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Santa Cruz Biotechnology nm 174936
Nm 174936, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nm+174936/PCSK9+siRNA/pmc04440094-87-17-9
Average 93 stars, based on 13 article reviews

nm 174936 - by Bioz Stars, 2026-09

93/100 stars

Images

Transfection:

Article Title: PCSK9 regulates apoptosis in human neuroglioma u251 cells via mitochondrial signaling pathways
Article Snippet: Cells were routinely grown in Dulbecco’s Modified Eagle Media (DMEM, Hyclone, Beijing, China) contained with 10% fetal bovine serum (FBS, Hyclone), 100 U/mL penicillin (Sigma, MO, USA) and 100 μg/mL streptomycin (Sigma) at 37°C in a humidified atmosphere of 5% CO 2 , and were fed every 2-3 d and were split 1 to 2 at 80% confluence. .. U251 cells were transfected with 100 nM PCSK9 siRNA (Santa Cruz, sc-45482; Genbank ID for PCSK9: {"type":"entrez-nucleotide","attrs":{"text":"NM_174936","term_id":"299523249","term_text":"NM_174936"}} NM_174936 ) or control siRNA (scrambled siRNA, a universal negative control) or pcDNA3-PCSK9 with Lipofectamine2000 (Invitrogen), according to the manufacturer’s instructions. ..

Control:

Negative Control:

Image Search Results

Schematic graph presenting the crucial role of proprotein convertase subtilisin/kexin type 9 (PCSK9) on LDL receptor turnover in hepatocytes (created with BioRender.com , accessed on 15 November 2023).

Journal: International Journal of Molecular Sciences

Article Title: The Application of Peptide Nucleic Acids (PNA) in the Inhibition of Proprotein Convertase Subtilisin/Kexin 9 ( PCSK9 ) Gene Expression in a Cell-Free Transcription/Translation System

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Schematic graph presenting the crucial role of proprotein convertase subtilisin/kexin type 9 (PCSK9) on LDL receptor turnover in hepatocytes (created with BioRender.com , accessed on 15 November 2023).

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques:

Integrated optical density (IOD) of DNA bands after conventional reverse transcription PCR (RT–PCR) ( B ) and agarose gel electrophoresis ( A ). An equal amount of RNA taken from the T7 RNA polymerase reaction served as a template. The control sample contained a PNA oligomer that was not complementary to the PCSK9 coding sequence. This sample was arbitrarily assigned a value of 100%. In Panel ( B ), dashed line bars represent real-time PCR, while white dotted black bars represent conventional PCR after 15 cycles, in comparison to the control PNA. *— p < 0.05 vs. ctrlPNA for RT-PCR, #— p < 0.05 vs. ctrlPNA for conventional PCR.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Integrated optical density (IOD) of DNA bands after conventional reverse transcription PCR (RT–PCR) ( B ) and agarose gel electrophoresis ( A ). An equal amount of RNA taken from the T7 RNA polymerase reaction served as a template. The control sample contained a PNA oligomer that was not complementary to the PCSK9 coding sequence. This sample was arbitrarily assigned a value of 100%. In Panel ( B ), dashed line bars represent real-time PCR, while white dotted black bars represent conventional PCR after 15 cycles, in comparison to the control PNA. *— p < 0.05 vs. ctrlPNA for RT-PCR, #— p < 0.05 vs. ctrlPNA for conventional PCR.

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Sequencing, Real-time Polymerase Chain Reaction, Comparison

