Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 is correlated with NSCLC metastasis and could promote NSCLC cell migration in vitro . (A) Representative IHC images of PFN1 expression on the NSCLC tissues. (B) The staining index of PFN1 on the tissue chip. ** p < 0.01. (C) Representative IHC images of PFN1 expression on the tissue chip. (D) The expression of PFN1 in TCGA LUAD data. ** p < 0.01. (E) The Kaplan–Meier survival analysis of PFN1 in NSCLC patients. (Data source: TCGA LUAD dataset) (F,G) Wound healing assays conducted to evaluate the migration ability of PFN1 -overexpressing (F) and PFN1 knockdown (KD) (G) H1299 cells. ** p < 0.01; scale bar, 500 μm. (H,I) Transwell migration assays conducted to evaluate the migration of PFN1 -overexpressing (H) and PFN1 KD (I) H1299 cells. ** p < 0.01; scale bar, 500 μm. EV, empty vector; OE, PFN1 overexpression; NC, negative control; si-1/ 2, PFN1 siRNA1 1/2.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration, In Vitro, Expressing, Staining, Knockdown, Plasmid Preparation, Over Expression, Negative Control

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 could promote MVs secretion in NSCLC. (A) Heatmap of differentially expressed proteins between EV and PFN1 OE cells. (B) GO enrichment analysis of differentially expressed proteins. (C) COG/KOG analysis of differentially expressed proteins. (D) MVs extracted from EV-expressing and PFN1 -overexpressing cells, using continuous differential centrifugation, identified using transmission electron microscopy. Scale bar, 100 nm. (E,F) Flow cytometry (E) and western blotting (F) were used to quantify MVs in PFN1 -overexpressing and EV-expressing cells. ARF6 and actin were used as MV markers. (G) Expression of PFN1 and annexin A1 in lung tumor tissues detected using immunofluorescence. (H) The staining index of p-MLC on the tissue chip. ** p < 0.01. (I) Representative IHC images of p-MLC expression. (J) Spearman rank correlation analysis was used to assess the relationship between PFN1 and p-MLC expression on the tissue chip; p and r values are shown in the plot.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Expressing, Centrifugation, Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: MVs derived from PFN1 OE cells promote migration in NSCLC cells. (A) MVs collected from sera of patients with NSCLC quantified using flow cytometry. ** p < 0.01. (B) Protein expression of ARF6 and β-actin in MVs collected from sera of patients with NSCLC detected using western blotting. (C) Effect of PFN1 -overexpressing cell supernatants on cell migration evaluated through wound healing assays. ** p < 0.01; scale bar, 500 μm. (D) PKH67-labeled MVs taken up by H1299 cells. DAPI was used to stain the nuclei of H1299 cells. Scale bar, 500 μm. (E,F) Wound healing (E) and Transwell migration (F) assays conducted to evaluate the migration of H1299 cells after treatment with MVs derived from EV-expressing and PFN1 -overexpressing cells; ** p < 0.01; scale bar, 500 μm.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Derivative Assay, Migration, Flow Cytometry, Expressing, Western Blot, Labeling, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 promotes in vivo NSCLC metastasis by elevating MV secretion. (A) Schematic illustration of the mouse model of metastatic tumor established to determine the role of PFN1 in tumor metastasis. (B) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing and EV-expressing cell lines. (C,D) Representative images of lung (C) and liver (D) metastases of the mouse model. The number of metastases is displayed in the right-hand side graph. * p < 0.05, ** p < 0.01. (E) Representative images of HE-stained lung tissues of the mouse model. (F) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (G) Representative images of HE-stained liver tissues of the mouse model. (H) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (I) Body weight changes in mice after intracardiac injection of H1299 cells and MVs. (J) Representative images of lung metastases of the mouse model. The number of metastases is shown in the bottom graph. * p < 0.05. (K) Representative images of HE-stained lung tissues of the mouse model. (L) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vivo, Injection, Expressing, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Mechanisms underlying the promotion of MLC phosphorylation by PFN1. (A,B) Protein expression after PFN1 overexpression (A) and knockdown (B) measured using western blotting. (C) Protein expression in PFN1 mutants measured using western blotting. (D) PFN1 interactions with ROCK1/2 confirmed using co-IP. (E) Protein expression after treatment with Y27632 (10 µM) measured using western blotting. (F) Effect of PFN1 on ROCK1 activity. ** p < 0.01. (G) Effect of PFN1 on ROCK2 activity. (H) Flow cytometry measuring changes in the amount of MVs after treatment with Y27632; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Phospho-proteomics, Expressing, Over Expression, Knockdown, Western Blot, Co-Immunoprecipitation Assay, Activity Assay, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: ROCK1 inhibitor Y27632 partially reversed the promotion of lung cancer metastasis by PFN1 in vitro and in vivo . (A,B) Wound healing assays conducted to evaluate the effect of Y27632 (A) and Y27632 combined with MVs (B) on cell migration. ** p < 0.01; scale bar, 500 μm. (C) Transwell migration assays conducted to evaluate the effect of Y27632 and Y27632 combined with MVs on cell migration. ** p < 0.01; scale bar, 500 μm. (D) Schematic diagram of the mouse model of metastatic tumor established to determine the effect of Y27632 on PFN1-induced lung cancer metastasis. (E) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing H1299 cells and intraperitoneal injection of Y27632 (10 mg/kg). (F) Representative images of lung and liver metastatic tissue in mice. The number of metastatic nodules is shown in the right-hand side graph. * p < 0.05. (G,H) Representative images of HE-stained lung (G) and liver (H) metastases. (I) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (J) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph; ** p < 0.01.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vitro, In Vivo, Migration, Injection, Staining, Expressing

