中文摘要：

肿瘤细胞在正常培养条件下自噬水平非常低,很难被监测到。与药物性自噬诱导剂相比,Earle＇s盐平衡液（EBSS）应用简便,除产生饥饿诱导自噬外,无其他细胞毒性,有利于研究细胞自噬的分子机制和功能。本文应用蛋白免疫印记和细胞免疫荧光技术探讨了EBSS对DLD-1和HCT-116结直肠癌、A2780和CHO卵巢癌、HepG 2和SMMC7721肝癌细胞系的自噬诱导效果和有效诱导时间。结果显示：EBSS短短几小时处理,能够非常有效地诱导上述6株肿瘤细胞发生自噬,但不同细胞系之间的有效诱导时间却明显不同。其中,DLD-1、HCT-116、CHO和HepG 2细胞系的有效诱导时间仅需2 h;A2780细胞需要稍微延长到4 h;然而SMMC7721却要延长到8 h左右。本研究为其他研究者在应用EBSS诱导肿瘤细胞自噬时提供了实践依据和参考。

英文摘要：

Autophagy has emerged as a key effector in tumorgenesis,but its precise function remains elusive. Autophagy flux is too low to be detected in normal culture conditions. Earle＇s balanced salts solution（ EBSS） is often used to induce autophage with two distinct advantages compared with other inducers： one is only needed to considerate the induction time,rather than the drug concentration; the other is that it exerts only starvation pressure by specifically simulating amino acids deprivation to cells without other undistinguishable cytotoxicity. Given the tolerance threshold of outside stimulus differs in various types of tumor cells,in this study we carried out the autophage induction experiments with several time points in DLD- 1 and HCT- 116 colon cancer cell lines,A2780 and CHO ovarian cancer cell lines,Hep G2 and SMMC7721 hepatocellular carcinoma cell lines. Our western-blotting and Immunofluorescence staining results demonstrated that EBSS can efficiently induce autophagy in the assayed cells,precisely with 2 hours treatment in DLD- 1,HCT- 116,Hep G2 and CHO cells,4 hours in A2780 and 8 hours in SMMC7721 cells,respectively. Our results provide a reference for autophagy induction with EBSS in cancer cells,and would be useful to shorten the time-consuming optimizing process.