MicroRNA-744-5p Suppresses Tumorigenesis and Metastasis of Osteosarcoma Through the MAPK/ERK Signaling Pathway by Targeting TGFB1

Abstract Background: Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents, and accumulating evidence has revealed that microRNAs (miRNAs) exert a crucial part in the progression of OS. Methods: GSE65071 from the GEO database was analyzed and miR-744-5p was found to be the lowest expressed miRNA. Real-time quantitative PCR (qRT-PCR), Western blotting (WB), colony formation assay, 5-Ethynyl-2-Deoxyuridine (EdU) incorporation assay and Transwell migration and invasion assay were performed to examine the effects of miR-744-5p in vitro, Luciferase-reporter assay was performed to detect the interactions between miR-744-5p and its specific target gene. Subcutaneous tumor-forming animal models and tail vein injection lung metastatic models were conducted in animal experiments to detect the effects of miR-744-5p in vivo. Results: miR-744-5p expression was down-regulated in OS cells and tissues. Higher expression of miR-744-5p was related with better clinical prognosis and lower malignancy degree of OS, including cell proliferation, migration and invasion in vitro and vivo. Transforming growth factor-β1 (TGFB1) was negatively regulated by miR-744-5p and could reverse the effects of miR-744-5p on OS proliferation, migration and invasion. The MAPK/ERK signaling pathway was involved in the miR-744-5p/TGFB1 axis. Conclusions: In general, this study suggests that miR-744-5p is a negative regulator of TGFB1, and suppresses OS progression and metastasis via MAPK/ERK signaling pathway.

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RNA-binding protein IMP3 is a novel regulator of MEK1/ERK signaling pathway in the progression of colorectal Cancer through the stabilization of MEKK1 mRNA

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-01994-8 ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Meng Zhang ◽

Senlin Zhao ◽

Cong Tan ◽

Yanzi Gu ◽

Xuefeng He ◽

...

Keyword(s):

Colorectal Cancer ◽

Signaling Pathway ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Erk Signaling ◽

Luciferase Reporter ◽

Migration And Invasion

Abstract Background MEK1/ERK signaling pathway plays an important role in most tumor progression, including colorectal cancer (CRC), however, MEK1-targeting therapy has little effective in treating CRC patients, indicating there may be a complex mechanism to activate MEK1/ERK signaling pathway except RAS activated mechanism. Methods To investigate the clinical significance of IMP3, we analyzed its expression levels in publicly available dataset and samples from Fudan University Shanghai Cancer Center. The effects of IMP3 on proliferation, migration, and invasion were determined by in vitro and in vivo experiments. To investigate the role of IMP3 in colon carcinogenesis, conditional IMP3 knockout C57BL/6 mice was generated. The IMP3/MEKK1/MEK/ERK signaling axis in CRC was screened and validated by RNA-sequencing, RNA immunoprecipitation, luciferase reporter and western blot assays. Results We find RNA binding protein IMP3 directly bind to MEKK1 mRNA 3′-UTR, which regulates its stability, promote MEKK1 expression and sequentially activates MEK1/ERK signaling. Functionally, IMP3 promote the malignant biological process of CRC cells via MEKK1/MEK1/ERK signaling pathway both in vitro and in vivo, Moreover, IMP3−/− mice show decreased the expression of MEKK1 as well as colorectal tumors compared with wild-type mice after treatment with azoxymethane/dextran sodium sulfate. Clinically, the expression of IMP3 and MEKK1 are positive correlated, and concomitant IMP3 and MEKK1 protein levels negatively correlate with metastasis in CRC patients. In addition, MEK1 inhibitor in combination with shRNA-IMP3 have a synergistic effect both in vitro and in vivo. Conclusion Our study demonstrates that IMP3 regulates MEKK1 in CRC, thus activating the MEK1/ERK signaling in the progression of colorectal cancer, Furthermore, these results provide new insights into potential applications for combining MEK1 inhibitors with other target therapy such as IMP3 in preclinical trials for CRC patients.

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RNA-Binding Protein IMP3 is a Novel Regulator of MEK1/ERK Signaling Pathway in the Progression of Colorectal Cancer through the Stabilization of MEKK1 mRNA

10.21203/rs.3.rs-152463/v1 ◽

2021 ◽

Author(s):

Meng Zhang ◽

Senlin Zhao ◽

Cong Tan ◽

Yanzi Gu ◽

Xuefeng He ◽

...

