Journal: Cancers

Article Title: Drug Repurposing Applications to Overcome Male Predominance via Targeting G2/M Checkpoint in Human Esophageal Squamous Cell Carcinoma

doi: 10.3390/cancers14235854

Figure Lengend Snippet: Male ESCC cell lines were more sensitive to decitabine and MK1775 than females. ( a ) Workflow of drug repurposing by using the gene expression data of patients over 60 years old. ( b ) Ten drugs were selected from the drug repurposing results. ( c , d ), Growth curves of KYSE150, KYSE510, KYSE30, and KYSE450 cells were measured by IncuCyte S3 for 72 h. KYSE30 and KYSE450 cells were derived from male patients, while KYSE150 and KYSE510 cells were derived from female patients. Cells were treated with decitabine (10 μM) or MK1775 (200 nM). ( e – g ), Representative image, tumor weights, and tumor volumes of xenografts derived from KYSE30 cells (male), KYSE150 cells (female), and KYSE450 (male) that were treated with decitabine (1.0 mg/kg, i.p.) or MK1775 (60 mg/kg, p.o.). The data shown are the mean ± SD; n = 6 mice per group in KYSE30 and KYSE150; n = 5 mice per group in 450. For tumor weights, data were analyzed using two-tailed t-tests; for tumor volumes, data were analyzed using two-way ANOVA with Bonferroni correction. (* p < 0.05, ** p < 0.01, and *** p < 0.001; ns = not significant).

Article Snippet: A total of 1 × 10 6 KYSE150 cells were subcutaneously injected into 6-week-old female Balb/c nude mice (GemPharmatech Co., Ltd., Nanjing, China).

Techniques: Gene Expression, Derivative Assay, Two Tailed Test

Journal: Cancers

Article Title: Drug Repurposing Applications to Overcome Male Predominance via Targeting G2/M Checkpoint in Human Esophageal Squamous Cell Carcinoma

doi: 10.3390/cancers14235854

Figure Lengend Snippet: Validation RNA-seq proves that MK1775 and decitabine showed a sex-biased treatment response by targeting the G2/M checkpoint. ( a ) Drug-gene network between decitabine and MK1775 and their target genes was assessed in STICH. Gene functional ontology analysis in the ( b ) decitabine KYSE30 cell line, ( c ) MK1775 KYSE30 cell line, and ( d ) MK1775 KYSE150 cell line.

Article Snippet: A total of 1 × 10 6 KYSE150 cells were subcutaneously injected into 6-week-old female Balb/c nude mice (GemPharmatech Co., Ltd., Nanjing, China).

Techniques: Biomarker Discovery, RNA Sequencing, Functional Assay

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: ALDH18A1 knockdown affected cell proliferation, invasion, and migration levels of KYSE150 cells. ALDH18A1 knockdown affected cell proliferation, invasion, and migration levels of KYSE150 cells. ( A ) Bar plot showing the RT-qPCR results of siNC and siALDH18A1 samples. ( B ) Western blot result showing the decreased expression in siALDH18A1 samples. ( C ) Line plot showing the cell proliferation result. ( D & E ) Cellular staining and bar plot showing the cell invasion and migration results; * p -value < 0.05; ** p -value < 0.01; *** p -value < 0.001; Student’s t -test.

Article Snippet: The KYSE150 cell line (Procell Life Science & Technology Co., Ltd., China) was cultured at 37 °C with 5% CO 2 in RPMI1640 with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 U/mL penicillin.

Techniques: Knockdown, Migration, Quantitative RT-PCR, Western Blot, Expressing, Staining

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: ALDH18A1 regulates the expression level of genes in esophageal squamous carcinoma cells (KYSE150 cells). ALDH18A1 regulates the gene expression in KYSE150 cells ( A ) Histogram showed the RT-qPCR results of control and treatment samples. *** p < 0.001. ( B ) ALDH18A1 expression quantified by RNA-seq. Error bars represent mean ± SEM. *** p < 0.001. ( C ) PCA based on FPKM value of all detected genes. The ellipse for each group is the confidence ellipse. ( D ) Volcano plot showing all DEGs between knockdown (KD) and Ctrl samples. P -value < 0.01 and FC (fold change) ≥ 1.5 or ≤ 2/3. ( E ) Hierarchical clustering heat map showing the expression levels of all DEGs. ( F , G ) Top 10 representative GO biological processes of up- or down-regulated genes. ( H ) Bar plot showing the expression pattern and statistical difference of DEGs from RNA sequencing data and RT-qPCR validation. Error bars represent mean ± SEM. *** P -value < 0.001.

