摘要
目的:探讨解毒三根汤联合5-FU对人结肠癌耐药细胞株LOVO/5-FU的逆转耐药作用,明确糖酵解在肿瘤耐药逆转中的作用。方法:采用药物持续接触浓度递增法建立LOVO/5-FU耐药细胞株。CCK-8法检测其耐药性,测定不同浓度解毒三根汤单用及与5-FU联用对细胞增殖抑制率的影响。Western Blot法检测PI3K/AKT/HIF-1α通路和caspase3表达量。结果:成功建立人结肠癌LOVO/5-FU耐药细胞株,IC50为(287.05±59.18)μg/mL,耐药倍数为43。解毒三根汤能明显抑制LOVO/5-FU细胞株的耐药性,并呈剂量依赖性。Western Blot显示解毒三根汤作用后,协同组的caspase3表达量有所增加,HIF-1α、AKT、P-AKT、PI3K、Glut1、HKⅡ蛋白表达量显著减少(P<0.05)。结论:解毒三根汤能显著逆转人结肠癌耐药细胞株LOVO/5-FU的耐药性,其机制可能与抑制PI3K/AKT通路的活化,诱导细胞凋亡,下调HIF-1α从而调控糖酵解实现的。
Objective:To investigate the reversal effects of Jiedu Sangen Decoction combined with 5-FU on drugresistant human colon cancer cell line LOVO/5-FU,and clarify the effect of glycolysis in reversing drug-resistant.Methods:Drug resistant human colon cancer cell line LOVO/5-FU was induced by continuously exposing of gradually increasing concentrations of 5-FU.Using CCK-8 assay to detect drug-resistance,and determine the cell growth inhibiting ratio of Jiedu Sangen Decoction in different concentrations alone and combined with concentration gradient of 5-FU in LOVO/5-FU cell line.The expression level of PI3 K/AKT/HIF-1αsignal pathway and caspase3 were detected by Western Blot.Results:Drug-resistant human colon cancer cell line LOVO/5-FU was successfully established.The half inhibitive concentration of LOVO/5-FU was(287.05±59.18)μg/mL,the resistance index(RI)was 43.Jiedu Sangen Decoction can significantly inhibit LOVO/5-FU cell line drug-resistance in a dose-dependent manner.Western Blot showed the caspase3 protein in collaborative group increased while the protein of HIF-1α,AKT,P-AKT,PI3 K,Glut1,HK II decreased,with statistical significance(P<0.05).Conclusion:Jiedu Sangen Decoction has a significantly effect on reversing the drug-resistance in human colon cancer cell line LOVO/5-FU.The reversing effect of Jiedu Sangen Decoction may be achieved by inhibiting the activation of PI3 K/AKT pathway,inducing apoptosis and reducing the expression of HIF-1αto regulate glycolysis.
作者
严卿莹
阮善明
张恺
孙磊涛
周萌萌
谢璐帆
洪雨心
沈敏鹤
YAN Qing-ying;RUAN Shan-ming;ZHANG Kai;SUN Lei-tao;ZHOU Meng-meng;XIE Lu-fan;HONG Yu-xin;SHEN Min-he(The First Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,China;The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310000,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2019年第4期1751-1756,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
浙江省中医药科技计划重点研究项目(No.2015Z013)~~
