Effects of FHIT gene on proliferation and apoptosis of osteosarcoma cells

AbstractTen-eleven translocation (TET) proteins are abnormally expressed in various cancers. Osteosarcoma cells were treated with hydroxyurea to investigate the expression pattern of TET proteins in these cells. The expression of TET1 was increased in U2OS cells after treatment with hydroxyurea. In addition, hydroxyurea increased cell apoptosis and altered the cell cycle. TET proteins catalyze the oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC); therefore, 5mC and 5hmC levels were evaluated. Increased 5hmC levels were observed after the hydroxyurea treatment. Experiments examining cell apoptosis and the cell cycle after knockdown and overexpression of TET1 were conducted to further investigate whether TET1 expression affected cell growth. The overexpression of TET1 increased cell apoptosis and inhibited cell growth. Taken together, TET1 expression regulated proliferation and apoptosis in U2OS cells, changes that were associated with 5hmC levels.

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miR-124 Regulates Caveolin-1 Expression and Affects Proliferation and Apoptosis of Osteosarcoma Cells

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2019.2133 ◽

2019 ◽

Vol 9 (9) ◽

pp. 1245-1249

Author(s):

Huanzhi Ma ◽

Jian Wang ◽

Jun Shi ◽

Wei Zhang ◽

Dongsheng Zhou

Keyword(s):

Cell Proliferation ◽

Cell Apoptosis ◽

Luciferase Activity ◽

Cell Number ◽

Osteosarcoma Cell ◽

Osteosarcoma Cells ◽

Specific Mechanism ◽

Caveolin 1 ◽

Proliferation And Apoptosis ◽

Relative Luciferase Activity

Osteosarcoma (OS) seriously affects human health. miR-124 expression is closely related to osteosarcoma, but its specific mechanism remains unclear. Our study intends to evaluate miR-124’s effect on osteosarcoma. MG-63 cells were transfected with miR-124 mimics/NC followed by analysis of miR-124 expression by real-time PCR, cell proliferation by CCK8 assay, cell apoptosis by flow cytometry as well as the level of caveolin-1 (CAV1) by Western blot. miR-124 was significantly lower and CAV1 was increased in the four osteosarcoma cells than those in normal osteoblasts (P < 0.05). miR-124 mimics transfection significantly reduced CAV1 level and cell number (P < 0.05) and increased cell apoptosis rate (P < 0.05). Moreover, miR-124 inhibitor significantly promoted the relative luciferase activity in pmirGLO-CAV1-3′UTR-wt-transfected cells (P < 0.05). miR-124 affects osteosarcoma cell proliferation and apoptosis via targeting CAV1.

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Influence mechanism of miRNA‑144 on proliferation and apoptosis of osteosarcoma cells

Oncology Letters ◽

10.3892/ol.2019.11197 ◽

2019 ◽

Author(s):

Xu Zhang ◽

Zhengwei Li ◽

Wei Ji ◽

Xilong Chen ◽

Qiang Gao ◽

...

Keyword(s):

Osteosarcoma Cells ◽

Influence Mechanism ◽

Proliferation And Apoptosis

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Serine/Threonine Kinase 35, a Target Gene of STAT3, Regulates the Proliferation and Apoptosis of Osteosarcoma Cells

Cellular Physiology and Biochemistry ◽

10.1159/000487172 ◽

2018 ◽

Vol 45 (2) ◽

pp. 808-818 ◽

Cited By ~ 8

Author(s):

Zhong Wu ◽

Jie Liu ◽

Siyuan Hu ◽

Yuchang Zhu ◽

Shaohua Li

Keyword(s):

