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短发夹RNA介导的α-N-乙酰半乳糖胺酶下调抑制癌细胞系的迁移和侵袭。

shRNA-mediated downregulation of α-N-Acetylgalactosaminidase inhibits migration and invasion of cancer cell lines.

作者信息

Saburi Ehsan, Tavakolafshari Jalil, Mortazavi Yousef, Biglari Alireza, Mirzaei Seyed Abbas, Nadri Samad

机构信息

Department of Molecular Medicine & Genetics, Zanjan University of Medical Sciences, Zanjan, Iran.

Department of Immunogenetics, Buali Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

出版信息

Iran J Basic Med Sci. 2017 Sep;20(9):1021-1028. doi: 10.22038/IJBMS.2017.9271.

Abstract

OBJECTIVES

Extracellular matrix (ECM) is composed of many kinds of glycoproteins containing glycosaminoglycans (GAGs) moiety. The research was conducted based on the N-Acetylgalactosamine (GalNAc) degradation of ECM components by α-N-acetylgalactosaminidase (Nagalase) which facilitates migration and invasion of cancer cells. This study aims to investigate the effects of Naga-shRNA downregulation on migration and invasion of cancer cell lines.

MATERIALS AND METHODS

In this study, MCF-7 cell line (human mammary carcinoma cell line) and A2780 (human ovarian carcinoma cell line) were used. The level of normalized Naga expression and Nagalase protein were evaluated by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay/western blotting, respectively. Migration and invasion were determined using transwell assays, and statistical analysis was carried out by ANOVA test.

RESULTS

Response to transduction by shRNA compared to the control group, migrative and invasive properties of the transfected cells were significantly inhibited.

CONCLUSION

These results indicate that Nagalase may have an important role in migration and invasion of cancer cells and can be considered as a candidate for further studies.

摘要

目的

细胞外基质(ECM)由多种含有糖胺聚糖(GAGs)部分的糖蛋白组成。该研究基于α-N-乙酰半乳糖胺酶(Nagalase)对ECM成分的N-乙酰半乳糖胺(GalNAc)降解展开,这种降解促进癌细胞的迁移和侵袭。本研究旨在探究Naga-shRNA下调对癌细胞系迁移和侵袭的影响。

材料与方法

本研究使用了MCF-7细胞系(人乳腺癌细胞系)和A2780(人卵巢癌细胞系)。分别通过定量聚合酶链反应和酶联免疫吸附测定/蛋白质印迹法评估标准化的Naga表达水平和Nagalase蛋白水平。使用Transwell实验测定迁移和侵袭情况,并通过方差分析进行统计分析。

结果

与对照组相比,shRNA转导后的反应显示,转染细胞的迁移和侵袭特性受到显著抑制。

结论

这些结果表明,Nagalase可能在癌细胞的迁移和侵袭中起重要作用,可被视为进一步研究的候选对象。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2034/5651455/1962960e48a4/IJBMS-20-1021-g001.jpg

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