Journal: Journal of Translational Medicine

Article Title: Artemisinin conferred cytoprotection to human retinal pigment epithelial cells exposed to amiodarone-induced oxidative insult by activating the CaMKK2/AMPK/Nrf2 pathway

doi: 10.1186/s12967-024-05593-x

Figure Lengend Snippet: The Nrf2 pathway was involved in the cytoprotective effect of artemisinin on amiodarone-induced cell death in D407 cell cultures. ( a-b ) The cells were treated with artemisinin at various concentrations for 1 h. The Nrf2 and GAPDH protein levels were detected by western blotting, and protein bands were quantified by Image J ( n = 3). ( c ) The cell cultures were incubated with or without the Nrf2 inhibitor ML385 at concentrations ranging from 2.5 to 15 µM for 24 h, and cell viability was measured using MTT ( n = 4). ( d-e ) Cells attached to 12-well plates were treated with 5 or 10 µM ML385 for 24 h. The Nrf2 and GAPDH protein levels were detected by western blotting, and protein bands were quantified by Image J ( n = 4). ( f-g ) The cell cultures were pretreated with 5 µM ML385 for 6, 12, or 24 h, followed by 20 µM artemisinin for 1 h and 5 µM amiodarone for another 24 h. Cell viability was determined by the MTT assay ( n = 4), and cell death was detected through the LDH cytotoxicity assay ( n = 4). ( h-j ) Cells attached to 24-well plates were transfected with control shRNA (shCTL) or Nrf2 shRNA (shNrf2) plasmids (0.5 µg/well) for 72 h. The Nrf2, HO-1, and GAPDH protein levels were detected by western blotting, and protein bands were quantified by Image J ( n = 3). ( k ) Cells transfected with shCTL or shNrf2 plasmids were attached to 96-well plates, and pretreated with artemisinin for 1 h, then treated with amiodarone for another 24 h and processed for LDH assay ( n = 4)

Article Snippet: The human AMPKɑ2 shRNA plasmids (target sequence: TGTGAAAGAAGTGTGTGAA), control shRNA plasmids (CON207, target sequence: TTCTCCGAACGTGTCACGT), the AMPKɑ2-overexpressing plasmids, and control overexpressing plasmids (CON238) were purchased from GenePharma Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Incubation, MTT Assay, LDH Cytotoxicity Assay, Transfection, Control, shRNA, Lactate Dehydrogenase Assay

Journal: Journal of Translational Medicine

Article Title: Artemisinin conferred cytoprotection to human retinal pigment epithelial cells exposed to amiodarone-induced oxidative insult by activating the CaMKK2/AMPK/Nrf2 pathway

doi: 10.1186/s12967-024-05593-x

Figure Lengend Snippet: AMPK-mediated activation of the Nrf2/HO-1 pathway was linked to the cytoprotective effect of artemisinin on amiodarone-induced apoptosis in D407 cell cultures. ( a-d ) The cell cultures were pretreated with 1 µM STO-609 for 30 min or 1 µM Compound C for 30 min, followed by 20 µM artemisinin for 1 h and 5 µM amiodarone for another 24 h. The HO-1, cleaved caspase-3, and GAPDH protein levels were detected by western blotting, and the protein bands were quantified by Image J ( n = 3). ( e-h ) Cells attached to 12-well plates were transfected with shCTL or AMPKɑ2 shRNA (shAMPKɑ2) plasmids (2 µg/well) for 48 h. The AMPKɑ2, HO-1, cleaved caspase-3, and GAPDH protein levels were detected by western blotting, and protein bands were quantified by Image J ( n = 3). ( i-j ) Cells attached to 24-well plates were transfected with the AMPKɑ2-overexpressing (OE-AMPKɑ2) or control-overexpressing (OE-CTL) plasmids (0.5 µg/well) for 24 h. The AMPKɑ2 and GAPDH protein levels were detected by western blotting, and protein bands were quantified by Image J ( n = 3). ( k ) The cell cultures transfected with OE-CTL or OE-AMPKɑ2 plasmids for 24 h were attached to 96-well plates and treated with 5 µM amiodarone for 24 h, and processed for the LDH assay ( n = 4)

Article Snippet: The human AMPKɑ2 shRNA plasmids (target sequence: TGTGAAAGAAGTGTGTGAA), control shRNA plasmids (CON207, target sequence: TTCTCCGAACGTGTCACGT), the AMPKɑ2-overexpressing plasmids, and control overexpressing plasmids (CON238) were purchased from GenePharma Co., Ltd. (Shanghai, China).

