colorectal carcinoma hct116 cell line  (ATCC)

Journal: bioRxiv

Article Title: Long-chain Polyunsaturated Fatty Acids Mitigate In Vitro Skeletal Muscle Wasting Induced by Colorectal Carcinoma Cells via a 15-LOX-dependent Pathway

doi: 10.64898/2026.05.22.726995

Figure Lengend Snippet: (A) : Day 3 post-differentiation C2C12 myotubes were treated with fresh DM, CT26 CM, DM containing 50 µM NDGA, or 10 µM BLX-3887 for 72 hours. C2C12 myotubes were fixed and stained for sarcomeric myosin (MF20c, green) and myogenin (F5Dc, red). Nuclei was counter stained by DAPI (blue). (B) : Myotube diameter for C2C12 myotubes was quantified as described in Methods. Groups labeled with different letters are significantly different from one another, while groups sharing a common letter are not significantly different.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and colorectal carcinoma cell line CT26 (ATCC, CRL-2638) were cultured separately in growth media (GM) consisting of Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in cell incubator with 5% CO 2 .

Techniques: Staining, Labeling

Journal: bioRxiv

Article Title: Long-chain Polyunsaturated Fatty Acids Mitigate In Vitro Skeletal Muscle Wasting Induced by Colorectal Carcinoma Cells via a 15-LOX-dependent Pathway

doi: 10.64898/2026.05.22.726995

Figure Lengend Snippet: Day 3 post-differentiation C2C12 myotubes were treated with CT26 conditioned media (CM) with or without the pan-LOX inhibitor nordihydroguaiaretic acid (NDGA) (50 µM) or the 15-LOX specific inhibitor BLX3887 (10 µM). Then, individual LC-PUFAs including ARA, EPA, DHA, and DPA were spiked into the wells to reach a final concentration of 25 µM. After 72 hours, C2C12 myotubes were fixed in 4% paraformaldehyde (PFA) in preparation for immunocytochemistry analysis. (A) C2C12 myotubes were stained for sarcomeric myosin (MF20c, green) and myogenin (F5Dc, red). Nuclei was counter stained by DAPI (blue). (B-C) Myotube diameter for C2C12 myotubes with or without NDGA (B) or BLX-3887 (C) was quantified as described in Methods. Scale bar is 200 μm. *P < 0.05 for difference of PUFA treatments vs. vehicle, and #P < 0.05 for difference between respective LC-PUFA treatments with or without LOX inhibitors. (D-E) C2C12 myoblasts were differentiated for 3 days and then exposed to CT26 CM and individual PUFAs including ARA, EPA, DPA, and DHA for 3 hours. RNA was extracted from mature myotubes. mRNA expression fold change of Il6 (D) and Ccl2 (E) in response to CT26 CM and PUFA treatments was shown. *P < 0.05 for effects of PUFA treatments, and #P < 0.05 for difference between DM and CT26 CM.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and colorectal carcinoma cell line CT26 (ATCC, CRL-2638) were cultured separately in growth media (GM) consisting of Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in cell incubator with 5% CO 2 .

Techniques: Concentration Assay, Immunocytochemistry, Staining, Expressing

Journal: bioRxiv

Article Title: Long-chain Polyunsaturated Fatty Acids Mitigate In Vitro Skeletal Muscle Wasting Induced by Colorectal Carcinoma Cells via a 15-LOX-dependent Pathway

doi: 10.64898/2026.05.22.726995

Figure Lengend Snippet: Co-cultures of mature C2C12 myotubes and CT26 carcinoma cells were treated with individual LC-PUFAs including ARA, EPA, DPA, and DHA (25 µM). After 72 hours, conditioned media from the lower compartments was collected and C2C12 myotubes were fixed and stained for sarcomeric myosin (MF20c, green) and myogenin (F5Dc, red). Nuclei was counter stained by DAPI (blue) (A) . Myotube diameter for C2C12 myotubes in the base plates was quantified as described in Methods (B) . Scale bar is 200 μm. Groups labeled with different letters are significantly different from one another, while groups sharing a common letter are not significantly different. (C- G) Conditioned media from C2C12-CT26 co-culture was analyzed by targeted Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to profile lipid metabolites in response to PUFA treatments. (C) : A heatmap of the top 50 most differentially regulated lipid mediators detected in conditioned culture media samples from C2C12 myotubes. (D-G) : Concentration of representative lipid mediator metabolites downstream of arachidonic acid (ARA) (e.g., PGE 2 , 15-HETE, and LXA 4 ) (D) , EPA (e.g., 15-HEPE, RvE1) (E), DPA (e.g., RvD5 n-3 DPA ) (F) , and DHA (e.g., 14-HDoHE, MaR2, 17-HDoHE, and RvD1) (G) . Bars show the mean ± SEM of media from 3 wells (biological replicates). P-values were determined by two-tailed unpaired t-tests. ∗p < 0.05, **p<0.01, ***p<0.001, and ****p<0.0001 vs. C2C12 myotubes without PUFAs or CT26 inserts.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and colorectal carcinoma cell line CT26 (ATCC, CRL-2638) were cultured separately in growth media (GM) consisting of Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in cell incubator with 5% CO 2 .

