Amyloid peptide, the main component of senile plaques, is a major biological characteristic of Alzheimer’s disease (AD). The aim of the present study conducted on human neuronal SK-N-BE cells was to evaluate whether ...Amyloid peptide, the main component of senile plaques, is a major biological characteristic of Alzheimer’s disease (AD). The aim of the present study conducted on human neuronal SK-N-BE cells was to evaluate whether oligomerized Aβ1-40-induced cell damages was associated with lipid modifications. Under treatment with Aβ1-40 (10 - 100 μM;24 - 48 h), cell viability was recorded with the MTT test and by measuring LDH activity. Mitochondrial transmembrane potential and ATP production were assessed using flow cytometry and a luciferase-based ATP bioluminescence assay, respectively. Annexin V-CF647 staining assay for cell apoptosis detection was performed using flow cytometry. Potentially intracellular cytotoxic lipids (oxysterols: 7α-hydroxycholesterol (7α-OHC), 7β-hydroxycholesterol (7β-OHC), and 7-ketocholesterol (7KC), 24(S)-hydroxycholesterol;arachidonic acid (C20:4 n-6);VLCFAs (C22:0, C24:0, C24:6 and C26:0)) were measured using gas chromatography coupled with mass spectrometry. The cellular level of docosahexaenoic acid (C22:6 n-3), often altered in AD, was also quantified. In the presence of Aβ1-40, the percentage of MTT-positive cells decreased and was associated with an increase in LDH activity. In addition, treatment with oligomerized Aβ1-40 induced a decrease of mitochondrial transmembrane potential as well as an apoptotic cell death. Sterol analysis revealed a higher cholesterol level and a significant increase of cytotoxic oxysterols per cell (7KC + 7β-OHC), and of the [(7β-OHC + 7KC)/cholesterol] ratio, considered as a lipid peroxidation index, in Aβ1-40-treated cells. An enhancement of C20:4 n-6, C22:6 n-3 and saturated VLCFAs was also observed. Therefore, Aβ1-40-induced side effects are associated with intracellular accumulation of lipids, especially cholesterol, oxysterols (7β-OHC, 7KC), C20:4 n-6, and saturated VLCFAs, which could in turn contribute to neurotoxicity.展开更多
Objective This study aims to investigate the effects of the long noncoding RNA(lncRNA)DRAIC on the proliferation,apoptosis,and radiosensitivity of hepatocellular carcinoma(HCC)cells and the molecular mechanisms involv...Objective This study aims to investigate the effects of the long noncoding RNA(lncRNA)DRAIC on the proliferation,apoptosis,and radiosensitivity of hepatocellular carcinoma(HCC)cells and the molecular mechanisms involved.Methods Cancer tissues and their corresponding adjacent tissues from 30 patients with HCC were collected,and the expression levels of DRAIC and miR-223-3p were detected via RT-q PCR.DRAIC interference and miR-223-3p overexpression vectors were transfected into HepG2 cells.In addition,DRAIC and miR-223-3p interference vectors were co-transfected into HepG2 cells.The constructed cells were irradiated at 4 Gy.Cell colony formation assay,MTT assay,and flow cytometry were performed to detect the radiosensitivity,proliferation inhibition rate,and apoptosis rate of HepG2 cells,respectively.Dual luciferase reporter gene assay was performed to detect the targeted regulation of DRAIC on miR-223-3p expression.Results The expression level of DRAIC in HCC tissues was higher than that in paracancer tissues,whereas the expression level of miR-223-3p was lower in HCC tissues than that in paracancer tissues(P<0.05).Inhibition of DRAIC expression or overexpression of miR-223-3p increased the proliferation inhibition and apoptosis rates of HepG2 cells(P<0.05).After irradiation,cell survival fraction decreased and cell proliferation inhibition and apoptosis rates increased(P<0.05).DRAIC targeted the regulation of miR-223-3p expression,and interference of miR-223-3p expression reversed the effects of inhibiting DRAIC expression on the proliferation,apoptosis,and radiosensitivity of HepG2 cells.Conclusion Inhibition of DRAIC expression can inhibit the proliferation of HepG2 cells,promote cell apoptosis,and enhance the radiosensitivity of cells via upregulation of miR-223-3p.展开更多
