Expression of MiR-664-3p in Osteosarcoma and Its Effects on the Proliferation and Apoptosis of Osteosarcoma Cells

2020 ◽  
Author(s):  
Ye LI ◽  
Jie TANG ◽  
Yong HU ◽  
Yonghai PENG ◽  
Junwen WANG
Keyword(s):  
Proteolipid Protein ◽  
Biological Indicator ◽  
Negative Control ◽  
Expression Level ◽  
Osteosarcoma Cells ◽  
Apoptosis Rate ◽  
Relative Expression Level ◽  
Proliferation And Apoptosis ◽  
Polymerase Chain ◽  
Normal Bone Tissue

Background: To explore the expression level of miR-664-3p in osteosarcoma and its effects on the proliferation and apoptosis of osteosarcoma cells. Methods: Specimens of osteosarcoma tissues were collected from 41 cases undergoing surgical treatment in the Orthopedics Department of Wuhan Puai Hospital, Wuhan, China from January 2015 to February 2018. Another 40 cases of normal bone tissue were collected. The expression of miR-664-3p were detected using quantificational real-time polymerase chain reaction. miR-664-3p mimics, miR-664-3p inhibitor and miR-664- 3p negative control (NC) were used to transfect U2-OS, which were named as mimics group, inhibitor group and NC group, respectively. MTT assay was adopted to detect the effects of microRNA-664-3p on the proliferation of U2-OS after 24, 48, 72, 96 and 120 hours of transfection. Flow cytometry was applied to measure the apoptosis rate of U2-OS after miR-664-3p transfection. Finally, Western Blot was employed to detect the expression of proteolipid protein 2 (PLP2). Results: The total apoptosis rate of cells in the inhibitor group was obviously higher than those in the mimics group and the NC group (P<0.001). The relative expression level of PLP2 in the inhibitor group was significantly lower than those in the mimics group and the NC group (P<0.001). Conclusion: MiR-664-3p may be involved in the occurrence and development of osteosarcoma, and can regulate the proliferation and apoptosis of U2-OS cells, and the expression of PLP2. Besides, miR-664-3p may become a novel molecular biological indicator for the diagnosis, targeted treatment and prognosis assessment of osteosarcoma.

Mir-29c Expression in Glioma and Its Effects on Tumor Cell Proliferation and Apoptosis

2020 ◽  
Author(s):  
Peiquan HUI ◽  
Yuling WANG ◽  
Bing CHEN ◽  
Zengwu WANG ◽  
Shiqiang QIN
Keyword(s):  
Cell Proliferation ◽  
Glioma Cells ◽  
Negative Control ◽  
Expression Level ◽  
Control Group ◽  
Tumor Differentiation ◽  
Blank Group ◽  
Pathological Type ◽  
Proliferation And Apoptosis ◽  
Experimental Group

Background: To investigate the expression of microRNA-29c (miR-29c) in glioma and its effects on cell proliferation and apoptosis. Methods: A retrospective analysis was performed on 76 glioma patients in People's Hospital of Weifang, Weifang, Shandong, China from May 2013 to June 2017 (experimental group) and 63 healthy subjects in the same period (control group). qRT-PCR was used to detect the miR-29c expression. Changes of serum miR-29c expression level and the correlation of miR-29c of glioma patients with the degree of tumor differentiation and pathological type were observed. Cells were grouped before transfection into blank group (no transfection), negative control group (transfected with miRNA NC) and experimental group (transfected with miR-29c mimics). CCK-8 assay was used to detect cell proliferation, flow cytometry to detect apoptosis. Results: Expression of miR-29c in serum was significantly lower in experimental group than that in control group (P<0.05). The expression level of miR-29c of glioma patients increased with the degree of tumor differentiation (P<0.05). miR-29c in serum was not significantly correlated with the pathological type. Conclusion: miR-29c could inhibit the proliferation of glioma cells and promote apoptosis. miR-29c is lowly expressed in glioma, and the overexpression of which in glioma cells can inhibit tumor cells proliferation and promote apoptosis. It may be a tumor suppressor miRNA of glioma, and the expression level of which can be used as reference for evaluating the grade of glioma. It is indicated that the abnormal expression of miR-29c may be a key factor in the occurrence and development of glioma.

