miR-483-3p靶向CD44抑制EGFR突变肺癌的作用研究

目的:探讨mi R-483-3p对CD44表达的调控作用及采用脂质体载药系统递送mi R-483-3p治疗表皮生长因子(epidermalgrowth factor receptor,EGFR)突变的非小细胞肺癌(non-small cell lung cancer,NSCLC)的作用。方法:通过mi R-483-3p靶基因的数据库发现CD44可能是mi R-483-3p的靶基因之一,从结构上及功能上进行验证。在EGFR酪氨酸激酶抑制剂(epidermal growth factor receptor-tyrosine kinase inhibitor, EGFR-TKI)耐药的NSCLC模型中,分别从基因及蛋白水平检测耐药模型及敏感模型中CD44的表达。在HCC827GR移植瘤模型中,采用脂质体-鱼精蛋白-DNA (liposome-polycation-DNA, LPD)载药系统递送mi R-483-3p进行治疗,观察肿瘤的生长情况。结果:双荧光素酶报告基因实验及Western blot实验结果显示CD44是mi R-483-3p的靶基因之一,且CD44在EGFR-TKI耐药模型中异常高表达(P0.05)。脂质体载药系统LPD-mi RNA-DSPE-PEG符合静脉给药要求,Size为66.93±21 nm,Zeta potential为8.7±2 m V,PDI (Polydispersity Index)为0.1,递送mi R-483-3p后能够抑制HCC827GR移植瘤的生长(P0.05)。结论:CD44为mi R-483-3p的靶基因之一,在耐药模型中高表达,脂质体载药系统给予mi R-483-3p能够抑制EGFR-TKI耐药肿瘤的生长。

Inhibitory Effect of miR-483-3p on the EGFR-mutant Lung Cancer Via Targeting CD44

ABSTRACT Objective: To investigate the regulation of miR-483-3p on CD44 and the treatment of miR-483-3p in EGFR (epidermal growth factor receptor) mutations non-small cell lung cancer (NSCLC) by liposome drug delivery system. Methods: CD44 was found to be one of the target genes of miR-483-3p in the database, and was verified structurally and functionally. In the EGFR Tyrosine Kinase Inhibitor (EGFR-TKI)-resistant model of NSCLC, the expression of CD44 which was detected from gene and protein levels in the resistant and sensitive models. In the xenograft model of HCC827GR, miR-483-3p was delivered for treatment by the liposome- protamine-DNA (LPD) drug delivery system to observe the growth of tumor. Results: Dual-Luciferase Reporter Assay System and Western Blot showed that CD44 was one of the target genes of miR-483-3p, and CD44 was abnormally highly expressed in the EGFR-TKI resistant model in vitro and in vivo(P<0.05). The LPD-miRNA-DSPE-PEG prepared by the liposome drug-loading system met the requirements of intravenous administration, Size: 66.93±21 nm, Zeta potential: 8.7±2 mV, Polydispersity Index: 0.1, which could inhibit the growth of HCC827GR-xenograft-model after delivery of miR-483-3p by LPD-miRNA-DSPE-PEG(P<0.05). Conclusion: CD44 is one of the target genes of miR-483-3p and is highly expressed in the resistant model. The liposome drug delivery system about miR-483-3p can inhibit the growth of EGFR-TKI resistant tumors.