scholarly journals Molecular mechanism of microRNA-26a regulation of phosphatase and tensin homolog gene in condyloma acuminatum and penile squamous cell carcinoma

2021 ◽  
Vol 49 (7) ◽  
pp. 030006052110143
Author(s):  
Yayu Hu ◽  
Enping Hu ◽  
Xiangchuan Su ◽  
Xiangen Chen ◽  
Xiulin Tao ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Target Gene ◽  
Bioinformatics Analysis ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Phosphatase And Tensin Homolog ◽  
Tensin Homolog ◽  
Pten Mrna ◽  
Penile Squamous Cell Carcinoma

Objective To investigate the expression levels and mechanisms of microRNA (miRNA) 26a (miR-26a) and phosphatase and tensin homolog (PTEN) in patients with human papillomavirus (HPV)-induced condyloma acuminatum (CA) and penile squamous cell carcinoma (PSCC). Methods Thirty-one patients with HPV-positive CA and 28 with HPV-positive PSCC were included in this retrospective, cross-sectional study. PTEN mRNA and miR-26a levels in lesion tissues, blood, and urine were analyzed by quantitative reverse transcription polymerase chain reaction, and PTEN protein was detected by western blot and enzyme-linked immunosorbent assay. Cell proliferation was assessed by MTT assay. The interaction between miR-26a and PTEN was predicted by bioinformatics analysis and confirmed by dual luciferase reporter assay Results PTEN mRNA and protein levels were significantly lower and miR-26a levels were significantly higher in all samples from patients with PSCC compared with the CA group. Bioinformatics analysis and luciferase reporter assay confirmed PTEN as a target gene of miR-26a. Up-regulation of miR-26a significantly increased the proliferation of Penl1 PSCC cells. Conclusions PTEN expression is down-regulated and miR-26a levels are up-regulated in PSCC compared with CA. PTEN is a direct target gene of miR-26a. These results suggest that miR-26a might regulate HPV-positive progression from CA to PSCC through modulating PTEN.

scholarly journals miR-617 represses SERPINE1 production and reduces tumorigenic traits in oral squamous cell carcinoma cells

2021 ◽  
Author(s):  
Chunguang Zhao ◽  
Zhiyun Liu
Keyword(s):  
Squamous Cell Carcinoma ◽  
Oral Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Lines ◽  
Squamous Cell ◽  
Cell Apoptosis ◽  
Bioinformatics Analysis ◽  
Luciferase Reporter Assay ◽  
Luciferase Reporter ◽  
Reporter Assay

Background: Serpin family E member 1 (SERPINE1) is a serine proteinase inhibitor (serpin) upregulated in diverse types of cancer, including Oral squamous cell carcinoma (OSCC) and functions as an oncogenic role. Hence, exploring pathological mechanism underlying SERPINE1 high expression is crucial to the targeted therapy of OSCC. Methods: Bioinformatics analysis was performed to identify the miRNA and the candidate gene contributing to OSCC progression. The viability, proliferation and apoptosis of the OSCC cell were evaluated using CCK-8 assay, BrdU assay, and cell apoptosis assay, respectively. The RNA pull-down assay and luciferase reporter assay were conducted to verify the relationship between SERPINE1 and miR-617. Results: SERPINE1 was aberrantly upregulated in OSCC tissues and cell lines. Genetically inhibiting SERPINE1 led to reduction of OSCC cell viability and proliferation while elevation of OSCC cell apoptosis. By bioinformatics analysis, miR-617 contained a response element for SERPINE1 overexpression, which is validated by the RNA pull-down and luciferase reporter assay. Furthermore, miR-617 was detected to be downregulated in OSCC tissues and cell lines as well as displayed a negative correlation with advanced stage. Besides, miR-617 mimic or inhibitor transfection could suppress or boost the SERPINE1 expression. More importantly, miR-617 mimic could block the effect of SERPINE1 overexpression on OSCC cells proliferation, viability and apoptosis. Conclusion: SERPINE1 acted as a pro-proliferative oncogenic factor which is partly regulated by miR-167 downregualtion in OSCC cells. Therefore, miR-617/SERPINE1 axis is a potential therapeutic target against OSCC.