Western blot image of an in vitro transcription/translation reaction product resolved by electrophoresis with appropriate integrated optical density (I.O.D) values ( A ) and the relative amount of protein with respect to control sample ( B ). 1–4: Samples that contained InitPNA, Ex1PNA, Ex2PNA, and control (non-PCSK9 specific) PNA, respectively. M: Color Prestained Protein Standard, Broad Range (New England Biolabs, no. Cat. P7712). The PCSK9-specific band is depicted with an arrow. A total amount of 10 µg (25 µL sample volume) of purified products from the transcription/translation reaction was loaded onto the gel. The initial protein concentration in the samples was assessed spectrophotometrically. Measurements were made using ImageJ 1.53t software [ , ] *: p < 0.05 vs. control.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Western blot image of an in vitro transcription/translation reaction product resolved by electrophoresis with appropriate integrated optical density (I.O.D) values ( A ) and the relative amount of protein with respect to control sample ( B ). 1–4: Samples that contained InitPNA, Ex1PNA, Ex2PNA, and control (non-PCSK9 specific) PNA, respectively. M: Color Prestained Protein Standard, Broad Range (New England Biolabs, no. Cat. P7712). The PCSK9-specific band is depicted with an arrow. A total amount of 10 µg (25 µL sample volume) of purified products from the transcription/translation reaction was loaded onto the gel. The initial protein concentration in the samples was assessed spectrophotometrically. Measurements were made using ImageJ 1.53t software [ , ] *: p < 0.05 vs. control.

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: Western Blot, In Vitro, Electrophoresis, Control, Purification, Protein Concentration, Software

The representative HPLC traces of GFP-tagged PCSK9 synthetic protein after addition of PNA oligomers ( A ). Area under the curve (AUC) values were calculated for each sample (run in triplicates) from chromatography peaks of GFP-tagged PCSK9 protein synthesized with the presence of different PNA oligomers ( B ) *: p < 0.05 vs. CtrlPNA.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: The representative HPLC traces of GFP-tagged PCSK9 synthetic protein after addition of PNA oligomers ( A ). Area under the curve (AUC) values were calculated for each sample (run in triplicates) from chromatography peaks of GFP-tagged PCSK9 protein synthesized with the presence of different PNA oligomers ( B ) *: p < 0.05 vs. CtrlPNA.

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: Chromatography, Synthesized

Sequences of synthesized PNA-Ahx-NLS conjugates (Ahx-6-aminohexanoic acid).

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Sequences of synthesized PNA-Ahx-NLS conjugates (Ahx-6-aminohexanoic acid).

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: Synthesized, Sequencing

Properties of PNA oligomers used in this study.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Properties of PNA oligomers used in this study.

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: Sequencing, Control

High-performance liquid chromatography (HPLC) conditions for analysis of GFP-tagged PCSK9 protein after cell-free synthesis in vitro. The reaction was carried out under the following conditions: flow: 0.8 mL/min; detection (ex/em): 482/511 nm; column temperature: 40 °C; injection volume: 10 µL; mobile phase A: 0.1% trifluoroacetic acid in deionized water; mobile phase B: 0.1% trifluoroacetic acid in acetonitrile.

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: High-performance liquid chromatography (HPLC) conditions for analysis of GFP-tagged PCSK9 protein after cell-free synthesis in vitro. The reaction was carried out under the following conditions: flow: 0.8 mL/min; detection (ex/em): 482/511 nm; column temperature: 40 °C; injection volume: 10 µL; mobile phase A: 0.1% trifluoroacetic acid in deionized water; mobile phase B: 0.1% trifluoroacetic acid in acetonitrile.

Article Snippet: A pCMV6-AC-GFP plasmid coding for the PCSK9 gene was purchased from OriGene Technologies (Rockville, MD, USA, Cat No. RG220000).

Techniques: High Performance Liquid Chromatography, In Vitro, Injection

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques:

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Reverse Transcription, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Sequencing, Real-time Polymerase Chain Reaction, Comparison

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Western Blot, In Vitro, Electrophoresis, Control, Purification, Protein Concentration, Software

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Chromatography, Synthesized

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Sequences of synthesized PNA-Ahx-NLS conjugates (Ahx-6-aminohexanoic acid).

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Synthesized, Sequencing

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Figure Lengend Snippet: Properties of PNA oligomers used in this study.

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: Sequencing, Control

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms25031463

Article Snippet: A circular pCMV6-AC-GFP plasmid containing the coding sequence for PCSK9 (OriGene Technologies, Rockville, MD, USA, Cat No. RG220000) served as a template.

Techniques: High Performance Liquid Chromatography, In Vitro, Injection

Main clinical characteristics of patients and results of genetic testing.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Main clinical characteristics of patients and results of genetic testing.