Techniques: Activation Assay, Western Blot, Transfection, shRNA, Control, Lactate Dehydrogenase Assay

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Flow chart of the derivation and function analysis of stress associated endoplasmic reticulum protein 1 (SERP1). (A) Flow chart. (B) Heat map of differentially expressed mRNAs in pancreatic ductal adenocarcinoma (PDAC) from GEO profiles database. The 10 mRNAs with the most significant difference are shown.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques:

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Expression level of the stress associated endoplasmic reticulum protein 1 (SERP1) in pancreatic ductal adenocarcinoma (PDAC) tissues is upregulated. (A) SERP1 was upregulated in six pairs of PDAC tissues compared with adjacent normal tissues. (B) The expression of SERP1 in eight pairs of PDAC tissues was upregulated compared with adjacent normal tissues from GEO database. (C and D) The Human Protein Atlas database results revealed that the expression level of SERP1 were upregulated in PDAC tissues compared with normal tissues. (E) Amplification of SERP1 was found in various tumors, including PDAC.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Expressing, Amplification

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Correlation between expression level of the stress associated endoplasmic reticulum protein 1 (SERP1) and progression of pancreatic ductal adenocarcinoma (PDAC) patients. (A and B) The expression level of SERP1 in T3/4 stage PDAC specimen was significantly higher than in T1/2 stage, while there was no significant difference between SERP1 and N stage. (C) Correlation analysis was performed between expression level of SERP1 and clinical stages. SERP1 was correspondingly increased with the increase of clinical AJCC stage of PDAC. (D) Constituent ratio with high level of SERP1 was drastically different between normal samples and I/II or III/IV stage of PDAC patients.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: The correlation between expression level of the stress associated endoplasmic reticulum protein 1 (SERP1) and prognosis of pancreatic ductal adenocarcinoma (PDAC) patients. (A) K-M curve for OS of patients with high (n=89) and low (n=87) SERP1 expression level. Results revealed that high SERP1 expression level was correlated with shorter OS for PDAC patients. (B) SERP1 high expression group (n=61) had shorter DFS than SERP1 low expression group (n=76). (C) The expression of SERP1 in OS-good and poor patients. (D) The expression of SERP1 in DFS-good and poor patients. (E and F) Multivariate Cox regression analyses showed SERP1 expression was an independent factor of patients' overall survival and disease-free survival.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Stress associated endoplasmic reticulum protein 1 (SERP1) is mainly involved in regulating cell apoptosis. (A) Apoptosis gene sets were enriched in tumor group. The curve is the enrichment score of these genes by GSEA. (B) PANC-1 cells were transfected with three SERP1 siRNA. Forty-eight hours after transfection, the expression level of SERP1 was detected by western blotting, and results revealed that the expression level of SERP1 was obviously downregulated in PANC-1 cells transfected with SERP1-siRNA compared to negative control cells. (C and D) Compared to control group, PANC-1 cells transfected with SERP1-siRNA had a higher rate of apoptosis.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Transfection, Expressing, Western Blot, Negative Control, Control

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Protein-protein interaction (PPI) and GO analyses for interaction genes with the stress associated endoplasmic reticulum protein 1 (SERP1). (A and B) PPI analysis of interaction genes with SERP1. SRP receptor β subunit (SRPRB) was the main node protein for the interaction network. (C) GO analysis of the interaction gene with SERP1, and these genes were mainly involved in regulating cell death and apoptosis.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques:

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Co-expression genes with SERP1 from cBioPortal database.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques:

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Downregulated stress associated endoplasmic reticulum protein 1 (SERP1) promotes apoptosis via increasing SRP receptor β subunit (SRPRB) expression. (A and B) The Human Protein Atlas database results revealed that the expression level of SRPRB were upregulated in stromal fibroblast compared with tumor tissues. (C) Western blot analysis confirmed that downregulated SERP1 can increase the expression of SRPRB in PANC-1 cells. (D) PANC-1 cells were transfected with SRPRB overexpression plasmid, and western blot analysis found that the expression level of SRPRB was significantly increased in PANC-1 cells. (E and F) Compared to control group, SRPRB overexpression significantly promoted apoptosis of PANC-1 cells.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Expressing, Western Blot, Transfection, Over Expression, Plasmid Preparation, Control

Journal: International Journal of Oncology

Article Title: SERP1 is a novel marker of poor prognosis in pancreatic ductal adenocarcinoma patients via anti-apoptosis and regulating SRPRB/NF-κB axis

doi: 10.3892/ijo.2017.4111

Figure Lengend Snippet: Downregulated stress associated endoplasmic reticulum protein 1 (SERP1) promotes cell apoptosis via regulating SRP receptor β subunit (SRPRB) associated NF-κB activation. (A) NF-κB signaling pathway gene sets were enriched in the tumor group. (B) Correlation analysis revealed that the expression level of SERP1 positively correlated with NF-κB expression in 178 pancreatic ductal adenocarcinoma (PDAC) patients from TCGA database. (C) The expression level of SRPRB was negatively correlated with NF-κB expression in 178 PDAC patients from TCGA database. (D) Western blotting confirmed that downregulated SERP1 could suppress the expression of NF-κB and the phosphorylation NF-κB in PANC-1 cells.

Article Snippet: The small interfering RNA targeting SERP1 and its negative control (NC) sequences were designed and synthesized by GenePharma Biotech, Co., Ltd. (Shanghai, China).

Techniques: Activation Assay, Expressing, Western Blot, Phospho-proteomics