scholarly journals MicroRNA-381 Negatively Regulates TLR4 Signaling in A549 Cells in Response to LPS Stimulation

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Zhihao Xu ◽  
Dapeng Dong ◽  
Xiaofei Chen ◽  
Huaqiong Huang ◽  
Shenglan Wen
Keyword(s):  
Target Gene ◽  
Target Genes ◽  
Bioinformatics Analysis ◽  
Respiratory Infections ◽  
A549 Cells ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Elisa Kit ◽  
Cox 2

It is widely reported that miR-381 is dysregulated in various tumors. However, the specific role of miR-381 in respiratory infections has not been reported. To probe this role, A549 cells were pretreated with 1 μg/mL LPS for 24 h. The level of miR-381 was detected using RT-qPCR. The expression of proinflammatory cytokines was determined using an ELISA kit and western blotting. Bioinformatics analysis was used to predict the target genes of miR-381, and a luciferase reporter assay was used to validate the expression of the target genes. miR-381 expression was increased in A549 cells treated with LPS, which is a ligand of TLRs. Further study revealed that the overexpression of miR-381 increased the activity of NF-κB signaling, thereby increasing the expression of IL-6, TNFα, and COX-2. Further study revealed that IκBα was a target gene of miR-381. The upregulation of miR-381 under LPS stimulation contributes to respiratory infections mainly by targeting IκBα.

scholarly journals miR-145 sensitizes esophageal squamous cell carcinoma to cisplatin through directly inhibiting PI3K/AKT signaling pathway

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Tian-Liang Zheng ◽  
De-Ping Li ◽  
Zhan-Feng He ◽  
Song Zhao
Keyword(s):  
Cell Cycle ◽  
Squamous Cell Carcinoma ◽  
Esophageal Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Akt Signaling ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Akt Signaling Pathway ◽  
Cycle Arrest ◽  
Apoptotic Protein

Abstract Background Esophageal squamous cell carcinoma (ESCC) is the eighth most common cancer worldwide and is one of the most lethal malignancies. Cisplatin (DDP) is a key drug for ESCC treatment, but the presence of chemotherapy resistance limits the use of DDP. To enhance chemosensitivity to DDP is important for ESCC treatment. Methods qRT-PCR and Western blotting detected mRNA and protein expression in ESCC tissues and cells. Luciferase reporter assay assessed the interaction between miR-145 and AKT3. Cell cycle, apoptosis and proliferation were investigated with flow cytometry and MTT assay, respectively. Nude mice xenograft model was established, and immunohistochemistry (IHC) and TUNEL assay were conducted to detect Ki-67 level and apoptosis in xenograft tumor. Results Down-regulated miR-145 and up-regulated AKT3 were observed in ESCC tissues and cells. Luciferase reporter assay revealed that miR-145 negatively regulated AKT3 through binding to its 3′-UTR. Overexpression of miR-145 or knockdown of AKT3 promoted DDP-induced cell cycle arrest and apoptosis, as well as reduced IC50 of DDP treatment, which was reversed by AKT3 overexpression. The expression level of MRP1, P-gp, CyclinD1, c-Myc and anti-apoptotic protein Bcl-2 were down-regulated, while pro-apoptotic protein Bax was up-regulated by miR-145. Furthermore, overexpression of miR-145 enhanced the DDP-induced tumor growth suppression in vivo. Conclusion miR-145 increased the sensitivity of ESCC to DDP, and facilitated DDP-induced apoptosis, cycle arrest by directly inhibiting PI3K/AKT signaling pathway to decrease multidrug resistance-associated proteins MRP1 and P-gp expression. Improving the efficacy of DDP by boosting the miR-145 level provides a new strategy for treatment of ESCC.

