Earle’s 盐平衡液饥饿诱导 DLD-1，HCT-116，A2780，CHO，Hep G2和 SMMC7721肿瘤细胞系自噬发生的时间优化

摘要

Autophagy has emerged as a key effector in tumorgenesis,but its precise function remains elusive.Au-tophagy 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 consid-erate 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 toler-ance 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.%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、HepG2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 HepG2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。