Article Snippet: The LDLR -3′ UTR construct ( LDLR NM_00052 human 3′UTR clone, WT) and the PCSK9 -3′ UTR construct ( PCSK9 NM_174936 human 3'UTR clone, WT) were obtained from OriGene to perform the luciferase reporter experiment assays.

Techniques:

Clinical and biochemical characteristics of patients with LDLR and PCSK9 -3′UTR variants.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Clinical and biochemical characteristics of patients with LDLR and PCSK9 -3′UTR variants.

Techniques:

Schematic representation of the 3′UTR variants found in the patients: (a) LDLR -3′UTR variants; (b) PCSK9 -3′UTR variants.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Schematic representation of the 3′UTR variants found in the patients: (a) LDLR -3′UTR variants; (b) PCSK9 -3′UTR variants.

Techniques:

Comparison of the luciferase activity of 3′UTR-PCSK9 variants using the luciferase reporter assay. The 3′UTR-PCSK9 luciferase reporter construct WT (WT) or mutants c. ∗ 171C > T (c.171), c. ∗ 234C > T (c.234), and c. ∗ 950C > T (c.950) were transfected in HepG2 cells. The luciferase activity of the cell lysates was then measured by luminescence. Each result corresponds to the mean ± standard deviation of the triplicate assays of the luciferase activity of the cell lysates of 3′UTR-PCSK9 luciferase reporter construct WT (WT) or mutants: c. ∗ 171C > T (c.171), c. ∗ 234C > T (c.234), and c. ∗ 950C > T (c.950). Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained from three independent experiments.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Comparison of the luciferase activity of 3′UTR-PCSK9 variants using the luciferase reporter assay. The 3′UTR-PCSK9 luciferase reporter construct WT (WT) or mutants c. ∗ 171C > T (c.171), c. ∗ 234C > T (c.234), and c. ∗ 950C > T (c.950) were transfected in HepG2 cells. The luciferase activity of the cell lysates was then measured by luminescence. Each result corresponds to the mean ± standard deviation of the triplicate assays of the luciferase activity of the cell lysates of 3′UTR-PCSK9 luciferase reporter construct WT (WT) or mutants: c. ∗ 171C > T (c.171), c. ∗ 234C > T (c.234), and c. ∗ 950C > T (c.950). Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained from three independent experiments.

Techniques: Comparison, Luciferase, Activity Assay, Reporter Assay, Construct, Transfection, Standard Deviation

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Schematic representation of the 3′UTR variants found in the patients: (a) LDLR -3′UTR variants; (b) PCSK9 -3′UTR variants.

Techniques:

Comparison of the luciferase activity of LDLR-3′UTR variants using luciferase reporter assay. LDLR-3′UTR WT luciferase reporter construct or mutant-plasmids c. ∗ 19G > A, c. ∗ 503C > T, c. ∗ 517C > A, c. ∗ 653G > C, and c. ∗ 1227C > T were transfected in HepG2 cells. The luciferase activity of cell lysates was then measured by chemiluminescence. Each result corresponds to the mean ± standard deviation of the triplicate assays of the luciferase activity of the LDLR-3′UTR WT luciferase reporter construct or mutant-plasmids: c. ∗ 19G > A (c ∗ 19), c. ∗ 503C > T (c ∗ 503), c. ∗ 517C > A (c ∗ 517), c. ∗ 653G > C (c ∗ 653), and c. ∗ 1227C > T (c ∗ 1227). Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Comparison of the luciferase activity of LDLR-3′UTR variants using luciferase reporter assay. LDLR-3′UTR WT luciferase reporter construct or mutant-plasmids c. ∗ 19G > A, c. ∗ 503C > T, c. ∗ 517C > A, c. ∗ 653G > C, and c. ∗ 1227C > T were transfected in HepG2 cells. The luciferase activity of cell lysates was then measured by chemiluminescence. Each result corresponds to the mean ± standard deviation of the triplicate assays of the luciferase activity of the LDLR-3′UTR WT luciferase reporter construct or mutant-plasmids: c. ∗ 19G > A (c ∗ 19), c. ∗ 503C > T (c ∗ 503), c. ∗ 517C > A (c ∗ 517), c. ∗ 653G > C (c ∗ 653), and c. ∗ 1227C > T (c ∗ 1227). Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Techniques: Comparison, Luciferase, Activity Assay, Reporter Assay, Construct, Mutagenesis, Transfection, Standard Deviation

