Journal: International Journal of Molecular Sciences

Article Title: Telmisartan Inhibits Cell Proliferation and Tumor Growth of Esophageal Squamous Cell Carcinoma by Inducing S-Phase Arrest In Vitro and In Vivo

doi: 10.3390/ijms20133197

Figure Lengend Snippet: Effects of telmisartan on proliferation of ESCC cell lines in vitro. Telmisartan inhibited proliferation of ESCC cells. Viability of treated cells differed significantly from that of control cells. ( a ) KYSE150, KYSE180, and KYSE850 cells were seeded in 96-well plate (1.0 × 10 4 cells/well). After 24 h, telmisartan (10, 50, or 100 µM) or DMSO were added to the fresh culture medium. Cell viability was assayed daily from 0 to 48 h. ( b ) Cell viability of ESCC cells at 48 h. (* p < 0.01).

Article Snippet: We used three human ESCC cell lines, KYSE150 (Lot.12112008), KYSE180 (Lot.8012008), KYSE850 (Lot.8252001), which were obtained from Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) on 28 May 2013.

Techniques: In Vitro, Control

Journal: Molecular Cancer

Article Title: Characterization of distinct circular RNA signatures in solid tumors

doi: 10.1186/s12943-022-01546-4

Figure Lengend Snippet: Characterization of circLIFR in solid tumors. A Schematic illustration showing circLIFR is derived from exons 2–5 of LIFR mRNA. The F1/R1 primers were used to measure circLIFR by RT-qPCR. DP, divergent primers; CP, convergent primers. B circLIFR expression measured by RT-qPCR in ESCC, HCC and their matched adjacent normal tissues, normalized to U6 RNA levels. N, normal tissues; T, tumor tissues. Data represent the mean ± S.D., and the p- value was determined by two-tailed paired Student’s t -test. C The presence of circLIFR was validated in KYSE150 cells by RT-PCR. Divergent primers were used to amplify circLIFR from cDNA, but not from genomic DNA. LIFR mRNA linear transcripts were used as a negative control. cDNA, complementary DNA; gDNA, genomic DNA. D Total RNA from KYSE150 cells with or without RNase R treatment was subjected to RT-PCR. E Agarose gel electrophoresis and Sanger sequencing of RT-PCR products of circLIFR in KYSE30 and SK-Hep-1 cells. F RT-qPCR analyses of circLIFR after cytoplasmic/nuclear fractionation of KYSE30 and SK-Hep-1 cells. β-actin mRNA and U6 RNA represent cytoplasmic and nuclear RNAs, respectively. Western blotting confirmed the efficiency of nuclear/cytoplasmic isolation. Data are shown as the mean ± SD. G Fluorescence in situ hybridization of circLIFR in KYSE30 and SK-Hep-1 cells. 18S and U6 RNA represent cytoplasmic and nuclear RNAs, respectively. H circLIFR and LIFR mRNA levels in KYSE30 and SK-Hep-1cells without or with actinomycin D treatment

Article Snippet: BALB/c nude mice were purchased from Beijing HFK Bioscience Co. Ltd. For lung metastasis, 1 × 10 6 KYSE30 cells suspended in 200 μl PBS were injected into the tail vein.

Techniques: Derivative Assay, Quantitative RT-PCR, Expressing, Two Tailed Test, Reverse Transcription Polymerase Chain Reaction, Negative Control, Agarose Gel Electrophoresis, Sequencing, Fractionation, Western Blot, Isolation, Fluorescence, In Situ Hybridization

Journal: Molecular Cancer

Article Title: Characterization of distinct circular RNA signatures in solid tumors

doi: 10.1186/s12943-022-01546-4

Figure Lengend Snippet: circLIFR inhibits cell migration in vitro and tumor metastasis in vivo . A Agarose gel electrophoresis and Sanger sequencing of semi-quantitative RT-PCR products demonstrated that circLIFR was correctly circularized and successfully overexpressed in KYSE30 and SK-Hep-1 cells. B Transwell migration assays showed that circLIFR overexpression inhibited the migratory ability of KYSE30 and SK-Hep-1 cells. C Wound healing assays showed that circLIFR overexpression inhibited cell migration in both KYSE30 and SK-Hep-1 cells. D Decreased tumor metastasis formed in the lungs of mice via tail vein injection of circLIFR-overexpressing KYSE30 cells, as indicated by representative bioluminescent images of lung metastasis at 6 weeks (left) and H&E staining of lung metastatic lesions (right) of mice for each group ( n = 5 mice/group). E Decreased tumor metastasis formed in the livers of mice via spleen injection of circLIFR-overexpressing SK-Hep-1 cells, as indicated by representative bioluminescent images of liver metastasis at 8 weeks (left) and H&E staining of liver metastatic lesions (right) of mice for each group (n = 5 mice/group). F Volcano plot of differentially expressed genes affected by circLIFR in KYSE30 cells. G,H Gene Ontology ( G ) and GSEA analysis ( H ) showed that dysregulated genes upon circLIFR overexpression was involved in cell adhesion. NES, normalized enrichment score; FDR, false discovery rate. p- values are calculated by permutation test. I , J Quantification of metastasis-related genes by RT-qPCR in KYSE30 control cells and circLIFR-overexpressing cells. Data represented mean ± S.D.; the p- values were determined by a two-tailed unpaired Student’s t -test

Article Snippet: BALB/c nude mice were purchased from Beijing HFK Bioscience Co. Ltd. For lung metastasis, 1 × 10 6 KYSE30 cells suspended in 200 μl PBS were injected into the tail vein.

Techniques: Migration, In Vitro, In Vivo, Agarose Gel Electrophoresis, Sequencing, Quantitative RT-PCR, Over Expression, Injection, Staining, Control, Two Tailed Test

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