sw620 human colon cancer cell lines  (ATCC)

Journal: Journal of Advanced Research

Article Title: Activation of sphingosine-1-phosphate receptor 2 (S1PR2) upregulates dihydropyrimidine dehydrogenase (DPD) expression in colon cancer cells

doi: 10.1016/j.jare.2025.01.006

Figure Lengend Snippet: S1PR2 was identified as a mediator of DPD expression in cancer cells. (A) Expression levels of S1PR2 and DPD in human colon cancer tissues (T) and paired adjacent normal tissues (N) (A-i). Statistical analysis showing the correlation between S1PR2 and DPD in cancer tissues (A-ii) but not in adjacent normal tissues (A-iii). P1-P15 are patient codes (n = 15). (B) IHC staining of representative human CRC samples and paired paracancerous tissues. The 5-FU-resistant samples expressed higher levels of S1PR2 and DPD (B-i, left panel) than 5-FU-sensitive samples (B-i, right panel). Both paracancerous tissues paired 5-FU-resistant samples (B-ii, left panel) and 5-FU-sensitive samples (B-ii, right panel) expressed lower levels of S1PR2 and DPD (scale bars: 200 μm and 25 μm). (C) The levels of S1PR2 and DPD in human cancer cell lines (C-i). Statistical analysis showing a correlation of S1PR2 with DPD (C-ii). (D) Comparative analysis of S1PR2 and DPD levels in HT-29 sh-S1PR2 and HT29 sh-control cells (D-i) as well as SW480 S1PR2 and SW480 vector cells (D-ii). The data represent the means ± SD from n = 3 replicates, ** p < 0.01. (E) RT-PCR analysis of the mRNA expression levels of S1PR2 and DPYD in HT-29 sh-S1PR2 and HT29 sh-control cells (E-i) as well as SW480 S1PR2 and SW480 vector cells (E-ii). The data represent the means ± SD from n = 3 replicates. ** p < 0.01. (F) Colony formation and MTT assays for estimating the 5-FU sensitivity of HT-29 sh-S1PR2 and HT-29 sh-control cells (F-i) as well as SW480 S1PR2 and SW480 vector cells (F-ii) (n = 3). (G) FBAL stimulated the expression of S1PR2 and DPD in HT-29 cells (G-i). Silencing of S1PR2 abrogated the stimulation of DPD in HT-29-siS1PR2 cells (G-ii). The data represent the means ± SD from n = 3 replicates, ** p < 0.01.

Article Snippet: Human colon cancer cell lines SW620, HT-29, SW480, RKO, LoVo, HCT116, and HCT-15, the noncancerous cell line CCC-HEL-1, human liver cancer cell line HepG2, as well as human gastric cancer cell lines MGC-803 and BGC-803 were purchased from the China Cell Bank (Shanghai, China) authorized by the American Type Culture Collection (ATCC).

Techniques: Expressing, Immunohistochemistry, Control, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Advanced Research

Article Title: Activation of sphingosine-1-phosphate receptor 2 (S1PR2) upregulates dihydropyrimidine dehydrogenase (DPD) expression in colon cancer cells

doi: 10.1016/j.jare.2025.01.006

Figure Lengend Snippet: TWIST1 acts as a mediator between activated S1PR2 and DPYD. (A-i) The levels of DPD and S1PR2 in SW620 sh-S1PR2 and SW620 sh-control cells. The data represent the means ± SD from n = 3 replicates, ** p < 0.01. (A-ii) Heatmap showing the expression levels of genes that were upregulated in SW620 sh-S1PR2 compared with SW620 sh-control cells according to RNA-seq. (A-iii) Venn diagram showing the number of SW620 sh-S1PR2 genes that were suppressed (red circles) compared with SW620 sh-control (green circles) or upregulated (yellow circles) by SW620 sh-S1PR2 compared with SW620 sh-control (blue circles). (B) GSEA revealed negative enrichment of S1PR2-altered genes in gene sets of drug metabolism enzymes. GSEA plot showing that the S1PR2 level was correlated with the enzymes of 5-FU metabolism (B-i). DPYD was listed among the core enrichment results (B-ii). (C) STRING database analysis showing the functional interactions between S1PR2 and DPD among the 456 differentially expressed genes. Colored lines between proteins indicated various types of interaction evidence. (D) The profiles of TWIST1 expression in various types of tumor samples and paired adjacent tissues in TCGA datasets, including colorectal adenocarcinoma (COAD). Log2 (TPM + 1) was used for log-scale transformation. (E) The correlation between TWIST1 and S1PR2 in COAD tumor analyzed by GEPIA. (F) The levels of S1PR2, TWIST1 and DPD in human colon cancer cell lines (F-i). Scatterplots showing the correlation between TWIST and S1PR2 (F-ii)/DPD (F-iii) in these cancer cell lines. (G) TWIST1 levels in human colon cancer tissues (T) and paired adjacent normal tissues (N) (G-i) (n = 15). Scatterplots showing the correlation between TWIST and S1PR2/DPD in tumor tissues (G-ii, iv) and paired paracancerous tissues (G-iii, v). The data represent the means ± SD; * p < 0.05, ** p < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human colon cancer cell lines SW620, HT-29, SW480, RKO, LoVo, HCT116, and HCT-15, the noncancerous cell line CCC-HEL-1, human liver cancer cell line HepG2, as well as human gastric cancer cell lines MGC-803 and BGC-803 were purchased from the China Cell Bank (Shanghai, China) authorized by the American Type Culture Collection (ATCC).

