牛乳铁蛋白抗菌肽抗肺癌细胞增殖及其作用机制研究

Functions and mechanism of bovine lactoferricin antimicrobial peptide’s resistance to lung cancer cells

目的探讨牛乳铁蛋白抗菌肽(LfcinB)抗人肺癌细胞功能及其作用机制。方法体外培养人肺腺癌A549细胞株,实验分为:阴性对照组、LfcinB处理组和阳性对照组。采用MTT法检测LfcinB对人肺腺癌A549细胞增殖的影响,流式细胞仪测定细胞周期和凋亡率。结果与阴性对照组比较,不同浓度的LfcinB均对A549细胞增殖有抑制作用,其中以10-2g/L浓度LfcinB的抑制率最高(P<0.01)。与阴性对照组比较,不同浓度的LfcinB作用于A549细胞24、48、72 h后,G0/G1期细胞略有减少,S期细胞逐渐增多,G2/M期细胞略有减少,且在72 h结果最为显著(P<0.01)。Lf-cinB在10-6～10-2g/L浓度范围内作用48、72 h均可诱导A549细胞凋亡,随着药物浓度增高,诱导凋亡作用加重,有明显的剂量效应关系。结论 LfcinB能显著抑制人肺腺癌A549细胞增殖,使细胞阻滞于S期,并诱导细胞凋亡。

Objective To observe the function and mechanism of bovine lactoferricin antimicrobial peptides （Lf- cinB） on the treatment of human lung cancer cell. Methods After culture of human lung adenocarcinoma cells （A549） with medium, experiments were divided into： negative control group, LfcinB group and positive control groups. The MTT assays were used to detect the cell proliferation of bovine lactoferrin antimicrobial peptides against human lung adenocarcinoma cell. Apotosis and cycles of the cells were detected by flow cytomentry with annexin V FITC/PI double staining. Results LfcinB treatment with different concentrations all has effect on inhibiting the proliferation of A549 cell compared with the negative control group. The inhibition rate of 10-2 g,/L LfcinB was the highest（P 〈 0. 01 ）. In comparison to A549 cell the negative control group,The treatment of LfcinB with different concentrations for 24,48,72 h slightly decreased the cell counts in GO - G1 phase and G2 - M phase, while it grad- ually increased the cells in S phase. It was significantly higher in the group for 72 h than the others（ P 〈0. 01 ）. Lactoferrin antimicrobial peptides treatment with different concentrations for 48,72 h could induce the apotosis of A549 cell in a dose dependent manner. Conclusion LfcinB can significantly restrain human lung adenocarcinoma cell line A549 proliferation, induct cell apoptosis and hinder cell generation cycle in S phase.