Mrna Expression ◽

Real Time ◽

Real Time Pcr ◽

Gene Set Enrichment Analysis ◽

Cell Counting ◽

Luciferase Reporter ◽

Osteosarcoma Cells ◽

Serine Threonine Kinase ◽

Threonine Kinase ◽

Proliferation And Apoptosis

Background/Aims: Serine/threonine kinase 35 (STK35) may be associated with Parkinson disease and human colorectal cancer, but there have been no reports on the expression levels or roles of STK35 in osteosarcoma. Methods: STK35 mRNA expression was determined in osteosarcoma and bone cyst tissues by real-time PCR. Cell proliferation and apoptosis were assessed by Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis, respectively. Results: STK35 was up-regulated in osteosarcoma tissues as indicated by analyzing publicly available expression data (GEO dataset E-MEXP-3628) and real-time PCR analysis on our own cohort. We subsequently investigated the effects of STK35 knockdown on two osteosarcoma cell lines, MG63 and U2OS. STK35 knockdown inhibited the growth of osteosarcoma cells in vitro and in xenograft tumors. Meanwhile, STK35 knockdown enhanced apoptosis. Expression of the active forms and the activity of two major executioner caspases, caspase 3 and caspase 7, were also increased in osteosarcoma cells with STK35 silenced. Additionally, Gene Set Enrichment Analysis (GSEA) identified that the JAK/STAT signaling pathway was positively correlated with STK35 expression. The mRNA expression of STK35 was repressed by STAT3 small interfering RNA (siRNA), but not by siRNA of STAT4, STAT5A or STAT6. A luciferase reporter assay further demonstrated that STAT3 transcriptionally regulated STK35 expression. A chromatin immunoprecipitation (ChIP) assay confirmed the direct recruitment of STAT3 to the STK35 promoter. The promotion effects of STAT3 knockdown on cell apoptosis were partially abolished by STK35 overexpression. Furthermore, STK35 mRNA expression was positively correlated with STAT3 mRNA expression in osteosarcoma tissues by Pearson correlation analysis. Conclusions: These results collectively reveal that STAT3 regulates the transcription of STK35 in osteosarcoma. STK35 may exert an oncogenic role in osteosarcoma.

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

Iranian Journal of Public Health ◽

10.18502/ijph.v48i10.3489 ◽

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.

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Effect of microRNA-21 on Proliferation and Apoptosis of Osteosarcoma Cells via Phosphatase and Tensin Homologue (PTEN)-Phosphoinositide 3-Kinase (PI3K)/AKT Pathway

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2020.2293 ◽

2020 ◽

Vol 10 (4) ◽

pp. 512-517

Author(s):

Lei Huang ◽

Yongheng Xie ◽

Zilong Yao ◽

Bin Yu

Keyword(s):

Cell Proliferation ◽

Cell Apoptosis ◽

Akt Signaling ◽

Luciferase Reporter ◽

Osteosarcoma Cells ◽

Luciferase Reporter Gene ◽

Gene Assay ◽

Proliferation And Apoptosis ◽

Phosphoinositide 3 Kinase ◽

Phosphatase And Tensin Homologue

Objective: PTEN can inhibit the activity of PI3K/AKT signaling and regulate cell proliferation and apoptosis. Increased expression of microRNA-21 is associated with osteosarcoma. Bioinformatics analysis showed a targeted binding site between microRNA-21 and PTEN 3 -UTR. Our study assessed whether microRNA-21 regulates PTEN-PI3K/AKT signaling and affects the proliferation, cloning and apoptosis of osteosarcoma cells. Methods: Dual luciferase reporter gene assay was used to assess the targeted interaction between microRNA-21 and PTEN. Expression of microRNA21 and PTEN was measured in human normal osteoblasts hFOB1.19, osteosarcoma Saos-2 and MG-63. Saos-2 cells were cultured and divided into microRNA-NC group and microRNA-21 inhibitor group followed by measuring the expression of microRNA-21, PTEN and p-AKT, cell apoptosis by flow cytometry, cell proliferation by EdU staining and cloning ability by plate cloning. Results: There was a targeted relationship between microRNA-21 and PTEN. Compared with hFOB1.19 cells, microRNA-21 level in Saos-2 and MG-63 cells was increased and PTEN was decreased. Transfection of microRNA-21 inhibitor significantly reduced microRNA-21 level in Saos-2 cells, increased PTEN, decreased p-AKT, cell proliferation and cloning ability, as well as promoted cell apoptosis. Conclusion: The increased microRNA-21 expression may play a role in reducing PTEN level and promoting osteosarcoma pathogenesis. Inhibiting microRNA-21 can inhibit the activity of PTENPI3K/AKT signaling, reduce the proliferation and cloning ability of osteosarcoma cells, and promote cell apoptosis.

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