scholarly journals Expression of Talin-1 in endometriosis and its possible role in pathogenesis

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Xian Tang ◽  
Qing Li ◽  
Lijie Li ◽  
Jianfa Jiang
Keyword(s):  
Cell Invasion ◽  
Expression Level ◽  
Migration And Invasion ◽  
Endometrial Stromal Cells ◽  
Invasion And Migration ◽  
Proliferation And Apoptosis ◽  
Mrna And Protein Expression ◽  
Polymerase Chain ◽  
And Migration

Abstract Background Endometriosis is a disease that involves active cell invasion and migration. Talin-1 can promote cell invasion, migration and adhension in various cancer cells, but its role in endometriosis has not been investigated. This study was to investigate the expression level of Talin-1 in endometriosis and the role of Talin-1 in the proliferation, adhesion, migration, and invasion of human endometrial stromal cells (ESCs). Methods Ectopic and eutopic endometrial tissues were collected from women with endometriosis, and the control endometrial tissues were obtained from patients without endometriosis. The expression level of Talin-1 was detected in each sample using quantitative real-time polymerase chain reaction and immunohistochemistry. The expression of Talin-1 was inhibited using RNA interference in ESCs, and its proliferation, apoptosis, adhesion, migration, and invasion capacity were analyzed. Western blotting was performed to detect the expression of related molecules after the downregulation of Talin-1. Results The results showed that the mRNA and protein expression of Talin-1 were significantly increased in the ectopic endometrium and eutopic endometrial tissues compared with the controls. The knockdown of Talin-1 did not affect the proliferation and apoptosis of ESCs. The results indicated that the downexpression of Talin-1 inhibited the adhesion, invasion, and migration of ESCs. In addition, the expressions of N-cadherin, MMP-2, and integrin β3 were significantly lower after the deregulation of Talin-1, whereas the levels of E-cadherin were significantly increased. Conclusions The expression of Talin-1 was increased in the ectopic and eutopic endometrial tissues compared with the control endometrium. The downregulation of Talin-1 inhibited the adhesion, invasion, and migration of ESCs.

scholarly journals Role of microRNA-25 in high glucose cultured Müller glia

2021 ◽  
Vol 14 (5) ◽  
pp. 643-648
Author(s):  
Yu Liu ◽  
◽  
Song-Tao Yuan ◽  
Qing-Huai Liu ◽  
◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Glucose ◽  
Cultured Cells ◽  
Negative Control ◽  
Expression Level ◽  
Cell Counting ◽  
Muller Glia ◽  
Müller Glia ◽  
Proliferation And Apoptosis

AIM: To investigate the role of microRNA-25 (miR-25) in proliferation and apoptosis of retinal Müller glia (MG) under high glucose condition. METHODS: The purity of the cultured cells was verified by immunocytochemistry and flow cytometry using antibodies that specifically recognized MG. The expression level of miR-25 under normal and high glucose conditions were validated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). miR-25 mimics and negative control were transfected into MG and multiple functional experiments including cell counting kit-8 assay, EDU assay, and flow cytometry were conducted to explore the effects of miR-25 on the proliferation and apoptosis of high glucose cultured MG (HGMG). RESULTS: Immunocytochemistry and flow cytometry confirmed the high purity of primary cultured MG. RT-PCR results showed that the expression level of miR-25 was significantly repressed in HGMG, while over-expression of miR-25 by miR-25 mimic markedly inhibited the high glucose induced cell apoptosis and promoted the proliferation of MG. CONCLUSION: The expression level of miR-25 is significantly downregulated in HGMG and its overexpression could attenuate the high glucose damages on MG by promoting proliferation and reducing apoptosis.

scholarly journals Assessment of the Nutraceutical Effects of Oleuropein and the Cytotoxic Effects of Adriamycin, When Administered Alone and in Combination, in MG-63 Human Osteosarcoma Cells