Article Snippet: The KYSE150 cell line (Procell Life Science & Technology Co., Ltd., China) was cultured at 37 °C with 5% CO 2 in RPMI1640 with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 U/mL penicillin.

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Control, RNA Sequencing, Knockdown, Biomarker Discovery

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: ALDH18A1 regulates the alternative splicing events of genes in KYSE150 cells. ALDH18A1 regulates the alternative splicing of genes in KYSE150 cells. ( A ) Frequency distribution of different types of ALDH18A1-regulated alternative splicing events. X-axis: RASE number. Y-axis: the different types of AS events. ( B ) Bar plot exhibiting the most enriched GO biological process results of the regulated alternative splicing genes (RASGs). ( C ) Venn diagram showing the overlap genes number of RASGs and DEGs. ( D ) ALDH18A1 regulates alternative splicing of CIRBP. The gene structure and alternative splicing change of the particular ASE responding to ALDH18A1 knock-down (KD) was shown (left). Bottom graph shows the mRNA isoforms/splicing variants structure; the name of each mRNA isoform is labeled on the left. Top-IGV-sashimi plots showing illunima sequencing read density and the number of splicing junction reads. Quantification of the alternative splicing regulation by ALDH18A1 is shown at the right panels. Top-the diagrams depicting the structure of this ASE. The constitutive exon sequences are denoted with white boxes, intron sequences with horizontal line, and alternative exons with blue boxes. RNA-seq quantification (middle) and RT-qPCR validation (bottom) results are shown. The altered ratio of AS events in RNA-seq was calculated using the formula: alternative splice junction reads/(alternative splice junction reads + model splice junction reads). Student’s t -test was performed to compare ALDH18A1-KD and control cells with significance set at a P -value < 0.05. *** P -value < 0.001, ** P -value < 0.01, * P -value < 0.05.

Article Snippet: The KYSE150 cell line (Procell Life Science & Technology Co., Ltd., China) was cultured at 37 °C with 5% CO 2 in RPMI1640 with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 U/mL penicillin.

Techniques: Alternative Splicing, Knockdown, Labeling, Sequencing, RNA Sequencing, Quantitative RT-PCR, Biomarker Discovery, Control

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: ALDH18A1 binds to and regulates the expression of ESCA-associated mRNA in KYSE150 cells. ALDH18A1 binds to mRNAs associated with ESCA in KYSE150 cells. ( A ) WB experiment to verify IP efficiency. ( B ) Pie chart showing the peaks distribution across reference genome. ( C ) Motif analysis showing the top five peaks preferred bound motifs of ALDH18A1 by HOMER software. ( D – E ) Bar plot showing peaks distribution which contains GUAAUCC and UAGCUGG motif. ( F - G ) ALDH18A1 binding peak genes of LAMB3 ( F ), IL1B ( G ). IGV-sashimi plot showing the peaks reads and binding sites across mRNA; the green and red panels represent the position of the peaks (left). Read distribution of bound gene is plotted in the up panel, and the transcripts of each gene are shown below. Quantification of LAMB3 and IL1B expression by RT-qPCR using iRIP-seq data (right). *** P -value < 0.001, ** P -value < 0.01, * P -value < 0.05.

Article Snippet: The KYSE150 cell line (Procell Life Science & Technology Co., Ltd., China) was cultured at 37 °C with 5% CO 2 in RPMI1640 with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 U/mL penicillin.

Techniques: Expressing, Software, Binding Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: ALDH18A1 has carcinogenic functions and regulates alternative splicing events of DNA repair-related genes in esophageal carcinoma cells

doi: 10.1038/s41598-025-08006-1

Figure Lengend Snippet: The si-ALDH18A1 group promotes DNA damage compared with the si-NC group in KYSE150 cells. ( A )The immunofluorescence imaging results of the DNA damage marker protein γ-H2AX, The green signal indicates DNA damage markers.The blue signal is DAPI, staining the nuclei. The Merge image shows the overlay of both channels. ( B )The quantitative comparison results of average fluorescence intensity between the two groups.*** P -value < 0.001.

Article Snippet: The KYSE150 cell line (Procell Life Science & Technology Co., Ltd., China) was cultured at 37 °C with 5% CO 2 in RPMI1640 with 10% fetal bovine serum (FBS), 100 μg/mL streptomycin, and 100 U/mL penicillin.

Techniques: Immunofluorescence, Imaging, Marker, Staining, Comparison, Fluorescence