Keyword(s):

Colorectal Cancer ◽

Signaling Pathway ◽

Rna Binding ◽

Binding Protein ◽

Rna Binding Protein ◽

Erk Signaling ◽

Luciferase Reporter ◽

Migration And Invasion

Abstract Background MEK1/ERK signaling pathway plays an important role in most tumor progression, including colorectal cancer (CRC), however, MEK1-targeting therapy has little effective in treating CRC patients, indicating there may be a complex mechanism to activate MEK1/ERK signaling pathway except RAS activated mechanism. Methods To investigate the clinical significance of IMP3, we analyzed its expression levels in publicly available dataset and samples from Fudan University Shanghai Cancer Center. The effects of IMP3 on proliferation, migration, and invasion were determined by in vitro and in vivo experiments. To investigate the role of IMP3 in colon carcinogenesis, conditional IMP3 knockout C57BL/6 mice was generated. The IMP3/MEKK1/MEK/ERK signaling axis in CRC was screened and validated by RNA-sequencing, RNA immunoprecipitation, luciferase reporter and western blot assays. Results We find RNA binding protein IMP3 directly bind to MEKK1 mRNA 3’-UTR, which regulates its stability, promote MEKK1 expression and sequentially activates MEK1/ERK signaling. Functionally, IMP3 promote the malignant biological process of CRC cells via MEKK1/MEK1/ERK signaling pathway both in vitro and in vivo, Moreover, IMP3−/− mice show decreased the expression of MEKK1 as well as colorectal tumors compared with wild-type mice after treatment with azoxymethane/dextran sodium sulfate. Clinically, the expression of IMP3 and MEKK1 are positive correlated, and concomitant IMP3 and MEKK1 protein levels negatively correlate with metastasis in CRC patients. In addition, MEK1 inhibitor in combination with shRNA-IMP3 have a synergistic effect both in vitro and in vivo. Conclusion Our study demonstrates that IMP3 regulates MEKK1 in CRC, thus activating the MEK1/ERK signaling in the progression of colorectal cancer, Furthermore, these results provide new insights into potential applications for combining MEK1 inhibitors with other target therapy such as IMP3 in preclinical trials for CRC patients.

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Has-miR-875-5p Inhibits Tumorigenesis by Targeting USF2 via TGF-β Signaling Pathway in Gastric Cancer

10.21203/rs.3.rs-885737/v1 ◽

2021 ◽

Author(s):

Shenshuo Gao ◽

Zhikai Zhang ◽

Xubin Wang ◽

Yan Ma ◽

Chensheng Li ◽

...

Keyword(s):

Gastric Cancer ◽

Signaling Pathway ◽

Research Direction ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Gastric Cancer Cells ◽

New Research

Abstract Background: Gastric cancer (GC) is one of the most common malignancies, and more and more evdiences show that the pathogenesis is regulated by various miRNAs.In this study, we investigated the role of miR-875 in GC. Methods:The expression of miR-875-5p was detected in human GC specimens and cell lines by miRNA RT-PCR. The effect of miR-875-5p on GC proliferation was determined by CCK-8 proliferation assay and EDU assay. Migration and invasion were examined by transwell migration and invasion assay and wound healing assay. The interaction between miR-875-5p and its target gene USF2 was verified by a dual luciferase reporter assay. The effects of miR-875-5p in vivo were studied in xenograft nude mice models.Related proteins were detected by Western blot.Results:The results showed that miR-875-5p inhibited the proliferation, migration and invasion of gastric cancer cells in vitro, and inhibited tumorigenesis in vivo. USF2 proved to be a direct target of miR-875-5p. Knockdown of USF2 partially counteracts the effects of miR-875-5p inhibitors.Overexpression of miR-875-5p can inhibit proliferation, migration, and invasion through the TGF-β signaling pathway by down-regulation of USF2 in GC, providing a new research direction for the diagnosis and targeted therapy of GC.Conclusions: MiR-875-5pcan inhibited the progression of GC by directly targeting USF2 and negatively regulating TGF-β signaling pathway.In the future, miR-875-5p is expected to be used as a potential therapeutic target for GC therapy.

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Inhibition of TGF-β Signaling in Gliomas by the Flavonoid Diosmetin Isolated from Dracocephalum peregrinum L.