Techniques: Control, Expressing, RNA Sequencing, Functional Assay, Transformation Assay

Journal: Journal of Advanced Research

Article Title: Activation of sphingosine-1-phosphate receptor 2 (S1PR2) upregulates dihydropyrimidine dehydrogenase (DPD) expression in colon cancer cells

doi: 10.1016/j.jare.2025.01.006

Figure Lengend Snippet: Activation of S1PR2 promoted the binding of TWIST1 to the DPYD promoter. (A and B) TWIST1 protein and mRNA levels in HT-29 sh-S1PR2 and HT29 sh-control cells (A) as well as SW480 S1PR2 and SW480 vector cells (B). The data represent the means ± SD from n = 3 replicates. ** p < 0.01. (C) The levels of TWIST1 in the nuclei and cytoplasm of HT-29 (left) and SW480 S1PR2 cells (right) with and without JTE013 treatment. H3 and GAPDH were employed as specific nuclear and cytoplasmic protein loading controls, respectively. (D) The protein (D-i) and mRNA (D-ii) levels of DPD in HT-29 cells with silenced TWIST1 expression. The data represent the means ± SD form n = 3 replicates, ** p < 0.01. (E) HPLC-UV analysis intracellular 5-FU levels in HT-29 cells (blue line in E-i), 5-FU-treated HT-29-siNC cells (green line in E-i) and 5-FU-treated HT-29-siTWIST1 cells (red line in E-i). Statistical analysis indicated higher 5-FU levels in HT-29-siTWIST1 cells compared to HT-29-siNC cells (E-ii). The data represent the means ± SD from n = 3 replicates, ** p < 0.01. (F) TWIST1 response element motif sequences (F-i). Bioinformatic analysis of the potential TWIST1-binding sites in the 2 kb DPYD promoter region using the Jaspar database (F-ii). Luciferase assay of DPYD promoter activity. Co-transfection of HT-29 cells with TWIST1 and DPYD promoter pGL3-Luciferase constructs (F-iii). (G) ChIP assay showing the direct interaction of TWIST1 with the DPYD promoter in HT-29 cells. IgG served as a negative control (n = 3). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Human colon cancer cell lines SW620, HT-29, SW480, RKO, LoVo, HCT116, and HCT-15, the noncancerous cell line CCC-HEL-1, human liver cancer cell line HepG2, as well as human gastric cancer cell lines MGC-803 and BGC-803 were purchased from the China Cell Bank (Shanghai, China) authorized by the American Type Culture Collection (ATCC).

Techniques: Activation Assay, Binding Assay, Control, Plasmid Preparation, Expressing, Luciferase, Activity Assay, Cotransfection, Construct, Negative Control

Journal: Journal of Advanced Research

Article Title: Activation of sphingosine-1-phosphate receptor 2 (S1PR2) upregulates dihydropyrimidine dehydrogenase (DPD) expression in colon cancer cells

doi: 10.1016/j.jare.2025.01.006

Figure Lengend Snippet: The S1PR2-activated TWIST1 bound to the DPYD promoter via the Hippo/TEAD1-TWIST1 pathway. (A-C) The levels of p-LATS/LATS and p-YAP1/YAP1 in HT-29 sh-S1PR2 and HT29 sh-control cells (A), SW620 sh-S1PR2 and SW620 sh-control cells (B), as well as SW480 S1PR2 and SW480 vector cells (C). (D) Subcellular distribution analysis of p-YAP1 and YAP1 in HT-29 cells with or without JTE013 treatment. H3 and GAPDH were employed as specific nuclear and cytoplasmic protein loading controls, respectively. (E and F) The levels of LATS, YAP1, TWIST1 and DPD in HT-29-siLATS cells (E) and HT-29-siYAP1 cells (F). The data represent the means ± SD from n = 3 replicates, ** p < 0.01. (G) The mRNA levels of YAP1 (G-left) and TWIST1 (G-right) in HT-29 siYAP1 cells and HT-29 siNC cells. The data represent the means ± SD. n = 3. ** p < 0.01. (H) ChIP assay using a specific primer for TWIST1-3 (H-i) showed the association between YAP1/TEAD1 and TWIST1 in HT-29 cells (H-ii). IgG served as a negative control (n = 3). (I) The levels of p-LATS/LATS and p-YAP1/YAP1 in HT-29 (I-i), HT-29-siNC and HT-29-siS1PR2 cells (I-ii) exposed to FBAL. Data are presented as means ± SD from n = 3 replicates, ** p < 0.01.

Article Snippet: Human colon cancer cell lines SW620, HT-29, SW480, RKO, LoVo, HCT116, and HCT-15, the noncancerous cell line CCC-HEL-1, human liver cancer cell line HepG2, as well as human gastric cancer cell lines MGC-803 and BGC-803 were purchased from the China Cell Bank (Shanghai, China) authorized by the American Type Culture Collection (ATCC).

Techniques: Control, Plasmid Preparation, Negative Control