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Schematic diagram of the role of PFN1 in NSCLC metastasis. In the initiation stage of NSCLC, cells with upregulated PFN1 secret more MVs through PFN1 interactions with the ROCK/p-MLC pathway. These MVs contain numerous oncogenenic moleculars, which could enhance migration abilities of PFN1 normal expressed NSCLC cells, and untimately promote progression and metastasis of NSCLC.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-150 suppresses the growth and malignant behavior of papillary thyroid carcinoma cells via downregulation of MUC4

doi: 10.3892/etm.2018.6197

Figure Lengend Snippet: Upregulation of miR-150 suppresses human papillary thyroid carcinoma cell proliferation and metastasis. (A) Relative miR-150 expression in TPC-1 cells following transfection. (B) Proliferation of TPC-1 cells following transfection. (C) Representative micrographs of a wound-healing assay in TPC-1 cells following transfection (Scale bars=50 µm). (D) Representative micrographs of Transwell invasion experiments in TPC-1 cells following transfection (Scale bars=50 µm). (E) Statistical analysis of the wound-healing assay results. (F) Statistical analysis of the Transwell invasion assay results. *P<0.05 vs. mimic-Con, **P<0.01 vs. mimic-Con. miR-150, microRNA-150; mimic-Con, negative control transfection; OD, optical density.

Article Snippet: miR-150 mimics (5′-UCUCCCAACCCUUGUACCAGUG-3′) and negative control miR sequences (5′-CCGAAACCUCGGUUGAUUGCGG-3′) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Transfection, Wound Healing Assay, Transwell Invasion Assay, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-150 suppresses the growth and malignant behavior of papillary thyroid carcinoma cells via downregulation of MUC4

doi: 10.3892/etm.2018.6197

Figure Lengend Snippet: MUC4 is a direct target of miR-150 in papillary thyroid carcinoma cells. (A) miR-150 and its target site in MUC4 3′-UTR. The mutant bases are indicated by boxes. (B) Luciferase reporter assay in TPC-1 cells transfected with reporter vectors containing WT or MUT MUC4 3′-UTR. (C) MUC4 mRNA expression was evaluated by reverse transcription-quantitative polymerase chain reaction in TPC-1 cells transfected with miR-150 mimics and mimic-Con. (D) MUC4 protein expression was evaluated by western blot analysis in TPC-1 cells transfected with miR-150 mimics and mimic-Con. (E) Densitometric analysis of MUC4 protein level. *P<0.05 vs. control; **P<0.01 vs. mimic-Con. miR-150, microRNA-150; mimic-Con, negative control transfection; MUC4, mucin 4; 3′-UTR, 3′-untranslated region; NC, negative control; WT, wild type; MUT, mutated.

Article Snippet: miR-150 mimics (5′-UCUCCCAACCCUUGUACCAGUG-3′) and negative control miR sequences (5′-CCGAAACCUCGGUUGAUUGCGG-3′) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Mutagenesis, Luciferase, Reporter Assay, Transfection, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-150 suppresses the growth and malignant behavior of papillary thyroid carcinoma cells via downregulation of MUC4

doi: 10.3892/etm.2018.6197

Figure Lengend Snippet: Effect of miR-150 on HER2 and downstream FAK/ERK signaling. (A) Protein levels were evaluated by western blot analysis. (B) Densitometric analysis of each protein level. **P<0.01 vs. mimic-Con. miR-150, microRNA-150; mimic-Con, negative control transfection; HER2, human epidermal growth factor receptor 2; FAK, focal adhesion kinase; ERK, extracellular signal-regulated kinase.

Article Snippet: miR-150 mimics (5′-UCUCCCAACCCUUGUACCAGUG-3′) and negative control miR sequences (5′-CCGAAACCUCGGUUGAUUGCGG-3′) were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Negative Control, Transfection

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