Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: The correlation of P. gingivalis abundance and cancer stem cell markers expression in OSCC tissues. a Representative IHC images of ALDH1, BMI1, NANOG, and SOX2 in OSCC samples. Scale bar: 50 μm. b Quantitative analysis of ALDH1, BMI1, NANOG, and SOX2 in OSCC samples. n = 30. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01. c The correlations between P. gingivalis and the expression of ALDH1, BMI1, NANOG, and SOX2 were analyzed with Pearson correlation analysis. n = 20

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Expressing

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: Persistent exposure to P. gingivalis promoted OSCC cells to acquire stem-like features. a Colony formation showed P. gingivalis increased the number of cell clones. b Sphere formation showed P. gingivalis increased the diameter of tumorspheres. Scale bar: 100 μm. c , d Western blot and quantification showed the upregulation of stemness signature proteins BMI1, NANOG, and SOX2 compared to the control group. β-actin was used as a housekeeping gene. e Flow cytometry showed P. gingivalis increased the proportion of ALDH1 + subpopulation of HSC-4 and SCC-9 cells. n = 3. Data are presented as mean ± SD. ns: P > 0.05, * P < 0.05, ** P < 0.01

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Clone Assay, Western Blot, Control, Flow Cytometry

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: P. gingivalis induced OSCC cells to acquire stem-like features by regulating lipid synthesis. a Volcano plot of differentially expressed genes (DEGs) between HSC-4 cells treated with and without P. gingivalis . b Top twenty most enriched GO (BP) pathways enrichment analysis for differential genes were visible. c Nile red staining showed Triacsin C abolished P. gingivalis -induced lipid droplets accumulation. Scale bar: 20 μm. d Sphere formation showed Triacsin C decreased P. gingivalis -induced the diameter of tumorspheres. Scale bar: 100 μm. e Western blot showed Triacsin C significantly suppressed P. gingivalis -induced upregulation of BMI1, NANOG, and SOX2. β-actin was used as a housekeeping gene. n = 3

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Staining, Western Blot

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: P. gingivalis induced OSCC cells to acquire stem-like features by modulating SCD1-mediated lipid synthesis. a , b Colony formation and quantification showed knockdown of SCD1 decreased P. gingivalis -induced the number of cell clones. c , d Sphere formation and quantification showed knockdown of SCD1 decreased P. gingivalis -induced the number and diameter of tumorspheres. Scale bar: 100 μm. n = 3. Data are presented as mean ± SD. ** P < 0.01

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Knockdown, Clone Assay

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: P. gingivalis induced OSCC cells to express stemness markers by modulating SCD1-mediated lipid synthesis. a , b Western blot and quantification showed knockdown of SCD1 suppressed P. gingivalis -induced upregulation of BMI1, NANOG, and SOX2. β-actin was used as a housekeeping gene. c , d Flow cytometry and quantification showed knockdown of SCD1 decreased P. gingivalis- increased the proportion of ALDH1 + subpopulation of OSCC cells. n = 3. Data are presented as mean ± SD. * P < 0.05, ** P < 0.01

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Western Blot, Knockdown, Flow Cytometry

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: SCD1 upregulation in OSCC cells was transcriptionally activated by KLF5. a Venn diagram showing the candidate transcription factor screening process. b , c qRT-PCR showed P. gingivalis specifically promoted the expression of KLF5. d ChIP-qPCR showed the binding of KLF5 to the SCD1 promoter region in HSC-4 cells. e Dual-luciferase reporter assay showed the binding of KLF5 to the SCD1 promoter region in 293 T cells. f Western blot showed knockdown of KLF5 suppressed the protein level of SCD1. β-actin was used as a housekeeping gene. n = 3. Data are presented as the mean ± SD. ns: P > 0.05, * P < 0.05, ** P < 0.01

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Quantitative RT-PCR, Expressing, ChIP-qPCR, Binding Assay, Luciferase, Reporter Assay, Western Blot, Knockdown

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: P. gingivalis stabilized KLF5 expression via the activation of NOD1. a Top twenty most enriched KEGG pathways among significantly upregulated DEGs in HSC-4 cells co-cultured with P. gingivalis . b qRT-PCR showed P. gingivalis promoted the NOD1 gene expression of OSCC cells. c – f Western blot and quantification showed silencing of NOD1 decreased the expression of KLF5 in OSCC cells. β-actin was used as a housekeeping gene. n = 3. Data are presented as the mean ± SD. ns: P > 0.05, * P < 0.05, ** P < 0.01

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Expressing, Activation Assay, Cell Culture, Quantitative RT-PCR, Gene Expression, Western Blot

Journal: International Journal of Oral Science

Article Title: Porphyromonas gingivalis potentiates stem-like properties of oral squamous cell carcinoma by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis

doi: 10.1038/s41368-024-00342-8

Figure Lengend Snippet: Schematic drawing of the role of P. gingivalis in regulating OSCC stemness. The specific molecular mechanism is that P. gingivalis promotes OSCC stemness by modulating SCD1-dependent lipid synthesis via NOD1/KLF5 axis. The figure was drawn using Figdraw

Article Snippet: The OSCC cell lines HSC-4 and SCC-9 were originally derived from a patient with tongue cancer and respectively obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB; Shinjuku, Japan) and American Type Culture Collection (ATCC, Manassas, USA).