Effect of miR-296-3p on the expression of LDLR -3′UTR variants (a) c. ∗ 19G > A, (b) c. ∗ 503C > T, (c) c. ∗ 517C > A, and (d) c. ∗ 1227C > T in the luciferase reporter assay. 3′UTR- LDLR WT (WT) luciferase reporter construct or mutant-plasmids c. ∗ 19G > A, c. ∗ 503C > T, c. ∗ 517C > A, c. ∗ 653G > C, and c. ∗ 1227C > T were cotransfected with miR-296-3p mimic or mimic negative control; the luciferase activity of the cell lysates was then measured. The luciferase activity of the variants cotransfected with miR-296-3p mimic or mimic negative control (CN) is shown in (a) c. ∗ 19G > A (c ∗ 19), (b) c. ∗ 503C > T (c ∗ 503), (c) c. ∗ 517C > A (c ∗ 517), and (d) c. ∗ 1227C > T (c ∗ 1227). Each result corresponds to the percentage of luciferase activity with respect to LDLR -3′UTR WT with mimic negative control ± standard deviation of the triplicate assays. Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Effect of miR-296-3p on the expression of LDLR -3′UTR variants (a) c. ∗ 19G > A, (b) c. ∗ 503C > T, (c) c. ∗ 517C > A, and (d) c. ∗ 1227C > T in the luciferase reporter assay. 3′UTR- LDLR WT (WT) luciferase reporter construct or mutant-plasmids c. ∗ 19G > A, c. ∗ 503C > T, c. ∗ 517C > A, c. ∗ 653G > C, and c. ∗ 1227C > T were cotransfected with miR-296-3p mimic or mimic negative control; the luciferase activity of the cell lysates was then measured. The luciferase activity of the variants cotransfected with miR-296-3p mimic or mimic negative control (CN) is shown in (a) c. ∗ 19G > A (c ∗ 19), (b) c. ∗ 503C > T (c ∗ 503), (c) c. ∗ 517C > A (c ∗ 517), and (d) c. ∗ 1227C > T (c ∗ 1227). Each result corresponds to the percentage of luciferase activity with respect to LDLR -3′UTR WT with mimic negative control ± standard deviation of the triplicate assays. Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Techniques: Expressing, Luciferase, Reporter Assay, Construct, Mutagenesis, Negative Control, Activity Assay, Standard Deviation

Effect of miR-449c-5p on the expression of LDLR -3′UTR variant c. ∗ 653G > C in the luciferase reporter assay. The LDLR -3′UTR luciferase reporter construct WT (WT) and mutated-plasmid LDLR :c. ∗ 653G > C (c ∗ 653) were cotransfected with miR-449c-5p mimic or negative mimic control (CN). The luciferase activity of the cell lysates was then measured. The luciferase activity of the WT and mutant c. ∗ 653G > C cotransfected with miR449c-5p or mimic negative control is shown. Each result corresponds to the percentage of luciferase activity with respect to WT with mimic negative control ± standard deviation of the triplicate assays. Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Figure Lengend Snippet: Effect of miR-449c-5p on the expression of LDLR -3′UTR variant c. ∗ 653G > C in the luciferase reporter assay. The LDLR -3′UTR luciferase reporter construct WT (WT) and mutated-plasmid LDLR :c. ∗ 653G > C (c ∗ 653) were cotransfected with miR-449c-5p mimic or negative mimic control (CN). The luciferase activity of the cell lysates was then measured. The luciferase activity of the WT and mutant c. ∗ 653G > C cotransfected with miR449c-5p or mimic negative control is shown. Each result corresponds to the percentage of luciferase activity with respect to WT with mimic negative control ± standard deviation of the triplicate assays. Statistical significance was determined by a Welch's correction unpaired t -test (2-sided) and a 95% confidence interval. The results were obtained in three independent experiments.

