A Novel PCR-Based Approach to Discover miRNA Target Genes

Duan, Shiwei; Wang, Yunliang; Wang, Hongwei; Wang, Shufei; Ji, Li; Dai, Dongjun; Jiang, Danjie; Zhang, Xiaoxi; Wang, Qiang

doi:10.7150/ijms.9343

Cited by 4 publications

(2 citation statements)

References 33 publications

(33 reference statements)

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“…To explore the mechanism by which miR-221-3p protects hippocampal neurons from sevoflurane induced nerve injury, we predicted and discovered that CDKN1B was the target gene of miR-221-3p. MiRNA participates in biological processes in human organism mainly by regulating the expression of its target genes [30,31]. Dong-Mei Wu et al shown that miR-142-3p can partially promote the neuronal cell cycle and inhibit cell apoptosis by inhibiting CDKN1B and TIMP3 expressions [32].…”

Section: Discussionmentioning

confidence: 99%

Study on the ameliorating effect of miR-221-3p on the nerve cells injury induced by sevoflurane

Wang

Tian

et al. 2020

International Journal of Neuroscience

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Purpose: Sevoflurane is a widely used anesthetics, however, it has been reported that sevoflurane has neurotoxic effects. Studies have shown that miR-221-3p can ameliorate neuron damage. This study was to investigate whether miR-221-3p could reduce the neurotoxic effect of sevoflurane on nerve cells. Materials and methods: The rat hippocampal neuron cells were treated with sevoflurane or cultured normally. And we constructed neuron cells that overexpressed or low expression of miR-221-3p in the presence or absence of sevoflurane. The cells were transfected with CDKN1B or siCDKN1B, and co-transfected with miR-221-3p mimic and CDKN1B or miR-221-3p inhibitor and siCDKN1B. Cell viability and apoptosis were detected by CCK-8 and flow cytometer. Target gene of miR-221-3p were predicted by TargetScan and luciferase reporter assay. The expressions of related genes were detected by western blotting and quantitative real-time polymerase chain reaction. Results: Sevoflurane decreased miR-221-3p level and increased CDKN1B level, inhibited cell viability and promoted apoptosis. Overexpress of miR-221-3p decreased CDKN1B level, up-regulated cell viability and inhibited apoptosis, and reversed the effects of sevoflurane on cell viability and apoptosis, while the effects low expression of miR-221-3p was contrary. CDKN1B was the target gene of miR-221-3p, which inhibited cell viability and promoted apoptosis, and reversed the effects of miR-221-3p mimic, whereas siCDKN1B did the opposite effects. Conclusions: Sevoflurane can cause nerve cell injury, and miR-221-3p may promote cell activity and inhibit apoptosis by inhibiting CDKN1B expression, thereby ameliorating cell injury induced by sevoflurane.

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Section: Discussionmentioning

confidence: 99%

Study on the ameliorating effect of miR-221-3p on the nerve cells injury induced by sevoflurane

Wang

Tian

et al. 2020

International Journal of Neuroscience

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“…The miR-377 appears as an important aging regulator by regulating, among others, superoxide dismutase 2 enzyme (SOD-2) 13,14 . SOD is the body's main antioxidant, which catalyzes the conversion of superoxide radicals to hydrogen peroxide.…”

Section: Introductionmentioning

confidence: 99%

Efecto del estrés oxidativo sobre la calcificación vascular a través del microARN-377

García

López

Martínez-Arias

et al. 2017

Rev Osteoporos Metab Miner

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Introduction: Oxidative stress has been implicated in the development and progression of vascular calcification (VC). However, this causal association remains a matter of controversy. Objective: To analyze in an experimental model of chronic renal failure (CRF), the effect of oxidative stress on the development and progression of the VC, assessing the implication of microRNA-377 (miR-377). Material and methods: Two groups of Wistar rats with CRF were studied. Group 1 received normal diet in phosphorus (CRF+NP). Group 2 received a high phosphorus (CRF+HP) diet. A group of Sham rats was included. After 20 weeks, the rats were sacrificed. Results: Serum phosphorus and parathormone did not increase in the CRF+HP group compared to CRF+NP, but fibroblast growth factor 23 (FGF23) levels significantly increased. In the CRF+NP group, aortic calcium content increased three-fold over the Sham group, a 17-fold increase in the CRF+HP group, where the bone mineral density in the proximal tibia decreased significantly. In the CRF+NP group, the expression of miR-377 decreased by 65%, with no additional effect detected of the diet with high phosphorus content. In the CRF+NP group, the protein expression of mitochondrial superoxide dismutase 2 (SOD-2) increased 3-fold, and in the CRF+HP group it increased up to 6-fold. Conclusions: CRF, with or without high phosphorus dietary content, triggered the descent of miR-377. Excess phosphorus increased SOD-2 as a compensatory mechanism to curb oxidative stress and vascular damage. Controlling phosphorus content in the diet when the renal impairment function is compromised will reduce the vascular damage produced due oxidative stress, among other factors.

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miR-186 downregulates protein phosphatase PPM1B in bladder cancer and mediates G1-S phase transition

Yang

Yuan

et al. 2015

Tumor Biol.

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Nuclear factor-κB (NF-κB) is a core regulator in multiple tumorigenic pathways. Its activation is mediated by IκB kinase β (IKKβ). Protein phosphatase PPM1B is reported to dephosphorylate IKKβ, thereby terminating IKKβ-mediated NF-κB activation. However, the role of PPM1B in bladder cancer is unclear. The aim of this study was to determine the expression patterns and molecular mechanisms of PPM1B in bladder cancer. Comparative analyses were conducted in six bladder cancer cell lines, a normal urinary epithelial cell line, and adjacent non-tumorous bladder epithelia. Searches were conducted through publicly available algorithms and The Cancer Genome Atlas. HT-1376 and RT4 cells were transduced to stably overexpress PPM1B and its predicted regulator miR-186. Subsequent in vitro studies included 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), colony formation, anchorage-independent growth ability, luciferase reporter assays, and flow cytometric cell cycle analyses. A xenograft model was established in nude mice to evaluate the effect of PPM1B in bladder tumors in vivo. The results revealed that PPM1B was frequently downregulated in bladder cancer cells at both protein and messenger RNA (mRNA) levels, whereas miR-186 was upregulated. Further analyses showed that miR-186 promoted G1-S transition by targeting PPM1B at its 3'-untranslated region (3'UTR). Conversely, ectopic expression of PPM1B significantly suppressed proliferation and tumorigenicity in bladder cancer cells in vitro and in vivo, thereby neutralizing the oncogenic effect of miR-186. This study has identified PPM1B and miR-186 as potential diagnostic markers in bladder cancer. Promotion of PPM1B and suppression of miR-186 may offer effective therapeutic strategies in the treatment of bladder cancer.

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A Novel PCR-Based Approach to Discover miRNA Target Genes

Cited by 4 publications

References 33 publications

Study on the ameliorating effect of miR-221-3p on the nerve cells injury induced by sevoflurane

Study on the ameliorating effect of miR-221-3p on the nerve cells injury induced by sevoflurane

Efecto del estrés oxidativo sobre la calcificación vascular a través del microARN-377

miR-186 downregulates protein phosphatase PPM1B in bladder cancer and mediates G1-S phase transition

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