文摘Amyloid peptide, the main component of senile plaques, is a major biological characteristic of Alzheimer’s disease (AD). The aim of the present study conducted on human neuronal SK-N-BE cells was to evaluate whether oligomerized Aβ1-40-induced cell damages was associated with lipid modifications. Under treatment with Aβ1-40 (10 - 100 μM;24 - 48 h), cell viability was recorded with the MTT test and by measuring LDH activity. Mitochondrial transmembrane potential and ATP production were assessed using flow cytometry and a luciferase-based ATP bioluminescence assay, respectively. Annexin V-CF647 staining assay for cell apoptosis detection was performed using flow cytometry. Potentially intracellular cytotoxic lipids (oxysterols: 7α-hydroxycholesterol (7α-OHC), 7β-hydroxycholesterol (7β-OHC), and 7-ketocholesterol (7KC), 24(S)-hydroxycholesterol;arachidonic acid (C20:4 n-6);VLCFAs (C22:0, C24:0, C24:6 and C26:0)) were measured using gas chromatography coupled with mass spectrometry. The cellular level of docosahexaenoic acid (C22:6 n-3), often altered in AD, was also quantified. In the presence of Aβ1-40, the percentage of MTT-positive cells decreased and was associated with an increase in LDH activity. In addition, treatment with oligomerized Aβ1-40 induced a decrease of mitochondrial transmembrane potential as well as an apoptotic cell death. Sterol analysis revealed a higher cholesterol level and a significant increase of cytotoxic oxysterols per cell (7KC + 7β-OHC), and of the [(7β-OHC + 7KC)/cholesterol] ratio, considered as a lipid peroxidation index, in Aβ1-40-treated cells. An enhancement of C20:4 n-6, C22:6 n-3 and saturated VLCFAs was also observed. Therefore, Aβ1-40-induced side effects are associated with intracellular accumulation of lipids, especially cholesterol, oxysterols (7β-OHC, 7KC), C20:4 n-6, and saturated VLCFAs, which could in turn contribute to neurotoxicity.
文摘Objective This study aims to investigate the effects of the long noncoding RNA(lncRNA)DRAIC on the proliferation,apoptosis,and radiosensitivity of hepatocellular carcinoma(HCC)cells and the molecular mechanisms involved.Methods Cancer tissues and their corresponding adjacent tissues from 30 patients with HCC were collected,and the expression levels of DRAIC and miR-223-3p were detected via RT-q PCR.DRAIC interference and miR-223-3p overexpression vectors were transfected into HepG2 cells.In addition,DRAIC and miR-223-3p interference vectors were co-transfected into HepG2 cells.The constructed cells were irradiated at 4 Gy.Cell colony formation assay,MTT assay,and flow cytometry were performed to detect the radiosensitivity,proliferation inhibition rate,and apoptosis rate of HepG2 cells,respectively.Dual luciferase reporter gene assay was performed to detect the targeted regulation of DRAIC on miR-223-3p expression.Results The expression level of DRAIC in HCC tissues was higher than that in paracancer tissues,whereas the expression level of miR-223-3p was lower in HCC tissues than that in paracancer tissues(P<0.05).Inhibition of DRAIC expression or overexpression of miR-223-3p increased the proliferation inhibition and apoptosis rates of HepG2 cells(P<0.05).After irradiation,cell survival fraction decreased and cell proliferation inhibition and apoptosis rates increased(P<0.05).DRAIC targeted the regulation of miR-223-3p expression,and interference of miR-223-3p expression reversed the effects of inhibiting DRAIC expression on the proliferation,apoptosis,and radiosensitivity of HepG2 cells.Conclusion Inhibition of DRAIC expression can inhibit the proliferation of HepG2 cells,promote cell apoptosis,and enhance the radiosensitivity of cells via upregulation of miR-223-3p.