Techniques: Staining, Labeling, Co-Culture Assay, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Long-chain Polyunsaturated Fatty Acids Mitigate In Vitro Skeletal Muscle Wasting Induced by Colorectal Carcinoma Cells via a 15-LOX-dependent Pathway

doi: 10.64898/2026.05.22.726995

Figure Lengend Snippet: Concentration of lipid mediator metabolites downstream of arachidonic acid (ARA) (e.g., 5-HETE, 6-keto-PGF 1α , PGD 2 , and PGF 2α ) (A) , EPA (e.g., 18-HEPE, RvE2) (B), DPA (e.g., MaR1 n-3 DPA , PD1 n- 3 DPA ) (C) , and DHA (e.g., 4-HDoHE, 7-HDoHE, PD1, PDX, RvD2, RvD3, RvD5, RvD6, AT-RvD3, AT-RvD6) (D) Bars show the mean ± SEM of media from 3 wells (biological replicates). P-values were determined by two- tailed unpaired t-tests. ∗p < 0.05, **p<0.01, ***p<0.001, and ****p<0.0001 vs. C2C12 myotubes without PUFAs or CT26 inserts.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and colorectal carcinoma cell line CT26 (ATCC, CRL-2638) were cultured separately in growth media (GM) consisting of Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in cell incubator with 5% CO 2 .

Techniques: Concentration Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Long-chain Polyunsaturated Fatty Acids Mitigate In Vitro Skeletal Muscle Wasting Induced by Colorectal Carcinoma Cells via a 15-LOX-dependent Pathway

doi: 10.64898/2026.05.22.726995

Figure Lengend Snippet: (A) : Day 3 post-differentiation C2C12 myotubes were treated with 100 ng/mL TNFα (top) or CT26 CM (bottom) in the presence or absence of 100 nM of individual mature SPMs including RvD1, RvD5, MaR1, PD1, RvE1, LXA 4 , and RvD2 n-3 DPA for 72 hours. Resulting C2C12 myotubes were fixed and stained for sarcomeric myosin (MF20c, green) and myogenin (F5Dc, red). Nuclei was counter stained by DAPI (blue). (B- C) Myotube diameter for C2C12 myotubes receiving TNFα (B) or CT26 CM (C) was quantified as described in Methods. Scale bar is 200 μm. *P < 0.05 for difference compared to C2C12 myotubes receiving TNFα or CT26 CM alone. (D-E) C2C12 myoblasts were differentiated for 3 days and exposed to TNFα (100 ng/mL) with or without individual SPMs including RvD1, RvD5, MaR1, PD1, RvE1, LXA 4 , and RvD2 n-3 DPA (100nM) for 3 hours, after which RNA was extracted from mature myotubes. mRNA expression was determined by RT-qPCR and fold change of Il6 (D) and Fbxo32 (E) in response to TNFα exposure and SPM treatments was shown. *P < 0.05 for difference compared to C2C12 myotubes receiving TNFα.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and colorectal carcinoma cell line CT26 (ATCC, CRL-2638) were cultured separately in growth media (GM) consisting of Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in cell incubator with 5% CO 2 .

Techniques: Staining, Expressing, Quantitative RT-PCR

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: 10-HDA inhibited CRC cell growth and gap closure and increased intracellular ROS. ( A ) MTT analysis of HCT 116 and HT-29 cell viability after 48 h exposure to 10-HDA. ( B ) Colony formation of HCT 116 cells following 14 d treatment with 10-HDA. ( C ) Scratch assay analysis of HCT 116 gap closure after 24 h incubation with 10-HDA. ( D ) Flow cytometric analysis of HCT 116 intracellular ROS levels following 3 h exposure to 10-HDA. ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no statistical significance. Control, vehicle-treated cells.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Wound Healing Assay, Incubation, Control

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: 10-HDA inhibited tumor growth and modified the histopathological features of HCT 116 xenografts in vivo. ( A ) Schematic overview of HCT 116 xenograft establishment and 10-HDA administration. ( B ) Representative images of xenograft tumors from saline-treated control mice and 10-HDA-treated mice, with corresponding tumor volume quantification. Control, blue; 100 mg/kg 10-HDA, orange; 200 mg/kg 10-HDA, purple. ( C ) H&E-stained sections of xenograft tumors. Red arrows indicate tumor cells; yellow arrows denote necrotic regions; blue arrows indicate atypical (pathological) mitotic figures; and green arrows indicate inflammatory cell infiltration. ( D ) Ki-67 immunohistochemical staining of xenograft tumors from control and 10-HDA-treated mice. n = 6; * p < 0.05, ** p < 0.01, **** p < 0.0001; ns, no statistical significance. Control, saline-treated tumor-bearing mice.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Modification, In Vivo, Saline, Control, Staining, Immunohistochemical staining

