CpG-ODN对肺癌A549细胞化疗的协同作用

CpG-ODN enhances sensitivity to chemotherapy in lung cancer cell line A549

关键词:  非小细胞肺癌  CpG寡聚脱氧核苷酸  化学疗法

Key words:  non-small cell lung cancer  CpG-ODN  chemotherapy

摘要:

目的 研究CpG寡聚脱氧核苷酸(CpG-ODN)对肺癌A549细胞化疗的影响,并探讨其可能的作用机制。方法 将肺癌A549细胞分为空白对照组、DDP组、CpG-ODN组、DDP+CpG-ODN组、CpG-ODN+DDP组。采用CCK-8比色法检测细胞的增殖情况,流式细胞仪检测细胞凋亡情况,RT-PCR检测TLR9 mRNA的表达,Western印迹法检测TLR9信号通路中MyD88、AP-1的表达。结果 TLR9激活CpG ODN后,能显著提高细胞的增殖能力(P<0.05)；CpG-ODN与CpG-ODN+DDP两组细胞生长无明显差异(P>0.05)；CpG-ODN+DDP组细胞凋亡率较DDP组低(P<0.05),DDP+CpG-ODN组细胞凋亡率较DDP组明显升高(P<0.05)；DDP+CpG-ODN组TLR9 mRNA较DDP组明显升高(P<0.05)；DDP+CpG组AP-1蛋白表达上调(P<0.05)。结论 CpG-ODN激活TLR9信号通路后,可协同DDP促进细胞凋亡,提高顺铂化疗的敏感性。

Abstract:

Objective To investigate the effect of TLR9 activator CpG-ODN on chemosensitivity of lung cancer A549 cells. Methods Cultured lung cancer A549 cells were divided into control group, DDP group, CpG-ODN group, DDP+CpG-ODN group, CpG-ODN+DDP group. Cell proliferation was analyzed with CCK-8 method; cell apoptosis was examined with flow cytometry; the mRNA expression of TLR9 was detected with RT-PCR; and the protein expression of MyD88 and AP-1 was detected with Western blotting. Results The cell proliferation was improved in CpG-ODN groups(P<0.05); there was no significant difference between CpG-ODN and CpG-ODN+DDP groups(P>0.05). The apoptosis rate of CpG-ODN+DDP was lower than that of DDP(P<0.05), and the apoptotic rate of DDP+CpG-ODN was significantly higher than that of DDP(P<0.05). The expression of TLR9 mRNA in DDP+CpG-ODN was higher than that in DDP group(P<0.05). The expression of AP-1 in DDP+CpG group was up-regulated(P<0.05). Conclusion The data indicate that the CpG-ODN can increase cell apoptosis and enhance the sensitivity of lung cancer cells to cisplatin chemotherapy.