Techniques: Expressing, Variant Assay, Luciferase, Reporter Assay, Construct, Plasmid Preparation, Control, Activity Assay, Mutagenesis, Negative Control, Standard Deviation

Journal: Human Mutation

Article Title: Functional Analysis of 3′UTR Variants at the LDLR and PCSK9 Genes in Patients with Familial Hypercholesterolemia

doi: 10.1155/2024/9964734

Techniques: Comparison, Luciferase, Activity Assay, Reporter Assay, Construct, Transfection, Standard Deviation

Figure 1. High PCSK9 expression is associated with poor prognosis in patients with ESCC. (A) Immunohistochemistry of normal esophageal epithelial and ESCC tissues from patients with ESCC. Typical cases with PCSK9 expression are shown. Scale bar, 100 µm. (B) PCSK9 expression levels in an ESCC validation set obtained from The Cancer Genome Atlas database. (C) Differential expression of PCSK9 in ESCC tissues and normal esophageal epithelial tissues from 60 patients. (D) PCSK9 expression levels in freshly obtained esophageal cancer tissues compared with adjacent paracancerous tissues in five patients. (E) Based on the expression of PCSK9, Kaplan‑Meier analysis showed that high PCSK9 was associated with a shorter overall survival in 100 patients with ESCC. (F) In the PCSK9‑high expression group, the clinical stage was higher than that in the PCSK9‑low expression group. (G) PCSK9‑high expression groups demonstrated a higher rate of lymph node metastasis. (H) A higher T classification was observed in the PCSK9‑high expression group than that in the PCSK9‑low expression group. Scale bars, 100 µm. *P<0.05, **P<0.01, ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; N, normal; T, tumor.

Journal: Oncology letters

Article Title: PCSK9 promotes tumor cell proliferation and migration by facilitating CCL25 secretion in esophageal squamous cell carcinoma.

doi: 10.3892/ol.2023.14086

Figure Lengend Snippet: Figure 1. High PCSK9 expression is associated with poor prognosis in patients with ESCC. (A) Immunohistochemistry of normal esophageal epithelial and ESCC tissues from patients with ESCC. Typical cases with PCSK9 expression are shown. Scale bar, 100 µm. (B) PCSK9 expression levels in an ESCC validation set obtained from The Cancer Genome Atlas database. (C) Differential expression of PCSK9 in ESCC tissues and normal esophageal epithelial tissues from 60 patients. (D) PCSK9 expression levels in freshly obtained esophageal cancer tissues compared with adjacent paracancerous tissues in five patients. (E) Based on the expression of PCSK9, Kaplan‑Meier analysis showed that high PCSK9 was associated with a shorter overall survival in 100 patients with ESCC. (F) In the PCSK9‑high expression group, the clinical stage was higher than that in the PCSK9‑low expression group. (G) PCSK9‑high expression groups demonstrated a higher rate of lymph node metastasis. (H) A higher T classification was observed in the PCSK9‑high expression group than that in the PCSK9‑low expression group. Scale bars, 100 µm. *P<0.05, **P<0.01, ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; N, normal; T, tumor.

Article Snippet: The PCSK9 plasmid (cat. no. RC220000; Origene Technologies, Inc.) was employed to induce overex‐ pression of PCSK9, whereas the empty pCMV6‐Entry plasmid (cat. no. PS100001; Origene Technologies, Inc.) served as a control.

Techniques: Expressing, Immunohistochemistry, Biomarker Discovery, Quantitative Proteomics

Figure 2. PCSK9 knockdown and overexpression in ESCC cell lines. (A) In ESCC cell lines, the mRNA expression levels of PCSK9 were lowest in TE1 cells and highest in KYSE150 cells. (B) Transfection efficiency of si‑PCSK9 in the KYSE150 cell line; subsequent experiments were conducted using si‑PCSK9‑2, which had the highest knockdown efficiency. (C) Transfection efficiency of PCSK9 overexpression in the TE1 cell line. (D) PCSK9 protein expression following PCSK9 knockdown in KYSE150 cells. (E) PCSK9 protein expression following PCSK9 overexpression in TE1 cells. *P<0.05, **P<0.01. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; si, small interfering; NC, negative control.

