摘要
目的 研究呋喃二烯酮对人结直肠癌RKO细胞凋亡的影响。方法 将不同浓度的呋喃二烯酮溶液(50、100、200、300、400μmol·L^(-1))加入体外培养的RKO细胞中,处理24、48、72 h后,通过CCK-8法检测RKO细胞的活性;用DAPI染色和Annexin V-FITC/PI流式细胞术检测药物处理过后细胞凋亡情况;Western blotting检测RKO细胞Bcl-2、Bcl-xl、Bax、凋亡诱导因子(AIF)、细胞色素C、第二个线粒体衍生的半胱氨酸蛋白酶激活剂(Smac/DIABLO)及存活素的蛋白表达情况。结果 呋喃二烯酮处理后24、48、72 h呋喃二烯酮对RKO细胞的半数抑制浓度(IC50)分别为(248.13±5.79)、(154.26±4.87)和(117.40±5.14)μmol·L^(-1),对RKO细胞的毒性呈现时间依赖性。流式细胞术检测显示细胞凋亡率随药物浓度的增加而明显增加。不同浓度呋喃二烯酮作用于RKO细胞后,细胞活性明显受到抑制,且随着呋喃二烯酮浓度的上升,细胞活性显著降低(P<0.05)。呋喃二烯酮能够显著降低线粒体凋亡通路的相关蛋白Bcl-2、Bcl-xl和存活素的表达,升高Bax、AIF、Smac/DIABLO和细胞色素C的含量。结论 呋喃二烯酮能抑制RKO细胞增殖、诱导细胞凋亡,激活细胞内线粒体凋亡途径通路可能是其作用机制之一。
AIM To investigate the effects of furanodienone on the apoptosis of human colorectal cancer RKO cells. METHODS RKO cells were treated by furanodienone at the varying concentrations（50, 100, 200, 300, 400 μmol·L^（-1）） for 24, 48 and 72 h in vitro. The cell viabilities were detected by CCK8. DAPI fluorescent staining was used to observe the influences of furanodienone on the change of the nucleus. Annexin V-FITC/PI staining was used to investigate the apoptosis in RKO cells by flow cytometry. The protein expressions of Bcl-2,Bcl-xl, Bax, apoptosis inducing factor（AIF）, cytochorome C, Smac/DIABLO and survivin were detected by Western blotting. RESULTS The IC_（50） of furanodienone were（248.13 ± 5.79）,（154.26 ± 4.87） and（117.40 ±5.14） μmol·L^（-1） in RKO cells at 24, 48 and 72 h, showing time-depended toxicity of RKO cells. Flow cytometric analysis showed that the apoptotic rates of RKO were enhanced significantly with the increased concentrations of furanodienone. After the RKO cells were treated by furanodienone at the varying concentrations, cell activity was significantly inhibited. With the increased concentrations of furanodieone, cell activity decreased significantly（P〈0.05）. The related proteins including Bcl-2, Bcl-xl and survivin were down-regulated, while Bax, AIF, cytochorome C and Smac/DIABLO were up-regulated in a dose-dependent manner. CONCLUSION Furanodienone can inhibite RKO cells proliferation and promote the RKO cells apoptosis, which may be one of the mecharisms to the activation of mitochondria-dependent pathway.
作者
江莹
密玉帅
王小琴
顾超
胡道德
JIANG Ying;MI Yu-shuai;WANG Xiao-qin;GU Chao;HU Dao-de(Department of Clinical Pharmacology;Department of General Surgery, the First People＇s Hospital, Shanghai Jiaotong University, SHANGHAI 200080, China)
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2018年第4期223-228,共6页
Chinese Journal of New Drugs and Clinical Remedies
基金
上海市科委基金项目(16401901400)
