1.Insulin as a Potent Stimulator of Akt, ERK and Inhibin-βE Signaling in Osteoblast-Like UMR-106 Cells.
Mahesh RAMALINGAM ; Yong Dae KWON ; Sung Jin KIM
Biomolecules & Therapeutics 2016;24(6):589-594
Insulin is a peptide hormone of the endocrine pancreas and exerts a wide variety of physiological actions in insulin sensitive tissues, such as regulation of glucose homeostasis, cell growth, differentiation, learning and memory. However, the role of insulin in osteoblast cells remains to be fully characterized. In this study, we demonstrated that the insulin (100 nM) has the ability to stimulate the phosphorylation of protein kinase B (Akt/PKB) and extracellular signal-regulated kinase (ERK) and the levels of inhibin-βE in the osteoblast-like UMR-106 cells. This insulin-stimulated activities were abolished by the PI3K and MEK1 inhibitors LY294002 and PD98059, respectively. This is the first report proving that insulin is a potential candidate that enables the actions of inhibin-βE subunit of the TGF-β family. The current investigation provides a foundation for the realization of insulin as a potential stimulator in survival signaling pathways in osteoblast-like UMR-106 cells.
Glucose
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Homeostasis
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Humans
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Insulin*
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Islets of Langerhans
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Learning
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Memory
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Osteoblasts
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Phosphorylation
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Phosphotransferases
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Proto-Oncogene Proteins c-akt
2.Study in the killing effect of Myxoma virus to C6 glioma cell in vitro.
Meng ZANG ; Qiu-Sheng ZHANG ; Shi-Jie LIANG ; Tao JI ; Heng-Zhou LIN ; Wei-Ping LI
Chinese Journal of Experimental and Clinical Virology 2012;26(1):43-45
OBJECTIVETo evaluate the susceptibility of C6 glioma cells to Myxoma virus and the killing effect of Myxoma virus to the C6 glioma cells in vitro.
METHODSC6 glioma cells were infected with myxoma virus, used death virus as the negative control, 5-FU as the positive control, DEMD as blank control. The number of living cells were counted every 24 h, and Western-Blot method, inverted microscope and MTT assay were applicated to observe the cell morphology and survival rate in each group.
RESULTSThe cell number were decreased rapidly in virus effected group and 5-FU group, with significant differences to the negative and blank control groups. And cells in virus effected group appeared cytopathic effect.
CONCLUSIONSC6 glioma cells were susceptible to myxoma virus and myxoma virus had killing effect to C6 glioma cells in vitro.
Cell Line, Tumor ; Glioma ; therapy ; Humans ; Myxoma virus ; Oncolytic Virotherapy ; Proto-Oncogene Proteins c-akt ; physiology
4.Relationship of PTEN/PI3K/AKT Signaling Pathway Protein Expression with Apoptosis and Drug-resistance of Children's ALL Primary Cells Treated with Daunorubicin.
Li YANG ; Jiao-Jiao LI ; Yan-Yang CHENG ; Wen-Jie LU ; Zhuo WANG ; Fang TAO ; Jian-Xin LI ; Zhi-Liang XU ; Hao XIONG
Journal of Experimental Hematology 2019;27(4):1053-1057
OBJECTIVE:
To investigate the relationship of PTEN/PI3K/AKT signaling pathway protein expression with apoptosis and drug-resistance of children's ALL primary cells treated with daunorubicin (DNR).
METHODS:
The bone marrow mononuclear cells in newly diagnosed and untreated B-ALL children were collected and cultured. After the treatment of primary-cultured cells with DNR of final concentration 0.5 mg/L for 24 h, the cell apoptosis rate was detected by using cell apoptosis assay kit; the samples were collected at the on test of culture and after drug treatment, then expression levels of PTEN, PI3K and AKT proteins were detected by Western blot, moreover the interindex correlation was analyzed.