Molecules ◽

10.3390/molecules25010192 ◽

2020 ◽

Vol 25 (1) ◽

pp. 192 ◽

Cited By ~ 2

Author(s):

Yuli Yan ◽

Xingyu Liu ◽

Jie Gao ◽

Yin Wu ◽

Yuxin Li

Keyword(s):

Signaling Pathway ◽

Caspase 3 ◽

Transforming Growth Factor ◽

Glioma Cells ◽

Chinese Herbs ◽

Migration And Invasion ◽

Western Blots ◽

E Cadherin

Background: Dracocephalum peregrinum L., a traditional Kazakh medicine, has good expectorant, anti-cough, and to some degree, anti-asthmatic effects. Diosmetin (3′,5,7-trihydroxy-4′-methoxyflavone), a natural flavonoid found in traditional Chinese herbs, is the main flavonoid in D. peregrinum L. and has been used in various medicinal products because of its anticancer, antimicrobial, antioxidant, estrogenic, and anti-inflammatory effects. The present study aimed to investigate the effects of diosmetin on the proliferation, invasion, and migration of glioma cells, as well as the possible underlying mechanisms. Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), scratch wound, and Transwell assays were used to demonstrate the effects of diosmetin in glioma. Protein levels of Bcl-2, Bax, cleaved caspase-3, transforming growth factor-β (TGF-β), E-cadherin, and phosphorylated and unphosphorylated smad2 and smad3 were determined by Western blots. U251 glioma cell development and progression were measured in vivo in a mouse model. Results: Diosmetin inhibited U251 cell proliferation, migration, and invasion in vitro, the TGF-β signaling pathway, and Bcl-2 expression. In contrast, there was a significant increase in E-cadherin, Bax, and cleaved caspase-3 expression. Furthermore, it effectively reduced the tumorigenicity of glioma cells and promoted apoptosis in vivo. Conclusion: The results of this study suggest that diosmetin suppresses the growth of glioma cells in vitro and in vivo, possibly by activating E-cadherin expression and inhibiting the TGF-β signaling pathway.

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EPDR1, Which Is Negatively Regulated by miR-429, Suppresses Epithelial Ovarian Cancer Progression via PI3K/AKT Signaling Pathway

Frontiers in Oncology ◽

10.3389/fonc.2021.751567 ◽

2021 ◽

Vol 11 ◽

Author(s):

Zhendan Zhao ◽

Zhiling Wang ◽

Pengling Wang ◽

Shujie Liu ◽

Yingwei Li ◽

...

Keyword(s):

Ovarian Cancer ◽

Epithelial Ovarian Cancer ◽

Signaling Pathway ◽

Cancer Progression ◽

Figo Stage ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Gynecology And Obstetrics

Epithelial ovarian cancer (EOC) is the main pathological type of ovarian cancer. In this study, we found that ependymin-related 1 (EPDR1) was remarkably downregulated in EOC tissues, and low EPDR1 expression was associated with International Federation of Gynecology and Obstetrics (FIGO) stage, metastasis, and poor prognosis. We confirmed that EPDR1 overexpression dramatically suppressed EOC cell proliferation, migration, and invasion in vitro and in vivo. Mechanistically, EPDR1 inhibited EOC tumorigenesis and progression, at least in part, through the repression of the PI3K (Phosphoinositide 3-kinase)/AKT (AKT Serine/Threonine Kinase 1) signaling pathway. Furthermore, the expression and function of EPDR1 were regulated by miR-429, as demonstrated by luciferase reporter assays and rescue experiments. In conclusion, our study validated that EPDR1, negatively regulated by miR-429, played an important role as a tumor-suppressor gene in EOC development via inhibition of the PI3K/AKT pathway. The miR-429/EPDR1 axis might provide novel therapeutic targets for individualized treatment of EOC patients in the future.

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miR-29a-3p inhibits endometrial cancer cell proliferation, migration and invasion by targeting VEGFA/CD C42/PAK1

BMC Cancer ◽

10.1186/s12885-021-08506-z ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Aizhi Geng ◽

Lin Luo ◽

Fengyun Ren ◽

Ling Zhang ◽

Haiying Zhou ◽

...

Keyword(s):

Endometrial Cancer ◽

Signaling Pathway ◽

Scratch Test ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Cancer Cell Proliferation ◽

