氯喹增强LOVO细胞对5-氟尿嘧啶化疗敏感性的作用

目的研究结肠癌细胞LOVO在氯喹(CQ)作用下对化疗药物5-氟尿嘧啶(5-FU)的敏感性变化及机制。方法用MTT法分别检测CQ和5-FU作用24 h和48 h对LOVO细胞的增殖抑制作用。实验分为4组:空白对照组、CQ组、5-FU组和联合作用组。CQ、5-FU及两者联合作用于LOVO细胞后,通过细胞划痕实验和Transwell分别检测细胞迁移和侵袭的变化,采用流式细胞术和TUNEL方法检测细胞凋亡,采用Western blot检测细胞自噬及凋亡相关蛋白的表达情况。结果在不同浓度CQ、5-FU作用下LOVO细胞增殖受到抑制,且呈现剂量依赖性,48 h时CQ和5-FU的IC50分别为11.8μg/ml和2.5μmol/L。与空白对照组相比,CQ、5-FU作用后细胞迁移率和细胞侵袭能力显著降低,且两药联用组的抑制效应更显著。流式细胞术检测显示,CQ组、5-FU组细胞凋亡率分别为(17.85±1.04)%、(17.36±0.96)%,显著高于空白对照组(4.11±0.23)%,两药联用组凋亡率为(25.03±2.27)%,两药联用组与CQ、5-FU单用组相比,细胞凋亡率更高,差异均有统计学意义(P<0.01)。TUNEL检测显示,CQ组、5-FU组细胞凋亡指数显著高于空白对照组,两药联用组与CQ、5-FU单用组相比细胞核凋亡指数更高。Western blot检测显示CQ和5-FU处理后导致P53、Bax、caspase3、caspase9和P62蛋白表达水平显著升高,而Bcl-2、LC3和Beclin-1蛋白表达水平则显著降低,差异均有统计学意义(P<0.01)。结论 CQ能抑制结直肠癌细胞LOVO的增殖、迁移、侵袭和自噬,促进其凋亡,并能增强细胞对化疗药物5-FU的敏感性。

Chloroquine enhances the sensitivity of the LOVO cell line to the chemotherapeutic 5-fluorouracil

Objective To investigate the impact of chloroquine(CQ)on the sensitivity of the LOVO colon cancer cell line to 5-fluorouracil(5-FU)and the mechanism for that action.Methods An MTT assay was used to detect the inhibitory effect of CQ and/or 5-FU on LOVO cells 24 h and 48 h after treatment.Cell migration and invasion were assessed using a scratch assay and Transwell invasion assay,respectively,cell apoptosis was evaluated with flow cytometry and TUNEL,and the expression of autophagy-and apoptosis-related proteins was detected using Western blotting.Results The proliferation of LOVO cells was significantly inhibited by CQ and 5-FU in a dose-dependent manner.The IC50 of CQ at 48 h was 11.8 g/ml and that of 5-FU was 2.5 M.CQ and 5-FU significantly inhibited cell migration and invasion in comparison to the control group,and the inhibitory effect was more significant in cells exposed to both CQ and 5-FU.Flow cytometry indicated that the rate of apoptosis was 17.85il.04%in the CQ group and 17.36±0.96%in the 5-FU group.Rates were significantly higher than that in the control group(4.11±O.23%).The rate of apoptosis in the two-drug group was 25.03±2.27%,which was higher than that in the CQ and 5-FU groups.The difference in rates was significant(P<0.01).TUNEL indicated that both CQ and 5-FU significantly promoted apoptosis of LOVO cells,and the combination of CQ and 5-FU was more effective than either alone.The levels of P53,Bax,caspase3,caspase9,and P62 protein increased significantly while the levels of Bcl-2,LC3,and Beclin-1 decreased significantly with CQ and 5-FU treatment(P<0.01).Conclusion CQ inhibited the proliferation,migration,invasion,and autophagy of LOVO cells and promoted their apoptosis.Moreover,CQ enhanced the sensitivity of LOVO cells to the chemotherapy drug 5-FU.