摘要
目的:探讨细胞自噬对肝癌Bel-7402/FU细胞5-氟尿嘧啶敏感性的影响。方法:选取Bel-7402、Bel-7402/FU细胞株,分为对照组、5-氟尿嘧啶组、自噬抑制剂3-甲基腺苷组以及5-氟尿嘧啶+3-甲基腺苷组,MTT法了解抑制细胞自噬对肝癌5-氟尿嘧啶IC50值的影响;流式细胞术检测抑制细胞自噬对细胞凋亡的影响;GFP-LC3质粒转染观察细胞浆中GFP-LC3分布情况。结果:Bel-7402细胞株与Bel-7402/FU细胞株5-氟尿嘧啶IC50值分别为4.66、68.14μg/mL,凋亡率分别为13.80%、1.09%。3-甲基腺苷联合5-氟尿嘧啶作用后Bel-7402细胞株与Bel-7402/FU细胞株的氟尿嘧啶IC50值分别为4.31、29.44μg/mL,凋亡率分别为13.82%、6.86%。在3-甲基腺苷作用下,Bel-7402/5-FU细胞株5-氟尿嘧啶诱导的点状样GFP-LC3的细胞数量减少。结论:抑制细胞自噬可降低Bel-7402/FU对5-氟尿嘧啶IC50值,诱导细胞凋亡,从而有效地逆转Bel-7402/FU对5-氟尿嘧啶耐药。
AIM: To explore the effects of autophagy in the sensitivity of Hepatocellular carcinoma cells line Bel-7402/FU to 5-FU. METHODS: Hepatocellular carcinoma cells lines Bel-7402 and Bel-7402/FU were treated with 5-FU or 3-MA, or pretreated with 3-MA followed by 5-FU. MTT method test was used to determine the proliferative inhibition rate. Flow cytometry was used to examine cell apoptosis. The GFPLC3B morphological changes were observed by fluorescent microscope. RESULTS: The IC50 values of 5-FU in Bel-7402 and Bel-7402/FU cells were 4.66 μg /mL and 68.14 μg /mL, respectively. The apoptosis rates of the Bel-7402 and Bel-7402/FU were 13.80% and 1. 09%, respectively. When 5-FU was used combined with 3- MA, the IC50 values of 5-FU in Bel-7402 and Bel-7402/FU cells were 4.31 μg /mL and 29.44 μg /mL. The apoptosis rates of Bel-7402 and Bel-7402/FU were 13.82% and 6.86%, respectively. 3-MA decreased 5-FU-induced punctuate spot formation in Bel-7402/FU. CONCLUSION: Autophagy is connected with 5-FU-resistance and inhibition of autophagy can reverse 5-FU-resistance in Bel-7402 /FU.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2014年第5期488-492,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
安徽省自然科学基金(11040606M222)