Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: Construction and validation of the RRLD model in vitro. a Study protocol for the construction and validation of the RRLD model in vitro. b Cell viability of different HaCaT and WS1 cells, as well as primary rat epidermal cells treated with EPI or 5‑Fu after irradiation, as assessed by CCK‑8 assay ( n = 6). c Representative fluorescence microscopy images of reactive oxygen species (ROS) in HaCaT cells treated with EPI or 5‑Fu after irradiation using DCFH‑DA ( n = 3). Scale bar = 100 µm. d Flow cytometry analysis of apoptosis rates in HaCaT cells across different treatment groups and f corresponding quantitative analysis of apoptosis rates ( n = 4). e Flow cytometry analysis of apoptosis rates in WS1 cells across different treatment groups and g corresponding quantitative analysis of apoptosis rates ( n = 4). h Expression levels of apoptosis‑related proteins in different treatment groups (β-actin was the loading control, n = 3) and corresponding quantification of relative Bax/Bcl2 ratio in ( i ) HaCaT and j WS1 cells. Results are presented as mean ± SD. HaCaT cells were treated with 0.05 µM EPI (1 µM 5-Fu), and WS1 and primary rat epidermal cells were treated with 0.05 µM EPI (5 µM 5-Fu) for in vitro RRLD models. For comparisons between multiple groups, one-way ANOVA followed by post-hoc multiple-comparison tests was used. *** P < 0.001

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Biomarker Discovery, In Vitro, Irradiation, Fluorescence, Microscopy, Flow Cytometry, Expressing, Control, Comparison

Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: Identification of key regulatory miRNAs and target genes involved in RRLD by mRNA and miRNA sequencing. a Schematic overview of the study design, including transcriptome (mRNA) and miRNA sequencing followed by integrated bioinformatic analysis. b Venn diagram showing differentially expressed miRNAs among treatment groups ( n = 3). c Venn diagram showing differentially expressed mRNAs among treatment groups ( n = 3). d Relative expression levels of 13 candidate miRNAs in rat skin tissues: comparison between IR and RRLD (EPI or 5‑Fu) groups, as measured by qRT‑PCR (IR: n = 3; RRLD: n = 6). e Relative expression levels of the same candidate miRNAs in human peripheral blood serum samples (RT vs. RRD) measured by qRT‑PCR (IR: n = 17; RRD: n = 8). RT: Radiotherapy; RRD: radiation recall dermatitis. f Relative expression of miR‑338‑3p in HaCaT cells under different treatments ( n = 3). g Relative expression of miR‑338‑3p in WS1 cells under different treatments ( n = 3). h Venn diagram showing that both the human and rat 3′‑UTRs of the candidate genes PTN and TTK harbor predicted binding sites for miR‑338‑3p. i Relative expression levels of PTN and TTK mRNAs in HaCaT cells under different treatments ( n = 3). j Relative expression levels of PTN and TTK mRNAs in WS1 cells under different treatments ( n = 3). k Serum levels of PTN (pg/mL) in HNSCC RT and RRD groups determined by ELISA assay ( n = 3). HNSCC: head and neck squamous cell carcinoma; NPC: Nasopharyngeal carcinoma. Results are presented as mean ± SD. 0.05 µg/µL EPI (10 µg/µL 5-Fu) was used for the in vivo RRLD model. HaCaT cells were treated with 0.05 µM EPI (1 µM 5-Fu), and WS1 was treated with 0.05 µM EPI (5 µM 5-Fu) for in vitro RRLD models. For comparisons between two groups, Student’s t-test was used. For comparisons between multiple groups, one-way ANOVA followed by post-hoc multiple-comparison tests was used. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Sequencing, Expressing, Comparison, Binding Assay, Enzyme-linked Immunosorbent Assay, In Vivo, In Vitro

Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: Regulatory role of miR-338-3p in RRLD models. a Relative miR-338-3p expression levels after transfecting with miR-338-3p mimics ( n = 3). b Relative miR-338-3p expression levels after transfecting with miR-338-3p inhibitor ( n = 3). c Effect of miR-338-3p overexpression on radiosensitivity of HaCaT cells evaluated by colony-formation assay (plating efficiency normalized to negative control, n = 3). d Effect of miR-338-3p knockdown on radiosensitivity of HaCaT cells by colony-formation assay ( n = 3). e Flow cytometry analysis of apoptosis in the RRLD model of HaCaT cells with miR-338-3p mimics and f corresponding quantitative analysis of apoptosis rates ( n = 4). g Flow cytometry analysis of apoptosis in WS1 cells with miR-338-3p mimics and h corresponding quantitative analysis of apoptosis rates ( n = 4). i Study protocol for evaluating effects of miR-338-3p knockdown in RRLD rats. j Representative photographs of rat hindlimb skin showing effects of miR-338-3p knockdown by antagomir in the RRLD model and k corresponding quantitative skin injury scores ( n = 4). l Representative hematoxylin–eosin (H&E) stained images of rat hindlimb skin illustrating effects of miR-338-3p knockdown on histopathology (Epidermal thickness is indicated by area within the white dashed lines, Scar bar = 100 µm) and m corresponding quantitative measurements of epidermal thickness from H&E sections ( n = 4). Results are presented as mean ± SD. 10 µg/µL 5-Fu was used for the in vivo RRLD model. HaCaT cells were treated with 1 µM 5-Fu, and WS1 was treated with 5 µM 5-Fu for in vitro RRLD models. For comparisons between two groups, Student’s t-test was used. ** P < 0.01; *** P < 0.001. NC: negative control

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Over Expression, Colony Assay, Negative Control, Knockdown, Flow Cytometry, Staining, Histopathology, In Vivo, In Vitro

Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: Regulatory role of PTN in RRLD models. a PTN protein expression levels in PTN overexpression or PTN knockdown groups via adenoviral transduction by western blotting (β-actin was the loading control, n = 3) and b corresponding quantification of relative PTN protein expression levels in HaCaT and WS1 cells. c Effect of PTN overexpression (OE) on radiosensitivity of HaCaT cells assessed by colony‑formation assay ( n = 3). d Effect of PTN knockdown on radiosensitivity of HaCaT cells assessed by colony‑formation assay ( n = 3). e Apoptosis rate of HaCaT cells with PTN OE and f corresponding quantitative analysis of apoptosis rates ( n = 4). g Apoptosis rate of WS1 cells with PTN overexpression and h corresponding quantitative analysis of apoptosis rates ( n = 4). i Study protocol for evaluating effects of PTN OE in RRLD rats. j Representative photographs of rat hindlimb skin showing the effects of PTN overexpression (Ad‑PTN OE) in the in vivo RRLD model and k corresponding quantitative skin‑injury scores ( n = 4). l Representative hematoxylin–eosin (H&E) stained images of rat hindlimb skin following PTN overexpression (Epidermal thickness is indicated by area within the white dashed lines, Scar bar = 100 µm) and m corresponding quantitative measurements of epidermal thickness from H&E sections ( n = 4). 10 µg/µL 5-Fu was used for the in vivo RRLD model. HaCaT cells were treated with 1 µM 5-Fu, and WS1 was treated with 5 µM 5-Fu for in vitro RRLD models. Results are presented as mean ± SD. For comparisons between two groups, Student’s t-test was used. ** P < 0.01; *** P < 0.001. NC, negative control; Ad, adenovirus; ns, not significant

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Expressing, Over Expression, Knockdown, Transduction, Western Blot, Control, In Vivo, Staining, In Vitro, Negative Control

Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: miR‑338‑3p participates in the process of RRLD by targeting PTN. a Predicted miR‑338‑3p binding site sequences in the 3′‑UTR of human and rat PTN and Dual‑luciferase reporter assay in WS1 cells co‑transfected with miR‑338‑3p mimics or negative control (NC) and wild‑type (WT) or mutant (MUT) PTN‑3′‑UTR plasmids and b corresponding relative luciferase activity ( n = 4). c The mRNA expression levels of PTN in HaCaT and WS1 cells after transfecting with miR‑338‑3p mimics or ( d ) inhibitors ( n = 3). e The protein expression levels of PTN in HaCaT cells after transfecting with miR‑338‑3p mimics and g corresponding relative PTN protein expression (β-actin as the loading control; n = 3). f The protein expression levels of PTN in HaCaT cells after transfecting with miR‑338‑3p inhibitors and h corresponding relative PTN protein expression (β-actin as the loading control; n = 3). i Representative immunohistochemistry (IHC) images of PTN in non-RRLD rat skin tissue from the miRNA agomir NC group and miR-338-3p agomir group, with ( k ) corresponding immunoreactive scores (blue arrow indicates PTN immunoactivity, n = 4). j Representative IHC images of PTN in non-RRLD rat skin tissue from the miRNA antagomir NC group and miR-338-3p antagomir group, with ( l ) corresponding immunoreactive scores (blue arrow indicates PTN immunoactivity, n = 4). m Flow cytometry analysis of apoptosis in HaCaT and WS1 cells in the RRLD model under different treatments: miRNA mimics NC + Ad‑NC, miR‑338‑3p mimics + Ad‑NC, miRNA mimics NC + Ad‑PTN OE, and miR‑338‑3p mimics + Ad‑PTN OE and corresponding quantitative analysis of apoptosis rates in ( n ) HaCaT and ( o ) WS1 cells ( n = 4). Results are presented as mean ± SD. 10 µg/µL 5-Fu was used for the in vivo RRLD model. HaCaT cells were treated with 1 µM 5-Fu, and WS1 was treated with 5 µM 5-Fu for in vitro RRLD models. For comparisons between two groups, Student’s t-test was used. For comparisons between multiple groups, one-way ANOVA followed by post-hoc multiple-comparison tests was used. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001. Ad, adenovirus; NC, negative control; OE, overexpression

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Binding Assay, Reporter Assay, Negative Control, Mutagenesis, Luciferase, Activity Assay, Expressing, Control, Immunohistochemistry, Flow Cytometry, In Vivo, In Vitro, Comparison, Over Expression

Journal: Molecular Biomedicine

Article Title: miR-338-3p promotes radiation recall like dermatitis by suppressing pleiotrophin via the PI3K/Akt/Bcl2 pathway

doi: 10.1186/s43556-026-00424-5

Figure Lengend Snippet: miR-338-3p targets PTN to induce apoptosis via the PI3K/Akt/Bcl2 signaling pathway. a Representative images of RRLD rat hindlimb skin showing effects of miR‑338‑3p overexpression with or without PTN overexpression in the in vivo RRLD model ( n = 4). b Representative hematoxylin–eosin (H&E) images of RRLD rat hindlimb skin illustrating histopathological changes under the same treatments (Epidermal thickness is indicated by area within the white dashed lines, Scale bar = 100 µm) and c corresponding quantitative skin‑injury scores and d epidermal thickness from H&E images ( n = 4). e Western blot analysis of PTN protein and proteins in the PI3K/Akt/Bcl2 pathway in RRLD (HaCaT) and RRLD (WS1) transfected with miR‑338‑3p inhibitor or mimics, and f corresponding relative protein expression levels of markers in RRLD HaCaT and WS1 cells (β-actin as the loading control; n = 3). g Scheme of miR-338-3p targeting PTN to induce the apoptosis of RRLD in rats. Results are presented as mean ± SD. 10 µg/µL 5-Fu was used for the in vivo RRLD model. HaCaT cells were treated with 1 µM 5-Fu, and WS1 was treated with 5 µM 5-Fu for in vitro RRLD models. For comparisons between two groups, Student’s t-test was used. For comparisons between multiple groups, one-way ANOVA followed by post-hoc multiple-comparison tests was used. * P < 0.05; ** P < 0.01; *** P < 0.001. Ad, adenovirus; NC, negative control; OE, overexpression

Article Snippet: The human skin fibroblast cell line WS1 was obtained from the American Type Culture Collection (ATCC).

Techniques: Over Expression, In Vivo, Western Blot, Transfection, Expressing, Control, In Vitro, Comparison, Negative Control