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Open Access 01-12-2016 | Research

Overexpression of RBM5 induces autophagy in human lung adenocarcinoma cells

Authors: Zhenzhong Su, Ke Wang, Ranwei Li, Jinzhi Yin, Yuqiu Hao, Xuejiao Lv, Junyao Li, Lijing Zhao, Yanwei Du, Ping Li, Jie Zhang

Published in: World Journal of Surgical Oncology | Issue 1/2016

Abstract

Background

Dysfunctions in autophagy and apoptosis are closely interacted and play an important role in cancer development. RNA binding motif 5 (RBM5) is a tumor suppressor gene, which inhibits tumor cells’ growth and enhances chemosensitivity through inducing apoptosis in our previous studies. In this study, we investigated the relationship between RBM5 overexpression and autophagy in human lung adenocarcinoma cells.

Methods

Human lung adenocarcinoma cancer (A549) cells were cultured in vitro and were transiently transfected with a RBM5 expressing plasmid (GV287-RBM5) or plasmid with scrambled control sequence. RBM5 expression was determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Intracellular LC-3 I/II, Beclin-1, lysosome associated membrane protein-1 (LAMP1), Bcl-2, and NF-κB/p65 protein levels were detected by Western blot. Chemical staining with monodansylcadaverine (MDC) and acridine orange (AO) was applied to detect acidic vesicular organelles (AVOs). The ultrastructure changes were observed under transmission electron microscope (TEM). Then, transplanted tumor models of A549 cells on BALB/c nude mice were established and treated with the recombinant plasmids carried by attenuated Salmonella to induce RBM5 overexpression in tumor tissues. RBM5, LC-3, LAMP1, and Beclin1 expression was determined by immunohistochemistry staining in plasmids-treated A549 xenografts.

Results

Our study demonstrated that overexpression of RBM5 caused an increase in the autophagy-related proteins including LC3-I, LC3-II, LC3-II/LC3-I ratio, Beclin1, and LAMP1 in A549 cells. A large number of autophagosomes with double-membrane structure and AVOs were detected in the cytoplasm of A549 cells transfected with GV287-RBM5 at 24 h. We observed that the protein level of NF-κB/P65 was increased and the protein level of Bcl-2 decreased by RBM5 overexpression. Furthermore, treatment with an autophagy inhibitor, 3-MA, enhanced RBM5-induced cell death and chemosensitivity in A549 cells. Furthermore, we successfully established the lung adenocarcinoma animal model using A549 cells. Overexpression of RBM5 enhanced the LC-3, LAMP1, and Beclin1 expression in the A549 xenografts.

Conclusions

Our findings showed for the first time that RBM5 overexpression induced autophagy in human lung adenocarcinoma cells, which might be driven by upregulation of Beclin1, NF-κB/P65, and downregulation of Bcl-2. RBM5-enhanced autophagy acts in a cytoprotective way and inhibition of autophagy may improve the anti-tumor efficacy of RBM5 in lung cancer.

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Title: Overexpression of RBM5 induces autophagy in human lung adenocarcinoma cells
Authors: Zhenzhong Su
Ke Wang
Ranwei Li
Jinzhi Yin
Yuqiu Hao
Xuejiao Lv
Junyao Li
Lijing Zhao
Yanwei Du
Ping Li
Jie Zhang
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: World Journal of Surgical Oncology / Issue 1/2016
Electronic ISSN: 1477-7819
DOI: https://doi.org/10.1186/s12957-016-0815-7

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Overexpression of RBM5 induces autophagy in human lung adenocarcinoma cells

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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