Nutrients ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 354
Author(s):  
Katerina Gioti ◽  
Anastasia Papachristodoulou ◽  
Dimitra Benaki ◽  
Nektarios Aligiannis ◽  
Alexios-Leandros Skaltsounis ◽  
...  
Keyword(s):  
Chemotherapeutic Agent ◽  
Molecular Techniques ◽  
Osteosarcoma Cells ◽  
Elisa Method ◽  
Chain Reaction ◽  
Cytotoxic Effects ◽  
Human Osteosarcoma ◽  
Human Osteosarcoma Cells ◽  
Anti Cancer ◽  
Polymerase Chain

Oleuropein (OLEU) is the most distinguished phenolic compound found in olive fruit and the leaves of Olea europaea L., with several pharmacological properties, including anti-cancer actions. Adriamycin (ADR) is an anthracycline widely used as a chemotherapeutic agent, although it presents significant side effects. The aim of the present study was to investigate the effect of oleuropein alone (20 μg/mL) and in co-treatment with ADR (50 nM), in MG-63 human osteosarcoma cells. Therefore, cellular and molecular techniques, such as MTT assay, flow cytometry, real-time Polymerase Chain Reaction (PCR), western blot and Elisa method, as well as Nuclear Magnetic Resonance (NMR) spectroscopy, were applied to unveil changes in the signal transduction pathways involved in osteosarcoma cells survival. The observed alterations in gene, protein and metabolite levels denote that OLEU not only inhibits MG-63 cells proliferation and potentiates ADR’s cytotoxicity, but also exerts its action, at least in part, through the induction of autophagy.

scholarly journals Decitabine shows anti-acute myeloid leukemia potential via regulating the miR-212-5p/CCNT2 axis

2020 ◽  
Vol 15 (1) ◽  
pp. 1013-1023
Author(s):  
Lina Xing ◽  
Jinhai Ren ◽  
Xiaonan Guo ◽  
Shukai Qiao ◽  
Tian Tian
Keyword(s):  
Acute Myeloid Leukemia ◽  
Cell Viability ◽  
Cell Apoptosis ◽  
Myeloid Leukemia ◽  
Luciferase Reporter ◽  
Expression Levels ◽  
Proliferation And Apoptosis ◽  
Polymerase Chain ◽  
The Relationship ◽  
Acute Myeloid

AbstractPrevious research has revealed the involvement of microRNA-212-5p (miR-212-5p) and cyclin T2 (CCNT2) in acute myeloid leukemia (AML). However, whether the miR-212-5p/CCNT2 axis is required for the function of decitabine in AML has not been well elucidated. Quantitative reverse transcription-polymerase chain reaction was used to examine enrichment of miR-212-5p. The relationship between CCNT2 and miR-212-5p was verified by the luciferase reporter assay. Cell apoptosis was evaluated by flow cytometry and western blot. CCK-8 assay was performed to determine cell viability. Decitabine significantly repressed cell viability, while promoted cell apoptosis. Meanwhile, the expression levels of cyclinD1, CDK4, and Bcl-2 were suppressed in cells with decitabine exposure, but Bax and caspase-3 expression levels were upregulated. Besides, miR-212-5p upregulation had the similar function with decitabine in AML cell proliferation and apoptosis. Subsequently, restoration of CCNT2 attenuated miR-212-5p overexpression-induced effects in Kasumi-1 and SKNO-1 cells. In addition, miR-212-5p depletion reversed decitabine-induced CCNT2 downregulation. The miR-212-5p/CCNT2 axis had an implication in the anti-leukemic effect of decitabine in AML.

Expression level and differential JAK2-V617F–binding of the adaptor protein Lnk regulates JAK2-mediated signals in myeloproliferative neoplasms

Blood ◽  
2010 ◽  
Vol 116 (26) ◽  
pp. 5961-5971 ◽  
Author(s):  
Fanny Baran-Marszak ◽  
Hajer Magdoud ◽  
Christophe Desterke ◽  
Anabell Alvarado ◽  
Claudine Roger ◽  
...  
Keyword(s):  
Myeloproliferative Neoplasms ◽  
Adaptor Protein ◽  
Jak2 V617f ◽  
Negative Control ◽  
Sh2 Domain ◽  
Expression Level ◽  
Activating Mutations ◽  
Thrombopoietin Receptor ◽  
Dependent Cell