Ec Cells ◽

Formation Experiment

Abstract Background This study aimed to investigate the mechanism of miR-29a-3p in regulating endometrial cancer (EC) progression. Methods A total of 72 EC patients were enrolled. EC cells were transfected. Cells proliferation, cloning ability, migration and invasion were researched by MTT assay, colony formation experiment, cell scratch test and Transwell experiment respectively. Dual-luciferase reporter assay was performed. Xenograft experiment was conducted using nude mice. miR-29a-3p, VEGFA, CDC42, PAK1 and p-PAK1 expression in cells/tissues was investigated by qRT-PCR and Western blot. Results miR-29a-3p expression was aberrantly reduced in EC patients, which was associated with poor outcome. miR-29a-3p inhibited EC cells proliferation, cloning formation, migration and invasion (P < 0.05 or P < 0.01 or P < 0.001). miR-29a-3p inhibited CDC42/PAK1 signaling pathway activity in EC cells (P < 0.01). VEGFA expression was directly inhibited by miR-29a-3p. miR-29a-3p suppressed EC cells malignant phenotype in vitro and growth in vivo by targeting VEGFA/CDC42/PAK1 signaling pathway (P < 0.05 or P < 0.01). Conclusion miR-29a-3p inhibits EC cells proliferation, migration and invasion by targeting VEGFA/CDC42/PAK1 signaling pathway.

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The oncogenic TBX3 is a downstream target and mediator of the TGF-β1 signaling pathway

Molecular Biology of the Cell ◽

10.1091/mbc.e13-05-0273 ◽

2013 ◽

Vol 24 (22) ◽

pp. 3569-3576 ◽

Cited By ~ 36

Author(s):

Jarod Li ◽

Marc S. Weinberg ◽

Luiz Zerbini ◽

Sharon Prince

Keyword(s):

Signaling Pathway ◽

Transforming Growth Factor ◽

Preliminary Evidence ◽

Transforming Growth Factor Β1 ◽

Migration And Invasion ◽

Mrna Levels ◽

Breast Epithelial ◽

Tgf Β1

The T-box transcription factor, TBX3, plays an important role in embryonic development, and haploinsufficiency of TBX3 causes ulnar–mammary syndrome. Overexpression of TBX3, on the other hand, is associated with several cancers, and preliminary evidence suggests that increased levels of TBX3 may inhibit cell proliferation but promote tumor migration and invasion. Although this suggests that deregulated levels of TBX3 are deleterious in development and promotes disease, very little is known about the signaling pathways that regulate TBX3 expression. Here we show that overexpressing TBX3 inhibits proliferative ability while promoting the migration of breast epithelial cells. We demonstrate that the transforming growth factor β1 (TGF-β1) pathway up-regulates TBX3 protein and mRNA levels and show a detailed transcriptional mechanism by which this occurs. Using in vitro and in vivo assays, we show that Smad3/4 and JunB bind and cooperatively regulate TBX3 promoter activity through a Smad-binding element at −67 base pairs. Further, we show that TBX3 plays a pivotal role in mediating the antiproliferative and promigratory role of TGF-β1 in breast epithelial and skin keratinocytes. This study identifies the TGF-β1 signaling pathway as a potentially important player in the regulation of TBX3 in development and cancer.

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Anthocyanins from Hibiscus syriacus L. Inhibit Melanogenesis by Activating the ERK Signaling Pathway

Biomolecules ◽

10.3390/biom9110645 ◽

2019 ◽

Vol 9 (11) ◽

pp. 645 ◽

Cited By ~ 5

Author(s):

Wisurumuni Karunarathne ◽

Ilandarage Molagoda ◽

Sang Park ◽

Jeong Kim ◽

Oh-Kyu Lee ◽

...

Keyword(s):

Signaling Pathway ◽

Negative Regulator ◽

Erk Signaling ◽

Zebrafish Larvae ◽

B16f10 Cells ◽

Extracellular Signal ◽

Melanin Contents ◽

Hibiscus Syriacus

Hibiscus syriacus L. exhibited promising potential as a new source of food and colorants containing various anthocyanins. However, the function of anthocyanins from H. syriacus L. has not been investigated. In the current study, we evaluated whether anthocyanins from the H. syriacus L. varieties Pulsae and Paektanshim (PS and PTS) inhibit melanin biogenesis. B16F10 cells and zebrafish larvae were exposed to PS and PTS in the presence or absence of α-melanocyte-stimulating hormone (α-MSH), and melanin contents accompanied by its regulating genes and proteins were analyzed. PS and PTS moderately downregulated mushroom tyrosinase activity in vitro, but significantly decreased extracellular and intracellular melanin production in B16F10 cells, and inhibited α-MSH-induced expression of microphthalmia-associated transcription factor (MITF) and tyrosinase. PS and PTS also attenuated pigmentation in α-MSH-stimulated zebrafish larvae. Furthermore, PS and PTS activated the phosphorylation of extracellular signal-regulated kinase (ERK), whereas PD98059, a specific ERK inhibitor, completely reversed PS- and PTS-mediated anti-melanogenic activity in B16F10 cells and zebrafish larvae, which indicates that PS- and PTS-mediated anti-melanogenic activity is due to ERK activation. Moreover, chromatography data showed that PS and PTS possessed 17 identical anthocyanins as a negative regulator of ERK. These findings suggested that anthocyanins from PS and PTS inhibited melanogenesis in vitro and in vivo by activating the ERK signaling pathway.

