摘要
目的观察土木香内酯对Hela细胞BMI1基因表达的影响,探讨AL抑制Hela细胞增殖的可能机制。方法体外培养人宫颈癌Hela细胞株,CCK8(cell counting kit⁃8)法和平板克隆实验检验AL对Hela细胞活性及增殖能力的影响;通过Real⁃time PCR与Western blot检测经AL处理过的Hela细胞BMI1基因的表达情况;Real⁃time PCR与western blot检测沉默和过表达BMI1基因的效率;过表达BMI1基因后检测AL对Hela细胞增殖的影响。结果AL能显著抑制Hela细胞的活性及增殖能力,且呈明显的剂量依赖性;Hela细胞经AL处理后,BMI1的转录和蛋白表达明显降低;过表达BMI1基因后,Hela细胞的活性增强,AL可抑制由BMI1过表达引起的细胞活性增强;shRNA干扰BMI1后,BMI1的表达显著降低,再施予AL,BMI1的表达降低更加明显。结论AL抑制宫颈癌细胞增殖,其机制可能是通过抑制BMI1的表达发挥作用。
Objective To observe the effects of Alantolactone(AL)on BMI1 gene expression in Hela cells,and to explore the possible mechanism of AL on inhibiting the proliferation of Hela cells.Methods Human cervical cancer Hela cell line was cultured in vitro,and the effect of AL on the activity and proliferation ability of Hela cells was tested by CCK8(cell counting kit⁃8)method and plate cloning formation experiment.The expression of BMI1 in AL⁃treated Hela cells was detected by Real⁃time PCR and Western blot.Then the alterations of BMI1 gene expression was detected by Real⁃time PCR and Western blot after silencing and overexpression.The effect of AL on the proliferation of Hela cells was detected by CCK8(cell counting kit⁃8)method after overexpression of BMI1 gene.Results AL significantly inhibited the activity and proliferation of Hela cells in a dose⁃dependent manner.Results from real⁃time PCR showed that the expression of BMI1 levels were significantly down⁃regulated in AL treated group.After the over⁃expression of BMI1 gene,the activity of Hela cells was enhanced,and AL inhibited the enhanced cell activity caused by the over⁃expression of BMI1.After shRNA interference with BMI1,the expres⁃sion of BMI1 was significantly reduced.The expression of BMI1 gene was significantly reduced in shRNA and AL co⁃treated group.Conclusion AL could effectively inhibit the proliferation of cervical cancer cells possibly by down⁃regulating BMI1 expression.
作者
谢丽霞
许倩倩
徐红霞
杜凯丽
桑明
孙晓东
XIE Lixia;XU Qianqian;XU Hongxia;DU Kaili;SANG Ming;SUN Xiaodong(Central Laboratory of Xiangyang First People′s Hospital,Hu⁃bei University of Medicine,Xiangyang 441000,China)
出处
《实用医学杂志》
CAS
北大核心
2020年第3期288-293,共6页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号：81903005)
湖北省自然科学基金面上项目(编号：2017CFB674,2018CFB701)
湖北省对外科技合作项目(编号：2019AHB068)
湖北省卫健委科研项目(编号：ZY2019F028)
襄阳市科技局科技项目(编号：2017033)
武当特色中药研究湖北省重点实验室(湖北医药学院)开放课题(编号：WDCM2018009)
湖北医药学院药护学院团队项目(编号：2017YHKT02)
湖北医药学院人才启动金项目(编号：2015QDJZR03)
