Lentiviral vector-mediated RBM5 overexpression downregulates EGFR expression in human non-small cell lung cancer cells

Su, Zheng; Yin, Jinzhi; Zhao, Lijing; Li, Ranwei; Hong, Liang; Zhang, Jie; Wang, Ke

doi:10.1186/1477-7819-12-367

Cited by 10 publications

(12 citation statements)

References 39 publications

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“…Besides the above two mechanisms, other signaling may be also involved in the anti-tumor role of RBM5. For example, RBM5 was also shown to downregulate EGFR expression and induce cell autophagy [20, 21]. Autophagy is another autonomic programmed cell death or type II PCD.…”

Section: Discussionmentioning

confidence: 99%

RBM5 inhibits tumorigenesis of gliomas through inhibition of Wnt/β-catenin signaling and induction of apoptosis

et al. 2017

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BackgroundGliomas are one of the most common malignant brain tumors and bring a big threat to human life as traditional therapy is unsatisfactory. RBM5 was a RNA-binding motif protein and was reported as a tumor suppressor. But the role of RBM5 in gliomas was unknown.MethodsThe mRNA level of RBM5 was determined in gliomas tissues and cell lines by real-time quantitative PCR (qRT-PCR) assay while the association of RBM5 expression with prognosis was analyzed by Kaplan-Meier method and compared by log-rank test. Lentivirus was used to overexpress RBM5 in gliomas cells. MTT and BrdU incorporation assay were used to determine cell proliferation and DNA synthesis when the ability of cell migration and invasion was analyzed by transwell assay with/without Matrigel. Cell apoptosis rate was determined with fluorescence-activated cell sorting (FACS) method. Then, expression of apoptosis molecules and critical members in Wnt/β-catenin pathway were detected by western blot analysis.ResultsRBM5 was shown to be downregulated in gliomas tissues and gliomas cell lines. And decreased RBM5 expression was clinically correlated with tumor stage, patient age, and poor prognosis of gliomas patients. The proliferation and DNA synthesis was dramatically inhibited when RBM5 was overexpressed in SHG44 or U251 cells. Also, the ability of cell migration and invasion was disrupted. Then, the level of β-catenin and Cyclin D1 significantly decreased when DKK1 and P-GSK-3β increased reversely in SHG44 cells, which suggested that RBM5 inhibited canonical Wnt/β-catenin signaling. Meanwhile, we demonstrated that caspase3-mediated apoptotic pathway was activated by RBM5 as Bax, TNF-α, and cleaved caspase3 were greatly upregulated while antiapoptotic molecule Bcl-2 was downregulated. Additionally, that apoptotic rate increased significantly from less than 1 to 32% in RBM5-overexpressed SHG44 cells further supported the pro-apoptosis role of RBM5 in gliomas cells.ConclusionsRBM5 plays a suppressor role in human gliomas by inhibiting Wnt/β-catenin signaling and inducing cell apoptosis. This study improves our knowledge about the carcinogenesis and progression of human gliomas, which would greatly contribute to the therapy for gliomas patients.

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Section: Discussionmentioning

confidence: 99%

RBM5 inhibits tumorigenesis of gliomas through inhibition of Wnt/β-catenin signaling and induction of apoptosis

et al. 2017

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“…A549 cells were transiently transfected with recombined plasmid with RBM5 gene (GV287-RBM5) or plasmid with scrambled control sequence using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instruction as described previously [ 19 , 20 , 31 ], with 4 μg DNA for 9 × 10 5 A549 cells in a six-well plate. Five hours after the transfection, the cell culture medium was replaced with 10 % fetal bovine serum.…”

Section: Methodsmentioning

confidence: 99%

“…Total protein from cultured cells was extracted according to the previous study [ 31 ]. Briefly, protein concentration was measured by the Protein Assay Kit (Bio-Rad, CA).…”

Section: Methodsmentioning

confidence: 99%

“…The use of animal was in accordance with animal care guidelines, and the protocol was approved by Jilin University Animal Care Committee. A549 xenografts were established, and RBM5 gene was delivered into xenografts by attenuated Salmonella according to previous studies [ 18 , 31 ]. Briefly, BALB/c athymic nude female mice (nu/nu; 4–5 weeks old) were purchased from the Institute of Zoology, Chinese Academy of Sciences (Beijing, China).…”

Section: Methodsmentioning

confidence: 99%

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Overexpression of RBM5 induces autophagy in human lung adenocarcinoma cells

Wang

et al. 2016

World J Surg Onc

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BackgroundDysfunctions 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.MethodsHuman 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.ResultsOur 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.ConclusionsOur 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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“…is able to regulate apoptosis and cell cycle progression by increasing signal transducer and activator of transcription 5B and bone morphogenetic protein 5 expression levels, and reducing nuclear receptor coactivator 3, Pim-1 proto-oncogene serine/threonine kinase, baculoviral IAP repeat containing 3, BCL-2, EGFR and cyclin dependent kinase 2 expression levels (44)(45)(46)(47). In addition, RBM5 was demonstrated to inhibit cyclin A expression and RB phosphorylation, and thereby regulate cell cycle progression and induce G1 arrest (30).…”

Section: Function Of Rbm5 In Human Cellsmentioning

confidence: 99%

Role of RNA‑binding protein 5 in the diagnosis and chemotherapeutic response of lung cancer (Review)

et al. 2018

Oncol Lett

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Lung cancer remains one of the leading causes of cancer-associated mortality in the world. Lung carcinogenesis is frequently associated with deletions or the loss of heterozygosity at the critical chromosomal region 3p21.3, where RNA-binding protein 5 (RBM5) is localized. RBM5 regulates cell growth, cell cycle progression and apoptosis in cell homeostasis. In the lungs, altered RBM5 protein expression leads to alterations in cell growth and apoptosis, with subsequent lung pathogenesis and varied responses to treatment in patients with lung cancer. Detection of RBM5 expression may be a tumor marker for diagnosis, prediction and treatment response in lung cancer, and may be developed as a potential therapeutic target for drug resistant lung cancer. This review discusses the most recent progress on the role of RBM5 in lung cancer.

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Lentiviral vector-mediated RBM5 overexpression downregulates EGFR expression in human non-small cell lung cancer cells

Cited by 10 publications

References 39 publications

RBM5 inhibits tumorigenesis of gliomas through inhibition of Wnt/β-catenin signaling and induction of apoptosis

RBM5 inhibits tumorigenesis of gliomas through inhibition of Wnt/β-catenin signaling and induction of apoptosis

Overexpression of RBM5 induces autophagy in human lung adenocarcinoma cells

Role of RNA‑binding protein 5 in the diagnosis and chemotherapeutic response of lung cancer (Review)

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