Abstract Activating mutations in signaling molecules, such as JAK2-V617F, have been associated with myeloproliferative neoplasms (MPNs). Mice lacking the inhibitory adaptor protein Lnk display deregulation of thrombopoietin/thrombopoietin receptor signaling pathways and exhibit similar myeloproliferative characteristics to those found in MPN patients, suggesting a role for Lnk in the molecular pathogenesis of these diseases. Here, we showed that LNK levels are up-regulated and correlate with an increase in the JAK2-V617F mutant allele burden in MPN patients. Using megakaryocytic cells, we demonstrated that Lnk expression is regulated by the TPO-signaling pathway, thus indicating an important negative control loop in these cells. Analysis of platelets derived from MPN patients and megakaryocytic cell lines showed that Lnk can interact with JAK2-WT and V617F through its SH2 domain, but also through an unrevealed JAK2-binding site within its N-terminal region. In addition, the presence of the V617F mutation causes a tighter association with Lnk. Finally, we found that the expression level of the Lnk protein can modulate JAK2-V617F–dependent cell proliferation and that its different domains contribute to the inhibition of multilineage and megakaryocytic progenitor cell growth in vitro. Together, our results indicate that changes in Lnk expression and JAK2-V617F–binding regulate JAK2-mediated signals in MPNs.

The Clinical Significance of miR-135b-5p and Its Role in the Proliferation and Apoptosis of Hippocampus Neurons in Children with Temporal Lobe Epilepsy

2020 ◽  
Vol 42 (5-6) ◽  
pp. 187-194
Author(s):  
Ruixiang Li ◽  
Jiahua Hu ◽  
Sue Cao
Keyword(s):  
Temporal Lobe Epilepsy ◽  
Cell Viability ◽  
Temporal Lobe ◽  
Luciferase Reporter ◽  
Diagnostic Biomarker ◽  
Flow Cytometric ◽  
Proliferation And Apoptosis ◽  
Apoptosis Assay ◽  
Polymerase Chain ◽  
Adverse Influence

Temporal lobe epilepsy (TLE) is the most familiar localized epilepsy in children. MicroRNAs (miRNAs) are essential for the inhibition or promotion of numerous diseases. This study aimed to detect the expression of miR-135b-5p and primarily uncover its underlying function and mechanism in children with TLE. Quantitative real-time polymerase chain reaction was used to evaluate the expression of miR-135b-5p in children with TLE and in a rat model of epilepsy. MTT assay and flow cytometric apoptosis assay were conducted to evaluate the effects of miR-135b-5p on cell viability and apoptosis. Additionally, the dual luciferase reporter assay was performed to confirm the direct target of miR-135b-5p. Our data showed that the expression of miR-135b-5p was significantly decreased in children with TLE and in the epileptic rat neuron model. The dysregulation of miR-135b-5p could serve as a promising diagnostic biomarker for children with TLE. The overexpression of miR-135b-5p moderated the adverse influence on cell viability and apoptosis induced by magnesium-free medium. SIRT1 was identified as a target gene of miR-135b-5p. These results proved that miR-135b-5p might serve as a potential diagnostic biomarker in children with TLE. Overexpression of miR-135b-5p alleviates the postepileptic influence on cell viability and apoptosis by targeting SIRT1.

scholarly journals Genotypic diversity of S. mutans in dental biofilm formed in situ under sugar stress exposure

2007 ◽  
Vol 18 (3) ◽  
pp. 185-191 ◽  
Author(s):  
Rodrigo Alex Arthur ◽  
Cínthia Pereira Machado Tabchoury ◽  
Renata de Oliveira Mattos-Graner ◽  
Altair A. Del Bel Cury ◽  
Adriana Franco Paes Leme ◽  
...  
Keyword(s):  
Genotypic Diversity ◽  
Negative Control ◽  
Stress Exposure ◽  
Chain Reaction ◽  
Dental Biofilm ◽  
Polymerase Chain ◽  
Fermentable Carbohydrates ◽  
Almost All ◽  
Frequent Exposure

