背景:以前,AF-956,其中包含FAM83G的S356和用于进入结肠癌细胞的N末端天线肽,与对照肽(AF-859)相比,具有明显的抗增殖作用,缺少N端天线肽,通过抑制残基S15和S82处的HSP27磷酸化来诱导细胞凋亡。
目的:因为FAM83G衍生肽是结肠癌治疗的有前景的先导化合物,我们重新分析了AG-066的作用,它含有FAM83G的S356和一个N端触角肽进入肝癌细胞.
方法:将HepG2肝癌细胞与浓度为54μM的AF-859或AG-066在37°C下孵育24、48和72小时。使用倒置光学显微镜评估AF-859和AG-066对培养的HepG2细胞的影响。此外,通过应用活/死细胞染色试剂盒II进行DNA梯法和死细胞测定。通过蛋白质印迹估计Erk磷酸化。
结果:与AF-859处理的HepG2细胞相比,用AG-066处理的HepG2活细胞计数显著减少,从培养基中死亡细胞数量的显著增加可以明显看出。此外,AG-066处理增加了细胞DNA阶梯。我们发现AG-066和AF-859治疗组之间的Erk磷酸化状态没有差异。
结论:这项研究表明,具有基于FAM83G结构的肽可作为肝癌细胞的自发凋亡诱导剂。因此,它是治疗肝癌的一种有前途的先导化合物。
BACKGROUND: Previously, AF-956, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into colon cancer cells, is markedly antiproliferative compared to a control peptide (AF-859), which lacks the N-terminal antenna peptide, by inducing apoptosis via the inhibition of HSP27 phosphorylation at residues S15 and S82.
OBJECTIVE: Because FAM83G-derived peptides are promising lead compounds for colon cancer treatment, we reanalyzed the effect of AG-066, which contains S356 of FAM83G and an N-terminal antenna peptide for entry into the liver cancer cells.
METHODS: HepG2 liver cancer cells were incubated with either AF-859 or AG-066 at a concentration of 54 μM at 37 °C for 24, 48, and 72 h. The effects of AF-859 and AG-066 on the cultured HepG2 cells were estimated using an inverted light microscope. Furthermore, the DNA ladder method and the dead cell assay were performed by applying Live/Dead Cell Staining Kit II. Erk phosphorylation was estimated by western blotting.
RESULTS: Treatment with AG-066 markedly reduced HepG2 viable cell counts compared to the AF- 859-treated HepG2 cells, as evident from the significantly increased number of dead cells in the culture medium. Additionally, AG-066 treatment increased cellular DNA laddering. We found no difference in Erk phosphorylation status between the AG-066- and AF-859-treated groups.
CONCLUSIONS: This study illustrated that the peptide with a structure based on FAM83G functions as a spontaneous apoptosis inducer for liver cancer cells. Hence, it is a promising lead compound for the treatment of liver cancer.