵袭的影响。方法 He La、Siha细胞,分为空白对照组、不同浓度DHA组,观察不同药物浓度作用于宫颈癌细胞后的形态学改变,Hoechst 33258染色检测48 h后细胞核凋亡情况;MTT法检测DHA作用于宫颈癌细胞24、48、72 h后的细胞凋亡情况;流式细胞仪检测不同浓度DHA作用于宫颈癌细胞48 h后的凋亡率;细胞划痕实验和Transwell迁移实验检测不同浓度DHA对宫颈癌细胞体外迁移能力的影响;Western Blotting检测DHA刺激细胞48 h后凋亡相关蛋白(Bax、Bcl-2、cleaved-caspase3)及肿瘤转移侵袭相关蛋白MMP-9、VEGF表达情况。结果倒置显微镜下及经Hoechst33258染色后,随着DHA浓度增加,宫颈癌细胞形态改变越明显,凋亡小体增多;MTT结果显示DHA呈时间-浓度依赖性抑制宫颈癌细胞增殖,促进其凋亡;细胞划痕实验和transwell迁移实验提示DHA作用于细胞后,浓度越高,对细胞的体外迁移能力的抑制作用越强;Western Blotting显示DHA作用于细胞48 h后,cleaved-caspase3、Bax蛋白表达增加(P<0.05),而Bcl-2、MMP-9、VEGF蛋白表达下降(P<0.05),Bcl-2/Bax比值下降(P<0.05)。结论 DHA可通过调控cleavedcaspase3、Bax及Bcl-2表达,促进宫颈癌细胞凋亡的作用并能通过降低MMP-9、VEGF的表达抑制宫颈癌细胞的迁移和侵袭。
Objective To investigate the effect of docosahexaenoic acid（DHA） on apoptosis, migration and invasion of cervical cancer cell lines. Methods cervical cancer cell lines Hela and Siha in logarithmic phase were treated different concentrations of DHA. The morphological changes of the cells were observed microscopically and cell apoptosis was observed using Hoechst33258 fluorescent staining. MTT assay was used to evaluate the effect of DHA in suppressing cell growth, and flow cytometry was employed to analyze the changes of cell apoptotic rate following DHA stimulations. Wound healing assay and Transwell migration assay were used to evaluate the migration of the cell lines. The expression levels of Bax, Bcl- 2 cleaved caspase3,MMP- 9 and VEGF proteins were detected by Western blotting. Results DHA exposure of the cells caused obvious morphological changes and dose- dependently increased the number of apoptotic bodies in the cells. MTT assay showed that DHA inhibited the growth of the cancer cells in a time- and concentration-dependent manner. DHA also effectively suppressed migration and invasion of the cancer cells. The cells exposed to DHA showed significantly down-regulation of Bcl-2, MMP-9and VEGF proteins and up- regulation of cleaved- caspase 3 and Bax. Conclusion DHA can promote cervical carcinoma cell apoptosis by down- regulating the anti- apoptotic proteins Bax, Bcl- 2 and cleaved- caspase3 and suppress cell invasion by decreasing MMP-9 and VEGF expressions.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2016年第6期848-856,共9页
Journal of Southern Medical University
基金
广东省科技计划项目(2016A020215115)
花都区科技计划产学研协同创新重点专项(HD15CXY006)
关键词
宫颈肿瘤
DHA
细胞凋亡
细胞侵袭
细胞转移
cervical carcinoma
docosahexaenoic acid
apoptosis
invasion
migration