Techniques:

Journal: Oncology Letters

Article Title: Knockdown of TFAP2E results in rapid G 2 /M transition in oral squamous cell carcinoma cells

doi: 10.3892/ol.2024.14260

Figure Lengend Snippet: siRNA-mediated knockdown of TFAP2E accelerates OSCC cell proliferation. OSCC-derived Ca9-22 and HSC-4 cells were transfected with si-TFAP2E or si-N/C. A total of 2 days after transfection, total RNA and cell lysates were prepared and analyzed by (A) reverse transcription-quantitative PCR and (B) western blotting, respectively. Data are presented as the mean ± SD of triplicate measurements. *P<0.05, **P<0.01. GAPDH was used as a loading control for western blotting. Proliferation of (C) Ca9-22 and (E) HSC-4 cells was determined by water-soluble tetrazolium salt-8 assay at the indicated time points post-transfection. Data are presented as the mean ± SD of quadruplicate measurements. **P<0.01 vs. si-N/C. To visualize viable cells, cells were fixed and stained using a Diff-Quick Stain Kit 10 days after transfection. Representative images of (D) Ca9-22 and (F) HSC-4 are shown. N/C, negative control; siRNA, small interfering RNA.

Article Snippet: The human gingival cancer cell line Ca9-22 was obtained from the Japanese Collection of Research Bioresources Cell Bank and the human tongue cancer-derived cell line HSC-4 was obtained from the RIKEN BioResource Center.

Techniques: Derivative Assay, Transfection, Real-time Polymerase Chain Reaction, Western Blot, Staining, Diff-Quik, Negative Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: Knockdown of TFAP2E results in rapid G 2 /M transition in oral squamous cell carcinoma cells

doi: 10.3892/ol.2024.14260

Figure Lengend Snippet: TFAP2E-knockdown has no detectable effect on cell cycle distribution patterns. (A and C) Ca9-22 and (B and D) HSC-4 cells were transfected with siRNA-TFAP2E or siRNA-N/C. A total of 3 days after transfection, floating and adherent cells were harvested and stained with propidium iodide, and their cell cycle distributions were examined using fluorescence-activated cell sorting. The experiments were performed at least three times, and the percentage distribution was calculated based on Michael H. Fox algorithm. (A and B) Representative histograms are shown. (C and D) Data are presented as the mean ± SD of triplicate measurements. Sub-G 1 phase indicates the cell population, in which DNA content was lower than that in the G 0 /G 1 phase. N/C, negative control; siRNA, small interfering RNA.

Article Snippet: The human gingival cancer cell line Ca9-22 was obtained from the Japanese Collection of Research Bioresources Cell Bank and the human tongue cancer-derived cell line HSC-4 was obtained from the RIKEN BioResource Center.

Techniques: Transfection, Staining, Fluorescence, FACS, Negative Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: Knockdown of TFAP2E results in rapid G 2 /M transition in oral squamous cell carcinoma cells

doi: 10.3892/ol.2024.14260

Figure Lengend Snippet: Knockdown of TFAP2E causes an earlier entry into M phase. (A and C) Ca9-22 and (B and D) HSC-4 cells were transfected with siRNA-TFAP2E or siRNA-N/C. A total of 2 days after transfection, the culture medium was replaced with fresh medium containing nocodazole. Both floating and adherent cells were harvested at the indicated time points and subjected to fluorescence-activated cell sorting analysis. (A and B) Percentage distribution was calculated based on Michael H. Fox algorithm. Data are presented as the means ± SD of triplicate measurements. (C and D) Percentage of p-H3Ser28-positive cells. Data are presented as the mean ± SD of triplicate measurements. *P<0.05, **P<0.01 as indicated or vs. siRNA-N/C. N/C, negative control; siRNA, small interfering RNA.

Article Snippet: The human gingival cancer cell line Ca9-22 was obtained from the Japanese Collection of Research Bioresources Cell Bank and the human tongue cancer-derived cell line HSC-4 was obtained from the RIKEN BioResource Center.

Techniques: Transfection, Fluorescence, FACS, Negative Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: Knockdown of TFAP2E results in rapid G 2 /M transition in oral squamous cell carcinoma cells

doi: 10.3892/ol.2024.14260

Figure Lengend Snippet: Accumulation of p-H3Ser28 and cyclin B1 is promoted in oral squamous cell carcinoma cells after TFAP2E knockdown compared with in the control group. (A) Ca9-22 and (C) HSC-4 cells were transfected with siRNA-TFAP2E or siRNA-N/C, and were treated with nocodazole 2 days after the transfection. Both floating and adherent cells were harvested at the indicated time points, and whole-cell lysates were subjected to western blotting analysis. GAPDH was used as a loading control. The band density of p-H3Ser28 was normalized to that of total H3 in (B) Ca9-22 and (D) HSC-4 cells. Data are presented as the mean ± SD of triplicate measurements. *P<0.05 vs. siRNA-N/C. N/C, negative control; p-H3Ser28, phosphorylated histone H3 serine 28; siRNA, small interfering RNA.

Article Snippet: The human gingival cancer cell line Ca9-22 was obtained from the Japanese Collection of Research Bioresources Cell Bank and the human tongue cancer-derived cell line HSC-4 was obtained from the RIKEN BioResource Center.

Techniques: Transfection, Western Blot, Negative Control, Small Interfering RNA