RESULTS:
After DNR treatment, the apoptosis rate in PTEN low expression group was lower than that in PTEN high expression group (P<0.05), showing high positive correlation of the cell apoptosis rate with the expression of PTEN before DNR treatment; the cell apoptosis rate in PI3K and AKT low expression group was higher than that in PI3K and AKT high expression group (P<0.01); however, the expression of PI3K and AKT proteins was down-regulated after treatment with DNR (P<0.01).
CONCLUSION
The difference of PTEN expression is present in primary cells of B-ALL children, however the change of PTEN expression is not significant after DNR treatment, suggesting that the PTEN expression correlates with DNR-resistance. The DNR can induce the apoptosis of childrens B-ALL primary cells by down-regulating the expression of PI3K and AKT signaling pathway proteins.
Apoptosis
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Child
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Daunorubicin
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Humans
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PTEN Phosphohydrolase
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Phosphatidylinositol 3-Kinases
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Proto-Oncogene Proteins c-akt
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Signal Transduction
5.Angelica Polysaccharide Resists Platelets Apoptosis Induced by LY294002.
Hui-Ling WEI ; Chi-Xiang LIU ; Shi-Chao CHEN ; Mo YANG
Journal of Experimental Hematology 2019;27(4):1208-1214
OBJECTIVE:
To investigate the anti-apoptotic effect of Angelica polysaccharide (APS) on cryopreservated platelets and its mechanism.
METHODS:
The platelets were divided into 4 group: control group(4 ℃ stored platelets),APS group (APS-treated platelets stored at 4 ℃), LY294002 group (LY294002-treated platelets stored at 4 ℃) and LY294002+APS group(LY294002+APS treated platelets stored at 4 ℃ ). The expression of platelet membrane glycoprotein CD41 and CD61, as well as the platelet apoptotic rate, Caspase 3 expression and mitochondrial membrane potential (MMP) were detected by flow cytometry; the anti-apoptotic mechanism of APS by PI3K /AKT signaling pathway was analyzed by Western blot assay.
RESULTS:
The apoptosis rate of platelets in LY294002 group obviously increased, the activity of CD41 and CD61 expression gradually decreased along with the enhancement of LY294002 concentrations (r=-0.953); compared with control group, the apoptosis rate of platelets in LY294002 group was enhanced significantly(P<0.05),while the apoptosis rate of platelets in LY294002+APS group significantly was reduced(P<0.05) as compare with LY294002 group, which suggest that APS has an anti-apoptotic effect on the cryopreserved platelets. APS decreased the expression of Caspase-3 and inhibited the reduction of mitochondrial membrane potential induced by LY294002, moreover, APS could increase the activation of PI3K /AKT pathway in Plt.
CONCLUSION
APS has an anti-apoptotic effect on the cryopreserved platelets through activating the PI3K /AKT pathway, decreasing the expression of apoptosis protease Caspase-3 and inhibiting the reduction of MMP.
Angelica
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Apoptosis
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Blood Platelets
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Chromones
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Morpholines
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Phosphatidylinositol 3-Kinases
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Polysaccharides
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Proto-Oncogene Proteins c-akt
6.Costunolide Induces Apoptosis of K562/ADR Cells through PI3K/AKT Pathway.
Journal of Experimental Hematology 2021;29(1):68-71
OBJECTIVE:
To explore the effects of costunolide on the proliferation and apoptosis of human chronic myeloid leukemia drug resisitant cell line K562/ADR and its mechanism.
METHODS:
The proliferation of the cells was assessed by CCK-8 assay, while flow cytometry was used to detect the apoptosis of the cells. The related-proteins were detected by using Western blot.
RESULTS:
The proliferation of K526/ADR cells was significantly inhibited by costunolide in a dose-dependent manner (r=0.9886) after treated by 0.01, 0.1, 0.25, 0.5, 1, 2.5, 5, 10, 25, 50 and 100 μmol/L costunolide for 72 h, and IC
CONCLUSION
Costunolide could inhibit the proliferation and apoptosis of K562/ADR cells through regulation of PI3K/AKT pathway.
