Inhibition of Antimicrobial Peptide 17BIPHE2 on Lung Adenocarcinoma A549 Cells and Related Mechanism
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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。
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关键词:
- 抗菌肽17BIPHE2 /
- 肺腺癌 /
- A549细胞 /
- 凋亡
Abstract:ObjectiveTo investigate the inhibition of antimicrobial peptide 17BIPHE2 on lung adenocarcinoma A549 cells and the related mechanism.
MethodsThe 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.
ResultsMTT 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.
ConclusionAntimicrobial 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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Key words:
- Antimicrobial peptide 17BIPHE2 /
- Lung adenocarcinoma /
- A549 cells /
- Apoptosis
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图 1 抗菌肽17BIPHE2和紫杉醇抑制A549细胞增殖
Figure 1 17BIPHE2 and paclitaxel inhibited proliferation of A549 cells
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图 2 流式细胞术检测不同浓度17BIPHE2对A549细胞凋亡的影响
Figure 2 Apoptosis rates of A549 cells treated with different concentrations of 17BIPHE2 observed by flow cytometry
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图 3 TUNEL染色检测17BIPHE2诱导的A549细胞凋亡
Figure 3 Apoptosis of A549 cells induced by 17BIPHE2 observed by TUNEL staining assay
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图 4 17BIPHE2对A549细胞Bax、Bcl-2 mRNA表达的影响
Figure 4 Effect of 17BIPHE2 on Bax mRNA and Bcl-2 mRNA expression in A549 cells
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图 5 17BIPHE2对A549细胞Bax、Bcl-2蛋白表达的影响
Figure 5 Effect of 17BIPHE2 on Bax and Bcl-2 protein expression in A549 cells
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图 6 透射电子显微镜观察17BIPHE2对A549细胞超微结构的影响
Figure 6 Effect of 17BIPHE2 on ultrastructure of A549 cells observed by transmission electron microscopy
表 1 不同浓度17BIPHE2处理A549细胞24 h后的细胞凋亡率(n=3, x±s, %)
Table 1 Cell apoptosis rates of A549 cells treated with different concentrations of 17BIPHE2 for 24 h (n=3, x±s, %)
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