Journal: Oncology letters

Article Title: PCSK9 promotes tumor cell proliferation and migration by facilitating CCL25 secretion in esophageal squamous cell carcinoma.

doi: 10.3892/ol.2023.14086

Figure Lengend Snippet: Figure 2. PCSK9 knockdown and overexpression in ESCC cell lines. (A) In ESCC cell lines, the mRNA expression levels of PCSK9 were lowest in TE1 cells and highest in KYSE150 cells. (B) Transfection efficiency of si‑PCSK9 in the KYSE150 cell line; subsequent experiments were conducted using si‑PCSK9‑2, which had the highest knockdown efficiency. (C) Transfection efficiency of PCSK9 overexpression in the TE1 cell line. (D) PCSK9 protein expression following PCSK9 knockdown in KYSE150 cells. (E) PCSK9 protein expression following PCSK9 overexpression in TE1 cells. *P<0.05, **P<0.01. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; si, small interfering; NC, negative control.

Techniques: Knockdown, Over Expression, Expressing, Transfection, Negative Control

Figure 3. PCSK9 promotes cell proliferation, migration and invasion in vitro. (A) Proliferation of TE1 cells overexpressing PCSK9 and those transfected with a vector control, as determined using CCK‑8 assay. PCSK9 promoted ESCC cell proliferation. ***P<0.001 vs. Control. (B) Colony formation assay of TE1 cells; PCSK9 overexpression promoted colony formation. Migration and invasion of TE1 cells with PCSK9 overexpression were detected using (C) Transwell migration and invasion assays, and (D) wound healing assays. PCSK9 promoted the invasion and migration of ESCC cells. (E) Proliferation of KYSE150 cells with PCSK9 knockdown and those transfected with si‑NC, as determined using CCK‑8 assay. (F) Colony formation assay of KYSE150 cells. (G) Transwell migration and invasion, and (H) wound healing assays detected migration and invasion of KYSE150 cells. Scale bars, 100 µm. **P<0.01. ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; CCK‑8, Cell Counting Kit‑8; ESCC, esophageal squamous cell carcinoma; si, small interfering; NC, negative control.

Journal: Oncology letters

Article Title: PCSK9 promotes tumor cell proliferation and migration by facilitating CCL25 secretion in esophageal squamous cell carcinoma.

doi: 10.3892/ol.2023.14086

Figure Lengend Snippet: Figure 3. PCSK9 promotes cell proliferation, migration and invasion in vitro. (A) Proliferation of TE1 cells overexpressing PCSK9 and those transfected with a vector control, as determined using CCK‑8 assay. PCSK9 promoted ESCC cell proliferation. ***P<0.001 vs. Control. (B) Colony formation assay of TE1 cells; PCSK9 overexpression promoted colony formation. Migration and invasion of TE1 cells with PCSK9 overexpression were detected using (C) Transwell migration and invasion assays, and (D) wound healing assays. PCSK9 promoted the invasion and migration of ESCC cells. (E) Proliferation of KYSE150 cells with PCSK9 knockdown and those transfected with si‑NC, as determined using CCK‑8 assay. (F) Colony formation assay of KYSE150 cells. (G) Transwell migration and invasion, and (H) wound healing assays detected migration and invasion of KYSE150 cells. Scale bars, 100 µm. **P<0.01. ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; CCK‑8, Cell Counting Kit‑8; ESCC, esophageal squamous cell carcinoma; si, small interfering; NC, negative control.

Techniques: Migration, In Vitro, Transfection, Plasmid Preparation, Control, CCK-8 Assay, Colony Assay, Over Expression, Knockdown, Negative Control

Figure 4. PCSK9 modulates the EMT of ESCC cells. (A) PCSK9 regulates esophageal squamous cell carcinoma progression and metastasis in TE1 cells through EMT. Overexpression of PCSK9 inhibits E‑cadherin expression, and upregulates N‑cadherin and vimentin protein expression. (B) Western blotting revealed that PCSK9 knockdown in the KYSE150 cells increased E‑cadherin, and inhibited N‑cadherin and vimentin expression. *P<0.05, **P<0.01. PCSK9, proprotein convertase subtilisin/kexin type 9; si, small interfering; NC, negative control; EMT, epithelial‑mesenchymal transition.