Targeting exosomal miR-488 inhibits head and neck squamous cell carcinoma growth by mediating RAB25

2020 ◽  
Author(s):  
Xueping Wang ◽  
Xiaoyuan Zhu ◽  
Yulin Zhao
Keyword(s):  
Squamous Cell Carcinoma ◽  
Tumor Growth ◽  
Cell Viability ◽  
Cell Carcinoma ◽  
Colony Formation ◽  
Drug Response ◽  
Luciferase Reporter ◽  
Reporter Assay

Abstract Background: Cancer cell-derived exosomes and its packaged miRNAs have been identified to regulate tumor growth and progression. However, its role in head and neck squamous cell carcinoma (HNSCC) and the potential mechanism still need to be further investigated. Methods: RNA sequencing was conducted to select the dysregulated genes in HNSCC. Gene ontology (GO) analysis and TCGA database were performed to analyze the potential candidate genes for HNSCC progression. Cell viability was analyzed using MTT, and colony formation was visualized using crystal violet staining. Luciferase reporter assay was employed to identify the interaction between miR-488 and RAB25. The role of miR-488 and RAB25 in tumor growth and drug response were investigated in vivo and in vitro. Results: The dysregulated genes in HNSCC captured the signaling of exosomes biogenesis dysfunction. Compared with the normal cells NP69, HNSCC cells had enriched exosomes and its packaged miRNAs, including miR-488. Luciferase reporter assay showed that RAB25 is a downstream target of miR-488. RAB25 was downregulated in HNSCC patients and predicted a poor prognosis. MiR-488/RAB25 signaling controlled cancer cell viability and colony formation ability in vitro and growth in vivo. Importantly, targeting miR-488 significantly inhibited tumor growth and promoted drug response to chemotherapy, suggesting a potential therapeutic promising for HNSCC. Conclusion These findings demonstrate a tumor-cell derived exosomal miR-488 promotes tumor growth by targeting RAB25 that could be targeted for HNSCC treatment.

Ginsenoside 20(S)-Rg3 suppresses cell viability in esophageal squamous cell carcinoma via modulating miR-324-5p-targeted PSME3

2021 ◽  
pp. 096032712110173
Author(s):  
Min Jiang ◽  
Yinxing Zhu ◽  
Hong Yu
Keyword(s):  
Squamous Cell Carcinoma ◽  
Esophageal Squamous Cell Carcinoma ◽  
Cell Viability ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Target Gene ◽  
Luciferase Reporter ◽  
Flow Cytometry Analysis ◽  
Proteasome Activator ◽  
Reporter Assays

Ginsenoside 20(S)-Rg3 is identified as an active saponin monomer which derived from red ginseng and is demonstrated to play an anti-tumor role in diverse cancers. MicroRNAs (miRNAs) are important regulators in the progression of cancers, containing esophageal squamous cell carcinoma (ESCC). It was reported that microRNA 324-5p (miR-324-5p) exerted critical functions in some cancers; however, the detailed molecular mechanism of miR-324-5p mediated by 20(S)-Rg3 to suppress cell viability in ESCC has not been explored. Herein, we explored the function of 20(S)-Rg3 or miR-324-5p on ESCC cell viability by MTT assay, colony formation assay, flow cytometry analysis and western blot analysis. The binding of miR-324-5p to its target gene, proteasome activator subunit 3 (PSME3), was confirmed through RNA pull down and luciferase reporter assays. The results indicated that 20(S)-Rg3 significantly inhibited cell viability and the cell cycle and facilitated cell apoptosis. Furthermore, this effect was strengthened with the increased concentration of 20(S)-Rg3. Moreover, we found that miR-324-5p level was increased under 20(S)-Rg3 treatment. Additionally, overexpressed miR-324-5p inhibited ESCC cell viability, and downregulated miR-324-5p recovered inhibited cell viability caused by 20(S)-Rg3. Further exploration verified that miR-324-5p targeted PSME3, and PSME3 deficiency countervailed the effect of miR-324-5p inhibition on ESCC cell viability under 20(S)-Rg3 treatment. Conclusively, 20(S)-Rg3 suppresses cell viability in ESCC via mediating miR-324-5p-targeted PSME3.