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: Evaluation of the safety profile of 10-HDA treatment. ( A ) Mice weight changes compared with normal mice. Normal, red; model, blue; 100 mg/kg 10-HDA, orange; 200 mg/kg 10-HDA, purple. ( B ) Serum levels of liver function markers (AST, ALT, and ALP). ( C ) Representative H&E-stained histopathological sections of the heart, liver, spleen, lung and kidney. All analyses were performed on samples from healthy mice (normal control), and saline (model)- and 10-HDA-treated mice bearing HCT 116 xenograft tumors. n = 6; * p < 0.05, ** p < 0.01; ns, no statistical significance.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Staining, Control, Saline

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: Transcriptomic analysis of ectopic xenograft tumor tissues derived from HCT 116 cells in 10-HDA- and saline-treated (control) mice. ( A ) Volcano plot of DEGs. Each dot represents one gene; red indicates significantly upregulated genes and green indicates significantly downregulated genes (FDR < 0.05 and |log2FoldChange| > 1). ( B ) Heatmap of representative DEGs related to tumor progression. Colors indicate relative expression (red, upregulation; blue, downregulation). ( C ) GO enrichment of DEGs showing top terms in biological process (BP, red), cellular component (CC, green), and molecular function (MF, blue). ( D ) Top 20 enriched KEGG pathways of DEGs. The x-axis shows the number of DEGs in each pathway; bar color represents statistical significance (adjusted p -value). ( E ) Relative mRNA expression levels of ANO1 , MKi67 , LTBP1 , FGF19 , NECTIN4 , PLD1 , and SKA1 in HCT 116 cells treated with 10-HDA or vehicle (control). Control, dark blue; 1.0 mM 10-HDA, blue; 1.5 mM 10-HDA, light blue; 2.0 mM 10-HDA, green. ( F ) Relative mRNA expression levels of ANO1 , MKi67 , LTBP1 , FGF19 , NECTIN4 , PLD1 , and SKA1 in xenograft tumors from 10-HDA- or saline-treated (control) mice. Control, green; 100 mg/kg 10-HDA, blue; 200 mg/kg 10-HDA, yellow. * p < 0.05, ** p < 0.01, **** p < 0.0001; ns, no statistical significance.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Derivative Assay, Saline, Control, Expressing

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: 10-HDA induced apoptosis in HCT 116 cells and subcutaneous HCT 116 xenograft tumors. ( A ) Relative mRNA expression levels of Bcl-2 , BAX , BINP2 , and caspase-3 in HCT 116 cells treated with 10-HDA or vehicle (control). Control, dark blue, 1.0 mM 10-HDA, blue; 1.5 mM 10-HDA, light blue; 2.0 mM 10-HDA, green. ( B ) Relative mRNA expression levels of Bcl-2 , BAX , BINP2 , and caspase-3 in xenograft tumors from 10-HDA- or saline-treated (control) mice. Control, green; 100 mg/kg 10-HDA, blue; 200 mg/kg 10-HDA, yellow. ( C ) Western blotting analysis of caspase-3, Bcl-2, BAX, and cleaved caspase-3 protein levels, as well as the calculated cleaved caspase-3/caspase-3 and Bcl-2/BAX ratios in HCT 116 cells treated with 10-HDA or vehicle (control). ( D ) Immunohistochemical staining of Bcl-2 and BAX in xenograft tumors from 10-HDA- or saline-treated (control) mice. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no statistical significance.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Expressing, Control, Saline, Western Blot, Immunohistochemical staining, Staining

Journal: Foods

Article Title: 10-Hydroxy-2-decenoic Acid Suppresses Colorectal Cancer Progression by Inhibiting Wnt/β-Catenin Signaling and Promoting Apoptosis

doi: 10.3390/foods15091608

Figure Lengend Snippet: Involvement of Wnt/β-Catenin signaling in the anti-CRC effects of 10-HDA. ( A ) Relative mRNA expression levels of WNT11 , RNF43 , FRAT1 , β-catenin , and MMP7 in HCT 116 cells treated with 10-HDA or vehicle (control). Control, dark blue; 1.0 mM 10-HDA, blue, 1.5 mM 10-HDA, light blue; 2.0 mM 10-HDA, green. ( B ) Relative mRNA expression levels of WNT11 , RNF43 , FRAT1 , β-catenin , and MMP7 in xenograft tumors from 10-HDA- or saline-treated (control) mice. Control, green; 100 mg/kg 10-HDA, blue; 200 mg/kg 10-HDA, yellow. ( C ) Western blotting analysis of β-catenin expression in HCT 116 cells treated with 10-HDA or vehicle (control). ( D ) Western blotting analysis of β-catenin, p-GSK3β and GSK3β in xenograft tumor tissues from 10-HDA- and saline-treated (control) mice. ( E ) Immunohistochemical staining of β-catenin in xenograft tumors. ( F ) Western blotting analysis and densitometric quantification of the nuclear and cytoplasmic β-catenin in HCT 116 cells after 10-HDA or vehicle (control) treatment. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; ns, no statistical significance.

Article Snippet: Human colorectal carcinoma cell lines HCT 116 (ATCC; CCL-247) and HT-29 (ATCC; HTB-38) were purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Expressing, Control, Saline, Western Blot, Immunohistochemical staining, Staining