Apoptosis
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Cell Proliferation
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Humans
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K562 Cells
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Phosphatidylinositol 3-Kinases
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Proto-Oncogene Proteins c-akt
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Sesquiterpenes
7.Mangiferin attenuates IL
Peng ZHU ; Cong LIU ; Bo LI ; Chen ZHAO ; Tao ZHOU ; Xin XUE ; Bing ZHANG
Journal of Central South University(Medical Sciences) 2021;46(1):25-31
OBJECTIVES:
Chondrocyte apoptosis is an important process in the pathogenesis of osteoarthritis. Mangiferin exerts multiple pharmacological effects such as anti-inflammatory and anti-apoptosis. However, the role of mangiferin in chondrocyte apoptosis is not clear. In this study, we aimed to explore the role of mangiferin in IL-1β-induced chondrocyte apoptosis.
METHODS:
ATDC5 cells were randomly divided into a control group, a IL-1β group, a MFN-L group, a MFN-M group, a MFN-H group and a MFN+LY294002 group. Cells in the control group were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN-L group, the MFN-M group and the MFN-H group were pretreated with 5, 10 and 20 μmol/L mangiferin for 1 h respectively, and then they were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN+LY294002 group were treated with LY294002 (25 μmol/L) for 1 h, then mangiferin (20 μmol/L) and IL-1β (10 ng/mL) for 1 h and 24 h, respectively. Cell viability was detected by CCK-8 assay and cell apoptosis was measured by flow cytometry. Colorimetric assay was conducted to measure the caspase-3 activity. The protein levels of Bcl-2, Bax, and phosphoinositide 3-kinase (PI3K)/Akt signaling pathway related proteins were detected by Western blotting.
RESULTS:
Compared to the control group, cell viability was significantly decreased; cell apoptosis, caspase-3 activity and Bax protein expression were significantly increased; the protein levels of Bcl-2, p-PI3K, and p-Akt were significantly decreased in the IL-1β group (all
CONCLUSIONS
Mangiferin could attenuate IL-1β-induced apoptosis of the mice chondrocytes, which is mediated by the activation of PI3K/Akt signaling pathway.
Animals
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Apoptosis
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Chondrocytes
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Interleukin-1beta
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Mice
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Phosphatidylinositol 3-Kinases
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Proto-Oncogene Proteins c-akt
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Xanthones
8.Role of autophagy in lipopolysaccharide-induced apoptosis of odontoblasts.
Huacui XIONG ; Ke CHEN ; Meimei LI
Journal of Southern Medical University 2020;40(12):1816-1820
OBJECTIVE:
To investigate the role of autophagy in lipopolysaccharide (LPS)-induced apoptosis of murine odontoblasts.
METHODS:
Murine odontoblasts (mDPC-23 cells) were treated with 5 μg/mL LPS for 6, 12 and 24 h, and the changes in cell viability was examined using CCK8 kit and cell apoptosis was detected by TUNEL staining. The changes in the protein levels of LC3, Beclin1, Atg5, AKT, p-AKT, mTOR and p-mTOR were detected using Western blotting. The effect of 3-MA treatment for 24 h on LPS-induced apoptosis of mDPC-23 cells was evaluated by detecting the expressions of apoptosis-related proteins caspase-3 and Bax using Western blotting.
RESULTS:
Stimulation with LPS for 6 and 12 h did not cause significant changes in the proliferation or apoptosis of mDPC-23 cells, but LPS treatment for 24 h significantly suppressed cell proliferation (
CONCLUSIONS
LPS stimulation induces autophagy to promote apoptosis of mDPC-23 cells, and suppression of autophagy attenuates LPS-induced apoptosis. Autophagy may play an important role in the injury of inflamed pulp tissues.
Animals
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Apoptosis
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Autophagy
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Lipopolysaccharides/pharmacology*
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Mice
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Odontoblasts/metabolism*
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Proto-Oncogene Proteins c-akt/metabolism*
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Signal Transduction
9.Effect of Xuanfu Daizhe decoction on stemness of esophageal cancer cells.