scholarly journals Expression of MicroRNA-301a and its Functional Roles in Malignant Melanoma

2016 ◽  
Vol 40 (1-2) ◽  
pp. 230-244 ◽  
Author(s):  
Lei Cui ◽  
Yuejun Li ◽  
Xiaoxing Lv ◽  
Jinqing Li ◽  
Xiaolin Wang ◽  
...  
Keyword(s):  
Malignant Melanoma ◽  
Western Blot ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Phosphatase And Tensin Homolog ◽  
Functional Roles ◽  
Qrt Pcr ◽  
Tensin Homolog

Background/Aims: Although microRNA-301a has been reported to function as an oncogene in many human cancers, the roles of miR-301a in malignant melanoma (MM) is unclear. The present study aims to investigate the functional roles of miR-301a in MM and its possible molecular mechanisms. Methods: Quantitative real-time PCR (qRT-PCR) assay was performed to detect the expression of miR-301a in MM tissues, and analyze its correlation with metastasis and prognosis of MM patients. In vitro, miR-301a was ectopically expressed using overexpression and knock-down strategies, and the effects of miR-301a expression on growth, apoptosis, migration, invasion and chemosensitivity of MM cells were further investigated. Furthermore, the potential and functional target gene was identified by luciferase reporter, qRT-PCR, Western blot assays. Results: We showed that the expression of miR-301a was significantly upregulated in MM tissues, and upregulation of miR-301a correlated with metastasis and poor prognosis of MM patients. Transfection of miR-301a/inhibitor significantly inhibited growth, colony formation, migration, invasion and enhanced apoptosis and chemosensitivity in MM cells, while transfection of miR-301a/mimic could induce the inverse effects on phenotypes of MM cells. Luciferase reporter, qRT-PCR and Western blot assays showed that phosphatase and tensin homolog (PTEN) was a direct and functional target of miR-301a. It was also observed that the Akt and FAK signaling pathways were involved in miR-301/PTEN-promoting MM progression. Conclusion: Taken together, our study suggests that miR-301a may be used as a potential therapeutic target in the treatment of human MM.

scholarly journals MiR-182 regulates cell proliferation and apoptosis in laryngeal squamous cell carcinoma by targeting the CRR9

2019 ◽  
Vol 39 (10) ◽  
Author(s):  
Yuan Lv ◽  
Dong Ye ◽  
Shijie Qiu ◽  
Jian Zhang ◽  
Zhisen Shen ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Target Genes ◽  
Laryngeal Squamous Cell Carcinoma ◽  
Annexin V ◽  
Luciferase Reporter Assay ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
The Impact

Abstract Background: The effect of miR-182 on the expressions of CRR9 in laryngeal squamous cell carcinoma (LSCC) cells, and the impact on invasion and metastasis of LSCC were investigated in the present paper. Methods: The expressions of miR-182 in LSCC tissue and cell line were detected by RT-qPCR. MTT assay and Annexin V staining were used to detect the effects of miR-182 on tumor cells proliferation. Target gene prediction and screening, and luciferase reporter assay were designed to verify downstream target genes of miR-182. The mRNA and protein expressions of CRR9 were detected by qRT-PCR and Western blot. Finally, the expressions of CRR9 were measured by transfecting cells with miR-182 in mice. Results: Compared with normal tissue and cell, the expressions of miR-182 in tumor tissues and cells were much lower. Over-expressions of miR-182 can increase apoptosis rate. Luciferase reporter assay revealed that CRR9 was a downstream gene of miR-182. Reintroduction of CRR9 abolished miR-182-induced LSCC cell growth inhibition. In animal models, over-expressions of miR-182 can reduce tumor weight and promote apoptosis. Conclusion: miR-182 can inhibit the proliferation of LSCC cells by directly inhibiting the expressions of CRR9, thereby suppressing the occurrences and developments of LSCC.

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