摘要
目的观察5-烯丙基-7-二氟亚甲基白杨素(ADFMChR)诱导人卵巢癌(CoC1)细胞凋亡的作用。方法以体外培养的人卵巢癌CoC1为研究对象。采用软琼脂克隆测定ADFMChR对细胞集落的影响;流式细胞术(FCM)检测ADFMChR诱导细胞凋亡率;凝胶电泳观察ADFMChR诱导基因DNA梯形条带。Western blot分析ADFMChR对CoC1细胞PPAR,γNF-KB,Bcl-2,Bax蛋白表达的影响。结果软琼脂克隆显示ADFMChR呈剂量依赖性抑制细胞集落形成;FCM分析发现ADFMChR呈剂量依赖性诱导细胞凋亡;ADFMChR(30μmol/L)孵育CoC1细胞48h后,DNA琼脂糖凝胶电泳呈现典型梯形条带。Western blot分析结果表明ADFMChR以剂量依赖方式上调CoC1细胞PPARγ和Bax蛋白表达,下调NF-κB和Bcl-2蛋白表达。结论ADFMChR诱导人卵巢癌CoC1细胞凋亡与其活化PPARγ,抑制NF-KB表达和提高Bax/Bcl-2比值有关。
Objective To investigate induction of apoptosis of human ovarian cancer CoC1 cells by 5-Allyl-7-Gen- Difluoromethylenechrysin （ADFMChR） in vitro,and its molecular mechanism. Methods The growth inhibition of CoCl cells by treatment with ADFMChR was used to evaluate by clone-formation in soft agar assay. The apoptosis of CoCl cells induced by ADFMChR was determined by DNA fragment assay and flow cytometry with PI staining. Effect of ADFMChR on expression of PPART,NF-κB,Bcl-2 and Bax protein of CoCl cells was detected by western blotting. Results The clone formation of CoCl cells could be significantly inhibited by ADFMChR in a dose-dependent manner. ADFMChR significantly induced apoptosis of CoCl cells in a concentration- dependent manner. DNA fragment assay indicated that DNA ladder bands could be shown in DNA agarose gel electrophoresis after treatment with ADFMChR at 30.0μmol/L for 48h. Western blot analysis shown that expression of PPAR7 and Bax proteins were upregulation and protein of NF-κB and Bcl-2 were downregulation after treatment with ADFMChR in a＇ concentration-dependent manner. Condusion The effect of ADFMChR on induction of apoptosis of CoCl cells may be associated with activation of PPARγ , sequentially accompanied by inhibition of protein expression of NF-κB and increase ratio of Bax to Bcl- 2.
出处
《中国比较医学杂志》
CAS
2008年第3期5-9,共5页
Chinese Journal of Comparative Medicine
基金
湖南省自然科学基金资助项目(03JJY5009)