Journal: Oncology letters

Article Title: PCSK9 promotes tumor cell proliferation and migration by facilitating CCL25 secretion in esophageal squamous cell carcinoma.

doi: 10.3892/ol.2023.14086

Figure Lengend Snippet: Figure 4. PCSK9 modulates the EMT of ESCC cells. (A) PCSK9 regulates esophageal squamous cell carcinoma progression and metastasis in TE1 cells through EMT. Overexpression of PCSK9 inhibits E‑cadherin expression, and upregulates N‑cadherin and vimentin protein expression. (B) Western blotting revealed that PCSK9 knockdown in the KYSE150 cells increased E‑cadherin, and inhibited N‑cadherin and vimentin expression. *P<0.05, **P<0.01. PCSK9, proprotein convertase subtilisin/kexin type 9; si, small interfering; NC, negative control; EMT, epithelial‑mesenchymal transition.

Techniques: Over Expression, Expressing, Western Blot, Knockdown, Negative Control

Figure 5. PCSK9 promotes ESCC progression through CCL25 upregulation. (A) PCSK9 data were extracted from The Cancer Genome Atlas public database and divided into high and low expression groups, according to the expression of the PCSK9. The original count matrix of the selected public data was analyzed for differences according to the standard process using the DESeq2 package. CCL25 expression was also low in the PCSK9‑low expression group. (B) As determined by ELISA, PCSK9 overexpression increased CCL25 secretion, whereas PCSK9 knockdown inhibited CCL25 secretion. (C) Cell Counting Kit‑8 assay showed that the exogenous addition of CCL25 inhibited PCSK9 knockdown‑induced inhibition of cell proliferation. (D) Colony formation assay showed that the exogenous addition of CCL25 rescued PCSK9 knockdown‑induced colony formation inhibition. (E) Transwell migration and invasion, and (F) wound healing assays were performed to examine whether the exogenous addition of CCL25 could reverse PCSK9 knockdown‑induced inhibition of the migration and invasion of ESCC cells. Scale bars, 100 µm. **P<0.01, ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; CCL25, chemokine (C‑C motif) ligand 25; si, small interfering; NC, negative control.

Journal: Oncology letters

Article Title: PCSK9 promotes tumor cell proliferation and migration by facilitating CCL25 secretion in esophageal squamous cell carcinoma.

doi: 10.3892/ol.2023.14086

Figure Lengend Snippet: Figure 5. PCSK9 promotes ESCC progression through CCL25 upregulation. (A) PCSK9 data were extracted from The Cancer Genome Atlas public database and divided into high and low expression groups, according to the expression of the PCSK9. The original count matrix of the selected public data was analyzed for differences according to the standard process using the DESeq2 package. CCL25 expression was also low in the PCSK9‑low expression group. (B) As determined by ELISA, PCSK9 overexpression increased CCL25 secretion, whereas PCSK9 knockdown inhibited CCL25 secretion. (C) Cell Counting Kit‑8 assay showed that the exogenous addition of CCL25 inhibited PCSK9 knockdown‑induced inhibition of cell proliferation. (D) Colony formation assay showed that the exogenous addition of CCL25 rescued PCSK9 knockdown‑induced colony formation inhibition. (E) Transwell migration and invasion, and (F) wound healing assays were performed to examine whether the exogenous addition of CCL25 could reverse PCSK9 knockdown‑induced inhibition of the migration and invasion of ESCC cells. Scale bars, 100 µm. **P<0.01, ***P<0.001. PCSK9, proprotein convertase subtilisin/kexin type 9; ESCC, esophageal squamous cell carcinoma; CCL25, chemokine (C‑C motif) ligand 25; si, small interfering; NC, negative control.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Over Expression, Knockdown, CCK-8 Assay, Inhibition, Colony Assay, Migration, Negative Control

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