摘要
目的：探究miR-567 对非小细胞肺癌（NSCLC）的生物学功能以及吉非替尼耐药性的影响。方法：采用RT-qPCR 检测miR-567 在人肺上皮细胞系BEAS-2B、NSCLC 细胞系（PC9、HCC827、A549、NCI-H1975）以及吉非替尼耐药细胞株PC9/GR 中的表达。PC9 细胞和PC9/GR 细胞分别分为negative control 组、miR-567 mimic 组、miR-567 inhibitor 组和miR-567 mimic+CDK8 组，CCK8 检测细胞的增殖能力，Transwell 法检测细胞迁移和侵袭能力，双荧光素酶报告基因实验检测miR-567 与CDK8 的靶向关系，western blotting 检测细胞中CDK8 的表达。结果：miR-567 在PC9、HCC827、A549、NCI-H1975 中的表达显著低于BEAS-2B，miR-567 在PC9/GR 细胞中的表达显著低于PC9。相比于negative control 组，miR-567 mimic 组PC9 细胞增殖、迁移和侵袭能力均显著减弱（P＜0.05）。miR-567 inhibitor 组PC9 细胞增殖、迁移和侵袭能力均显著增强（P＜0.05），miR-567 mimic 组PC9/GR 细胞增殖率显著低于negative control 组（P＜0.05）。miR-567 inhibitor 组PC9/GR 细胞增殖率显著高于negative control组（P＜0.05）。双荧光素酶报告实验和western blotting检测结果显示CDK8为miR-567的靶基因。相比于miR-567 mimic 组，miR-567 mimic+CDK8 组PC9 细胞的增殖、迁移以及侵袭能力均显著增强（P＜0.05），miR-567 mimic+CDK8 组PC9/GR 细胞增殖率显著升高（P＜0.01）。结论：miR-567 可通过靶向调控CDK8 而抑制NSCLC 细胞的增殖、迁移和侵袭能力以及吉非替尼耐药。
Abstract
Objective: To explore the effect of miR-567 on the biological function and gefitinib resistance of nonsmall cell lung cancer (NSCLC).Methods: Real-time fluorescence quantitative polymerase chain reaction (RTqPCR)was used to detect the expressions of miR-567 in human lung epithelial cell line BEAS-2B, non-small cell lung cancer cell line PC9, HCC827, A549, NCI-H1975, and gefitinib-resistant cell line PC9/GR.PC9 cells and PC9/GR cells were divided into negative control group, miR-567 mimic group, miR-567 inhibitor group and miR-567 mimic+CDK8 group, respectively.Cell proliferation ability was detected by CCK8 method, the migration and invasion ability of cells were detected by Transwell cell assay, the targeting relationship between miR-567 and CDK8 was detected by double luciferase reporter gene assay, and the expression of CDK8 in cells was detected by western blotting.Results: The expression of miR-567 in PC9, HCC827, A549 and NCI-H1975 was significantly lower than that in BEAS-2B, and the expression of miR-567 in PC9/GR cells was significantly lower than that in PC9.Compared with negative control group, proliferation, migration and invasion ability of PC9 cells in miR-567 mimic group significantly decreased (P＜0.05) and those in miR-567 inhibitor group significantly increased (P＜0.05).The proliferation rate of PC9/GR cells in miR-567 mimic group was significantly lower than that in negative control group (P＜0.05), and the proliferation rate of PC9/GR cells in miR-567 inhibitor group was significantly higher than that in negative control group (P＜0.05).Double luciferase assay and western blotting analysis showed that CDK8 was the target gene of miR-567.Compared with miR-567 mimic group, proliferation, migration and invasion ability of PC9 cells in miR-567 mimic+CDK8 group significantly increased (P＜0.05), and the proliferation rate of PC9/GR cells significantly increased (P＜0.01).Conclusion: miR-567 can inhibit proliferation, migration, invasion and gefitinib resistance of non-small cell lung cancer cells by targeting CDK8 regulation.
百度学术
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