In situ dental biofilm composition under sugar exposure is well known, but sugar effect on the genotypic diversity of S. mutans in dental biofilm has not been explored. This study evaluated S. mutans genotypic diversity in dental biofilm formed in situ under frequent exposure to sucrose and its monosaccharide constituents (glucose and fructose). Saliva of 7 volunteers was collected for isolation of S. mutans and the same volunteers wore intraoral palatal appliances, containing enamel slabs, which were submitted to the following treatments: distilled and deionized water (negative control), 10% glucose + 10% fructose (fermentable carbohydrates) solution or 20% sucrose (fermentable and EPS inductor) solution, 8x/day. After 3, 7 and 14 days, the biofilms were colleted and S. mutans colonies were isolated. Arbitrarily primed polymerase chain reaction (AP-PCR) of S. mutans showed that salivary genotypes were also detected in almost all biofilm samples, independently of the treatment, and seemed to reflect those genotypes present at higher proportion in biofilms. In addition to the salivary genotypes, others were found in biofilms but in lower proportions and were distinct among treatment. The data suggest that the in situ model seems to be useful to evaluate genotypic diversity of S. mutans, but, under the tested conditions, it was not possible to clearly show that specific genotypes were selected in the biofilm due to the stress induced by sucrose metabolism or simple fermentation of its monosaccharides.

scholarly journals Role of MiR-155 Signal Pathway in Regulating Podocyte Injury Induced by TGF-β1

2017 ◽  
Vol 42 (4) ◽  
pp. 1469-1480 ◽  
Author(s):  
Xu Lin ◽  
Xintng Zhen ◽  
Haiting Huang ◽  
Haohao Wu ◽  
Yanwu You ◽  
...  
Keyword(s):  
Protein Expression ◽  
Antisense Oligonucleotides ◽  
Negative Control ◽  
Signal Pathway ◽  
Caspase 9 ◽  
Podocyte Injury ◽  
Apoptosis Rate ◽  
Mrna And Protein Expression ◽  
Tgf Β1

Background/Aims: Transforming growth factor beta 1 (TGF-β1) plays a critical role in the pathogenesis of glomerulosclerosis. The purpose of this study was to examine the effects of inhibition of miR-155 on podocyte injury induced by TGF-β1 and to determine further molecular mediators involved in the effects of miR-155. Methods: Conditionally immortalized podocytes were cultured in vitro and they were divided into four groups: control; TGF-β1 treatment; TGF-β1 with miR-155 knockdown [using antisense oligonucleotides against miR-155 (ASO-miR-155)] and TGF-β1 with negative control antisense oligonucleotides (ASO-NC). Real time RT-PCR and Western blot analysis were employed to determine the mRNA and protein expression of nephrin, desmin and caspase-9, respectively. Flow cytometry was used to examine the apoptotic rate of podocytes and DAPI fluorescent staining was used to determine apoptotic morphology. In addition, we examined the levels of miR-155, TGF-β1, nephrin, desmin and caspase-9 in glomerular tissues of nephropathy induced by intravenous injections of adriamycin in rats. Results: mRNA and protein expression of desmin and caspase-9 was increased in cultured TGF-β1-treated podocytes, whereas nephrin was decreased as compared with the control group. Importantly, miR-155 knockdown significantly attenuated upregulation of desmin and caspase-9, and alleviated impairment of nephrin induced by TGF-β1. Moreover, the number of apoptotic podocytes was increased after exposure to TGF-β1 and this was alleviated after miR-155 knockdown. Knocking down miR-155 also decreased an apoptosis rate of TGF-β1-treated podocytes. Note that negative control antisense oligonucleotides failed to alter an increase of the apoptosis rate in TGF-β1-treated podocytes. Consistent with in vitro results, expression of miR-155, TGF-β1, desmin and caspase-9 was increased and nephrin was decreased in glomerular tissues with nephropathy in vivo experiments. Conclusions: TGF-β1 impairs the protein expression of nephrin and amplifies the protein expression of desmin and caspase -9 via miR-155 signal pathway. Inhibition of miR-155 alleviates these changes in podocytes-treated with TGF-β1 and attenuated apoptosis of podocytes. Our data suggest that miR-155 plays a role in mediating TGF-β1-induced podocyte injury via nephrin, desmin and caspase-9. Results of the current study also indicate that blocking miR-155 signal has a protective effect on podocyte injury. Targeting one or more of these signaling molecules may present new opportunities for treatment and management of podocyte injury observed in glomerulosclerosis.

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