抗菌肽17BIPHE2对人肺腺癌细胞A549的抑制作用

李军; 回丽媛; 战仕胜; 王秀青

doi:10.3971/j.issn.1000-8578.2017.17.0287

抗菌肽17BIPHE2对人肺腺癌细胞A549的抑制作用

Inhibition of Antimicrobial Peptide 17BIPHE2 on Lung Adenocarcinoma A549 Cells and Related Mechanism

摘要

摘要:
目的探讨抗菌肽17BIPHE2对人肺腺癌细胞A549的抑制作用及其作用机制。
方法 MTT法检测0~40 μmol/L的抗菌肽17BIPHE2对A549细胞增殖的影响；流式细胞术和TUNEL染色检测细胞凋亡情况；Real-time PCR和Western blot检测Bax、Bcl-2的表达变化；透射电子显微镜观察细胞超微结构变化。
结果 17BIPHE2可抑制A549细胞的增殖，其IC₅₀值为34.33 μmol/L，同时20、25、30、35、40 μmol/L的17BIPHE2对A549细胞的生长抑制率分别为（1.8±2.7）%、（6.1±2.1）%、（24.6±4.6）%、（55.2±1.1）%、（76.3±1.2）%。与对照组相比，25、35 μmol/L的17BIPHE2处理24h的A549细胞凋亡率显著增高（P < 0.05），细胞核呈现固缩，染色质深染，线粒体嵴断裂，内质网肿胀等凋亡细胞的特征。Bax的表达上调（P < 0.05），Bcl-2的表达受到抑制（P < 0.05），Bax/Bcl-2比值升高。
结论 17BIPHE2可能是通过上调Bax表达，下调Bcl-2表达诱导A549细胞凋亡，抑制其增殖。

Abstract:
Objective To investigate the inhibition of antimicrobial peptide 17BIPHE2 on lung adenocarcinoma A549 cells and the related mechanism.
Methods The viabilities of A549 cells treated with 17BIPHE2(0-40μmol/L) were measured by MTT method; flow cytometry and TUNEL were used to observe cell apoptosis; Real-time PCR and Western blot were used to measure the expression of Bax and Bcl-2. Transmission electron microscope was used to observe the ultrastructure change of A549 cells.
Results MTT results showed that 17BIPHE2 could inhibit the proliferation of A549 cells, and the IC₅₀ was 34.33μmol/L. The inhibitory rates at different concentrations (20, 25, 30, 35 and 40μmol/L) of 17BIPHE2 was (1.8±2.7)%, (6.1±2.1)%, (24.6±4.6)%, (55.2±1.1)% and (76.3±1.2)%, respectively. The apoptosis rates of A549 cells treated with 17BIPHE2 (25, 35μmol/L) for 24h were significantly higher than that of the control group(P < 0.05); the nucleus of A549 presented karyopyknosis, hyperchromatin mitochondrial crest fracture, endoplasmic reticulum swelling and other characteristics of apoptosis. Real-time PCR and Western blot results showed that 17BIPHE2 significantly upregulated Bax expression(P < 0.05) and downregulated Bcl-2 expression(P < 0.05) respectively, and the ratio of Bax/Bcl-2 was upregulated.
Conclusion Antimicrobial peptide 17BIPHE2 may induce the apoptosis and suppress the proliferation of A549 cells by upregulating Bax expression and downregulating Bcl-2 expression.

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