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Effect of microRNA-128 on cisplatin resistance of glioma SHG-44 cells by targeting JAG1
Dong-Ye Yi1, Qing Su1, Fang-Cheng Zhang1
1Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China.
Abstract:
This current study intends to investigate the effect of microRNA-128 (miR-128) on cisplatin (DDP) resistance in glioma SHG-44 cells. SHG-44/DDP cells were transfected with miR-128 antisense oligonucleotide (ASO) and assigned into blank, resistance, NC, anti-miR-128, miR-128 mimic, si-JAG1, and anti-miR-128 + si-JAG1 groups. qRT-PCR and Western blotting were employed for determining expression of miR-128, JAG1, Bax and Bcl-2. MTT assay, Giemsa staining, and flow cytometry were applied to detect DDP resistance, cellular morphology, and cell cycle, respectively. JAG1 is targeted and negatively regulated by miR-128. In in vitro experiments, compared with the blank group, the rest groups exhibited declined miR-28 and Bax expression, lowered cell inhibition rate and apoptosis rate, but elevated JAG1 and Bcl-2 expression with cells arrested in the S phase. Compared with the resistance group, the anti-miR-128 group showed decreasedBax expression along with a lowered cell inhibition rate and apoptosis rate, but increased JAG1 and Bcl-2 expression with reduced cells arrested in the S phase; while the miR-128 mimic group showed an opposite trend; the si-JAG1 group showed decreased Bcl-2 expression and reduced cells in the S phase. In in vivo experiments, compared with the resistance group, the tumor growth rate, tumor volume, and weight as well as JAG1 expression accelerated in the anti-miR-128 group; whereas the miR-128 mimic and si-JAG1 groups exhibited an opposite trend. Our findings demonstrated that miR-128 ASO transfection might down-regulate the expression of miR-128 in SHG-44/DDP and up-regulate the DDP resistance in SHG-44/DDP cells, providing a potential treatment target for glioma.
Insights
MicroRNA-128 (miR-128) downregulation increases cisplatin resistance in glioma cells by upregulating JAG1. Inhibiting miR-128 offers a potential therapeutic strategy for glioma treatment.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Glioblastoma multiforme (GBM) is an aggressive brain tumor with limited treatment options.
- Cisplatin (DDP) is a chemotherapy agent used in glioma treatment, but resistance is a major challenge.
- MicroRNAs (miRNAs) play crucial roles in gene regulation and cancer progression, including chemoresistance.
Purpose of the Study:
- To investigate the role of microRNA-128 (miR-128) in cisplatin (DDP) resistance in glioma SHG-44 cells.
- To explore the regulatory relationship between miR-128 and JAG1 in the context of DDP resistance.
- To evaluate the potential of targeting miR-128 as a therapeutic strategy for overcoming DDP resistance in glioma.
Main Methods:
- Cell culture of glioma SHG-44 and SHG-44/DDP (cisplatin-resistant) cells.
- Transfection with miR-128 antisense oligonucleotide (ASO), miR-128 mimic, and si-JAG1.
- Quantitative real-time PCR (qRT-PCR) and Western blotting to assess gene and protein expression (miR-128, JAG1, Bax, Bcl-2).
- MTT assay for cell viability, Giemsa staining for cell morphology, and flow cytometry for cell cycle analysis.
- In vivo studies using tumor xenograft models to evaluate tumor growth, volume, and weight.
Main Results:
- Downregulation of miR-128 (using ASO) in SHG-44/DDP cells led to increased DDP resistance, elevated JAG1 and Bcl-2 expression, decreased Bax expression, and S-phase cell cycle arrest.
- Overexpression of miR-128 (using mimic) reversed these effects, reducing DDP resistance and altering the expression of JAG1, Bax, and Bcl-2.
- Silencing JAG1 (using si-JAG1) decreased Bcl-2 expression and reduced S-phase arrest, partially mimicking the effects of miR-128 overexpression.
- In vivo experiments confirmed that anti-miR-128 treatment promoted tumor growth, while miR-128 mimic and si-JAG1 treatments inhibited it.
Conclusions:
- MicroRNA-128 directly targets and negatively regulates JAG1.
- Downregulation of miR-128 contributes to cisplatin resistance in glioma cells by upregulating JAG1 and altering apoptosis-related protein expression (Bax, Bcl-2).
- Targeting miR-128 or JAG1 presents a promising therapeutic avenue for overcoming cisplatin resistance in glioma.
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