Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 is overexpressed in OSCC and is associated with a poor prognosis. Relative mRNA expression of CMKLR1 in (A) HNSCC tissues ( n = 520) versus adjacent normal tissues ( n = 44) and in (B) non–lymph node (N0) versus lymph node (N1) samples, according to TCGA database analysis. (C) Kaplan–Meier survival analysis of OS based on CMKLR1 expression in patients with HNSCC, with data from TCGA. (D) Representative IHC images depicting CMKLR1 expression in OSCC tissues and adjacent normal oral mucosal tissues ( n = 68). Scale bar = 20 μm. (E) Quantitative analysis of CMKLR1 IHC staining intensity in OSCC samples ( n = 68) versus normal tissue samples, with subgroup comparisons based on T stage, clinical stage, and lymph node metastasis. (F) Kaplan–Meier survival analysis of patients with OSCC with high versus low CMKLR1 expression according to IHC scores. (G) RT-qPCR analysis of CMKLR1 mRNA expression in 50 paired OSCC and adjacent normal tissues. (H) Western blot analysis of CMKLR1 protein levels in five representative paired OSCC (T) and adjacent nontumorous (N) tissues. N, adjacent nontumorous tissues; T, OSCC tumor tissues; TCGA, The Cancer Genome Atlas. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Expressing, Immunohistochemistry, Quantitative RT-PCR, Western Blot

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes OSCC cell proliferation and tumor growth in vivo. (A) Western blot analysis confirming CMKLR1 knockdown in Cal-27 cells (Sh1 and Sh2) and CMKLR1 overexpression in SCC-9 cells (OE). (B) CCK-8 proliferation assays of the proliferation of CMKLR1-silenced Cal-27 cells and CMKLR1-overexpressing SCC-9 cells. (C, D) Colony formation assays and quantification in the indicated groups. (E) Representative images of xenograft tumors derived from OSCC cells with CMKLR1 knockdown (Sh), OSCC cells with CMKLR1 overexpression (OE), and control cells (NC). (F) Tumor growth curves demonstrating tumor volume changes over time. (G) Tumor weight measurements at the endpoint. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: In Vivo, Western Blot, Knockdown, Over Expression, CCK-8 Assay, Derivative Assay, Control

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes OSCC cell migration, invasion, and EMT. Representative images and quantification of wound healing assays in (A) Cal-27 cells with CMKLR1 knockdown and (B) SCC-9 cells with CMKLR1 overexpression. Transwell invasion assays and quantification in (C) Cal-27 and (D) SCC-9 cells, revealing a decreased and increased invasive capacity after CMKLR1 knockdown and overexpression, respectively. Scale bar = 100 μm. (E) Western blot analysis of EMT markers (E-cadherin, N-cadherin, vimentin, Snail, and Slug) in CMKLR1-modified OSCC cells. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Migration, Knockdown, Over Expression, Western Blot, Modification

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 knockdown suppresses mitochondrial OXPHOS in OSCC cells. (A) Volcano plot of DEPs in CMKLR1-knockdown and control Cal-27 cells, depicting upregulated proteins (red), downregulated proteins (blue), and nonsignificant proteins (gray). (B) Heatmap of the top DEPs identified through proteomic analysis. (C) Subcellular localization pie chart of DEPs, revealing a large proportion localized to mitochondria. (D, E) GO and KEGG pathway enrichment analyses of DEPs, highlighting enrichment in OXPHOS-related processes. (F) Gene set enrichment analysis demonstrating the significant downregulation of OXPHOS in CMKLR1-silenced cells. (G) Seahorse XF analysis of mitochondrial respiration indicating a reduced OCR in CMKLR1-knockdown OSCC cells. (H) Quantification of basal respiration, ATP production, maximal respiration, and spare respiratory capacity in CMKLR1-knockdown cells. (I) Western blot analysis of representative OXPHOS complex subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC1, CIV-MTCO2, and CV-ATP5A1), with β-actin as loading control. (J) Representative MitoTracker images and quantification of fluorescence intensity (MitoTracker Red, scale bar = 20 μm). * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Knockdown, Control, Western Blot, Fluorescence

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: CMKLR1 promotes mitochondrial OXPHOS through the PI3K/AKT/PGC-1α axis. (A) Immunoblot analysis of p-PI3K, PI3K, p-AKT, AKT, and PGC-1α in OSCC cells with CMKLR1 knockdown. (B) Protein levels of p-PI3K, p-AKT, and PGC-1α were measured in OSCC cells overexpressing CMKLR1, with or without LY294002 treatment (10 μM). (C) Western blot analysis of representative OXPHOS complex subunits (CI-NDUFB8, CII-SDHB, CIII-UQCRC1, CIV-MTCO2, and CV-ATP5A1) in CMKLR1-overexpressing cells after transfection with siPGC-1α or siNC. (D) Seahorse XF analysis of OCR curves. (E) Quantification of basal respiration, ATP-linked respiration, maximal respiration, and spare respiratory capacity. (F) Representative MitoTracker images and quantification of fluorescence intensity (MitoTracker Red, scale bar = 20 μm). * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Western Blot, Knockdown, Transfection, Fluorescence

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: Inhibition of OXPHOS reverses the impact of CMKLR1-mediated tumor-promoting effects. (A) CCK-8 assay of the proliferation of CMKLR1-overexpressing (OE) OSCC cells treated with IACS-010759. (B) Colony formation assay of NC, OE, and OE + IACS-010759 cells. The right panel presents the number of colonies. (C, D) Transwell assay revealing significantly impaired cell invasion and migration after OXPHOS inhibitor treatment in CMKLR1-overexpressing cells. Scale bar = 100 μm. (E) Representative images of xenograft tumors derived from NC, OE, and OE + IACS-010759 cells. (F) Tumor growth curves demonstrating tumor volume changes over time. (G) Tumor weight measurements at the endpoint. (H) Representative images of IHC staining of tumor sections with Ki-67. Scale bar = 50 μm. (I) Quantification of Ki67 expression in xenograft tumors. * P < .05; ** P < .01; *** P < .001.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: Inhibition, CCK-8 Assay, Colony Assay, Transwell Assay, Migration, Derivative Assay, Immunohistochemistry, Expressing

Journal: International Dental Journal

Article Title: Chemokine-Like Receptor 1 Knockdown Suppresses Oral Squamous Cell Carcinoma Progression by Reducing Oxidative Phosphorylation

doi: 10.1016/j.identj.2026.109479

Figure Lengend Snippet: Schematic diagram of the mechanism by which CMKLR1 facilitates OSCC progression.

Article Snippet: Short hairpin RNA (shRNA) lentiviral vectors targeting human CMKLR1 were purchased from OBiO Technology (Shanghai, China).

Techniques: