内吞适配蛋白Epsin在非小细胞肺癌发生中的作用研究

The Potential Role of Endocytic Adaptor Protein Epsin in the Development of Non-small Cell Lung Cancer

目的:探讨内吞适配蛋白Epsin在非小细胞肺癌发生中的潜在作用。方法:选择体外培养的人非小细胞肺癌细胞(A549),筛选Epsin 1和Epsin 2 shRNA干扰效率达标的细胞。将裸鼠随机分为3组,每组10只,第1、2组裸鼠分别经胸腔植入人非小细胞肺癌细胞(A549)及epsin表达敲减的A549细胞,第3组注射等量的生理盐水,比较1、2组小鼠肿瘤体积的变化。8周后,处死所有裸鼠,留取肺组织及肿瘤组织,通过免疫荧光染色检测非肿瘤(正常)肺和致瘤性肺组织中的epsin 1和2的蛋白质水平。用实时定量PCR(qRT-PCR)来研究epsin 1和2的基因表达水平。结果:肺肿瘤组织epsin1和2的m RNA和蛋白表达均显著高于正常肺组织中(P<0.05)。种植epsin表达敲减的A549细胞裸鼠肿瘤生长速度及体积均大于种植正常A549细胞的裸鼠肿瘤。结论:Epsins表达上调可能促进非小细胞肺癌肿瘤的发生发展,而敲减epsins的表达可能为未来的非小细胞肺癌的治疗提供新的治疗靶点。

Objective: To explore the potential role of endocytic adaptor protein Epsin in the development of non-small cell lung cancer. Methods: Human non-small cell lung cancer cells(A549) cultured in vitro were selected and screened for Epsin 1 and Epsin 2 shRNA interference efficiency. Nude mice were randomly divided into 3 groups, 10 in each group. The first and second groups of nude mice were implanted into human non-small cell lung cancer cells(A549) and epsilon-expressed A549 cells by thoracic cavity, and the third group was injected with the same amount. The normal saline was compared to the changes in tumor volume of mice in groups 1 and2. Eight weeks later, all nude mice were sacrificed, lung tissue and tumor tissues were taken, and protein levels of epsin 1 and 2 in non-tumor(normal) lung and tumorigenic lung tissues were detected by immunofluorescence staining. Real-time quantitative PCR(qRT-PCR) was used to study the gene expression levels of epsin 1 and 2. Results: The m RNA and protein expressions of epsin1 and 2 in lung tumor tissues were significantly higher than those in normal lung tissues(P<0.05). The tumor growth rate and volume of A549 cells implanted with epsin expression knockdown were larger than those of nude mice implanted with normal A549 cells. Conclusions:Up-regulation of Epsins may promote the development of non-small cell lung cancer, and knocking down the expression of epsins may provide a new therapeutic target for the treatment of non-small cell lung cancer in the future.