3-MA通过抑制自噬反应提高结肠癌5-FU的化疗效果

目的 探索新的联合化疗方案以提高5-FU为基础的结肠癌化疗效果。方法 研究5-FU（7.5μmol/L）、3-MA（5mmol/L）以及两种药物联合使用对人结肠癌细胞株HCT116的自噬及凋亡影响。MTT、Hoechst＋PI染色检测细胞生长及凋亡,MDC染色观察细胞内自噬反应,Western blotting观察细胞内的凋亡及自噬相关蛋白改变。将HCT116细胞注射至裸鼠皮下建立在体肿瘤模型并进行治疗,观察肿瘤生长情况。结果MTT检测结果显示HCT116在5-FU作用下细胞生长受到抑制,Hoechst＋PI染色显示5-FU联合3-MA组凋亡数量较5-FU组明显增加;Western blott...

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Published in西安交通大学学报（医学版） Vol. 37; no. 1; pp. 49 - 53
Main Author 侯妮 刘娜 韩佳 李杰
Format Journal Article
LanguageChinese
Published 西安交通大学 医学部细胞生物学与遗传学系,陕西西安,710061%西安交通大学 第二附属医院普通外科,陕西西安,710004 2016
Subjects
3-MA
5-FU
Caspase-3
LC3
PARP
凋亡
化疗
结直肠癌
自噬
colorectal cancer
autophagy
apoptosis
PARP
化疗
Caspase-3
chemotherapy
自噬
结直肠癌
3-MA
凋亡
LC3
5-FU
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Summary:目的 探索新的联合化疗方案以提高5-FU为基础的结肠癌化疗效果。方法 研究5-FU（7.5μmol/L）、3-MA（5mmol/L）以及两种药物联合使用对人结肠癌细胞株HCT116的自噬及凋亡影响。MTT、Hoechst＋PI染色检测细胞生长及凋亡,MDC染色观察细胞内自噬反应,Western blotting观察细胞内的凋亡及自噬相关蛋白改变。将HCT116细胞注射至裸鼠皮下建立在体肿瘤模型并进行治疗,观察肿瘤生长情况。结果MTT检测结果显示HCT116在5-FU作用下细胞生长受到抑制,Hoechst＋PI染色显示5-FU联合3-MA组凋亡数量较5-FU组明显增加;Western blotting检测5-FU联合3-MA组的凋亡蛋白Caspase-3、PARP表达较5-FU组明显增高。同时,MDC染色以及自噬特异性蛋白LC3II检测发现在5-FU作用下自噬反应升高,而通过3-MA抑制5-FU引起的细胞自噬反应,细胞生长抑制率从5-FU单独作用的（51.64±4.04）%提高至（79.89±1.35）%,凋亡增加超过100%。皮下注射HCT116细胞的裸鼠肿瘤模型,5-FU联合3-MA组较单独使用5-FU组有效抑制肿瘤生长,肿瘤体积及质量均明显减少。结论 3-MA可抑制5-FU处理引起的细胞自噬反应,促进细胞凋亡,HCT116细胞生长抑制率明显增加;为提高5-FU为基础的结肠癌化疗提供新思路。
Bibliography:Objective To explore a novel adjuvant strategy of improving 5-fluorouracil （5-FU）-based chemotherapy for colorectal cancer. Methods Human colon cancer cell （HCT 116） was used to investigate the effect of 5-FU （7.5 μmol/L）, 3-methyladenine （3-MA, 5 mmol/L）, or their combination on apoptotic cell death and autophagy. MTT assay was used to observe the suppression of cell survival. Hoechst plus PI staining and MDC assay was used to detect apoptosis and autophagy. Western blotting was used to explore the molecular changes. Finally, the experiment results were further examined in vivo. Results We found that the survival of HCT116 was suppressed by 5-FU and the combination of 5-FU and 3-MA promoted the apoptosis. Autophagy was activated by 5-FU treatment and 3-MA inhibited 5-FU-induced autophagy. The suppression of cell survival increased from （51.64±4.04）% to （79.89±1.35）%, and the apoptosis increased by more than 100%. HCT116 cells were injected in the nude mouse to produce tumor. The combination treatment of 5-FU
ISSN:1671-8259
DOI:10.7652/jdyxb201601009