Yuan MA ; Jing XUN ; Bo-Tao WANG ; Di-Hua LI ; Qi ZHANG ; Zhen-Yu WANG ; Yu WU
Chinese Journal of Applied Physiology 2022;38(1):62-67
Objective: To investigate the effect of Xuanfu Daizhe decoction on the stemness of esophageal cancer cells. Methods: The BALB/c nude mice were randomly divided into the control group and experimental group, 5 mice in each group, which were continuously administered with normal saline and Xuanfu Daizhe decoction (9.89 g/kg) by gastrogavage, respectively. Human esophageal carcinoma cells ECA-109 (5×106) were subcutaneously injected into the mice on the 8th day. Tumor volume was measured twice a week. The mice were sacrificed 4 weeks after injection, and the tumor tissue and mouse serum were collected. The expressions of the major stemness-regulating transcription factors, i.e., NANOG, OCT4 and SOX2, were detected by RT-qPCR, Western Blot and immunohistochemistry. ECA-109 cells were treated with 10% fetal bovine serum and serum from the above two groups of mice for 48 hours respectively, and three replicate wells were set in each group, and the expressions of NANOG, OCT4, SOX2 and the levels of AKT and p-AKT were detected by RT-qPCR and Western Blot, respectively. ALDH activity in tumor cells was detected by flow cytometry; the number of spheroids of tumor cells was detected by the spheroidization experiment. Results: Compared with the control group, the growth and size of esophageal cancer tumors were significantly inhibited by Xuanfu Daizhe Decoction; the expressions of NANOG, OCT4, SOX2, the ALDH activity, the number of spheroids, and the levels of AKT and phosphorylated AKT (p-AKT) in esophageal cancer cells were significantly reduced by Xuanfu Daizhe Decoction both in vivo and in vitro. Conclusion: Xuanfu Daizhe Decoction inhibits the stemness of esophageal cancer cells, it may be a potentially effective drug for the treatment of esophageal cancer and provides a theoretical basis for the exploration of new effective drugs for the treatment of esophageal cancer.
Animals
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Esophageal Neoplasms/pathology*
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Mice
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Mice, Nude
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Proto-Oncogene Proteins c-akt
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Transcription Factors
10.Piroctone olamine disrupts mitochondrial dynamics in glioma cells through the PI3K/AKT pathway.
Wenqin XU ; Jingjing YE ; Fei WANG ; Tianbing CHEN
Journal of Southern Medical University 2023;43(5):764-771
OBJECTIVE:
To investigate the growth-inhibitory and pro-apoptotic effects of piroctone olamine (PO) on glioma cells and explore the underlying mechanism.
METHODS:
Human glioma cell lines U251 and U373 were treated with PO and the changes in cell proliferation were detected using CCK-8 assay and EdU assay. Clone formation assay and flow cytometry were used to examine the changes in clone formation ability and apoptosis of the treated cells. Mitochondrial membrane potential of the cells and morphological changes of the mitochondria were detected using JC-1 staining and a fluorescence probe, respectively. The expressions of mitochondrial fission protein DRP1 and the fusion protein OPA1 were determined with Western blotting. Transcriptome sequencing and differential gene enrichment analysis was performed, and the expression levels of PI3K, AKT and p-AKT in the treated cells were verified using Western blotting.
RESULTS:
CCK-8 assay showed that PO significantly inhibited the proliferation of U251 and U373 cells in a time- and dose-dependent manner (P < 0.001). EdU test showed that the proliferative activity of PO-treated cells was significantly decreased, and the number of cell colonies also decreased significantly (P < 0.01). PO treatment significantly increased apoptotic rates (P < 0.01), decreased mitochondrial membrane potential and caused obvious changes in mitochondrial morphology of the cells. Pathway enrichment analysis showed that the down-regulated genes were significantly enriched in the PI3K/AKT pathway, which was verified by Western blotting showing significantly down-regulated expression levels of PI3K, AKT and p-AKT in PO-treated cells (P < 0.05).
CONCLUSION
PO interferes with mitochondrial fusion and fission function through the PI3K/AKT pathway, thereby inhibiting the proliferation and increasing apoptosis of glioma cells.
Humans
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Glioma
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Mitochondrial Dynamics
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Phosphatidylinositol 3-Kinases
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Proto-Oncogene Proteins c-akt