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ATF5 and HIF1α cooperatively activate HIF1 signaling pathway in esophageal cancer

Cell Communication and Signaling ◽

10.1186/s12964-021-00734-x ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Feng He ◽

Hang Xiao ◽

Yixin Cai ◽

Ni Zhang

Keyword(s):

Esophageal Cancer ◽

Signaling Pathway ◽

Critical Role ◽

Hypoxia Inducible Factor ◽

Expression Level ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Cell Viability Assay

Abstract Background Esophageal cancer (ESCA) is one of the most common cancers worldwide and has a very poor prognosis. Hypoxia-inducible factor 1 (HIF1) signaling pathway plays a critical role in tumorigenesis and is therefore considered a potential therapeutic target in the treatment of many cancers. Activating transcription factor 5 (ATF5) facilitates the expression of various genes and has been extensively studied for its potential role in cancer treatment. Methods The expression level of ATF5 in clinic sample was detected by quantitative real time PCR and immunohistochemistry. ATF5 biological function was investigated by western blot, cell cycle analysis, cell viability assay, luciferase reporter assays, colony formation assay, transwell assay, wound healing assay, tube formation assay, and ELISA assay. CHIP and Re-CHIP assay, GST-pulldown, and RNA-sequencing were used to study the cross-talks between ATF5 and HIF1 complex. Mouse xenograft study was utilized to study the correlation of ATF5 and tumor growth in vivo. Student’s t-test or Chi-square test was used for statistical analysis. Results Here, we first found ATF5 was dramatically upregulated in ESCA cancer and related with poor survival time. Next, we found that the expression level of ATF5 had a positive relationship with the proliferation, migration, and invasion ability of ESCA cells. Besides, we innovatively found that ATF5 functions as a novel coactivator in HIF1 transcription complex by binding to HIF1α. Further, we demonstrated that silencing ATF5 phenocopies HIF1α knockdown in tumorigenic properties in vitro and inhibited ESCA tumor angiogenesis and proliferation in vivo. Conclusion Herein, we found ATF5 as a novel component of the HIF1 transcription complex. The findings of the present study may provide new insights into the development of a novel and more efficient therapeutic strategy against ESCA.

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Has-miR-875-5p Inhibits Tumorigenesis By Targeting USF2 Via TGF-β Signaling Pathway in Gastric Cancer

10.21203/rs.3.rs-885737/v2 ◽

2021 ◽

Author(s):

Shenshuo Gao ◽

Zhikai Zhang ◽

Xubin Wang ◽

Yan Ma ◽

Chensheng Li ◽

...

Keyword(s):

Gastric Cancer ◽

Signaling Pathway ◽

Research Direction ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Gastric Cancer Cells ◽

New Research ◽

Related Proteins

Abstract Background: Gastric cancer (GC) is one of the most common malignancies, and more and more evdiences show that the pathogenesis is regulated by various miRNAs. In this study, we investigated the role of miR-875 in GC. Methods: The expression of miR-875-5p was detected in human GC specimens and cell lines by miRNA RT-PCR. The effect of miR-875-5p on GC proliferation was determined by CCK-8 proliferation assay and EDU assay. Migration and invasion were examined by transwell migration and invasion assay and wound healing assay. The interaction between miR-875-5p and its target gene USF2 was verified by a dual luciferase reporter assay. The effects of miR-875-5p in vivo were studied in xenograft nude mice models. Related proteins were detected by Western blot. Results: The results showed that miR-875-5p inhibited the proliferation, migration and invasion of gastric cancer cells in vitro, and inhibited tumorigenesis in vivo. USF2 proved to be a direct target of miR-875-5p. Knockdown of USF2 partially counteracts the effects of miR-875-5p inhibitors. Overexpression of miR-875-5p can inhibit proliferation, migration, and invasion through the TGF-β signaling pathway by down-regulation of USF2 in GC, providing a new research direction for the diagnosis and targeted therapy of GC.Conclusions: MiR-875-5p can inhibited the progression of GC by directly targeting USF2 and negatively regulating TGF-β signaling pathway. In the future, miR-875-5p is expected to be used as a potential therapeutic target for GC therapy.

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