Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Validation of selected miR-218-5p Gene Targets. ( A ) Validation of BIRC5 and DDX21 as bona fide gene targets for miR-218-5p in CRC. Two-tailed t -test was used to compare different groups. ** p < 0.005, *** p < 0.0005, **** p < 0.00005. ( B ) Western blot showing DDX21 protein expression in miR-218-5p overexpressing compared to control HT-29 and HCT116 cells. Quantification of DDX21 protein expression normalized to ACTB is shown in the right panel. ( C ) Gene effect score based on CRISPR-Cas9 screen data in 40 CRC cell models from the DepMap database.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Biomarker Discovery, Two Tailed Test, Western Blot, Expressing, Control, CRISPR

Journal: International Journal of Molecular Sciences

Article Title: Epigenetic Silencing of miR-218-5p Modulates BIRC5 and DDX21 Expression to Promote Colorectal Cancer Progression

doi: 10.3390/ijms26094146

Figure Lengend Snippet: Suppression of SLIT2 and SLIT3 miR-218 Host Genes in CRC. ( A ) Genomic location of miR-218-1 and miR-218-2 within the SLIT2 and SLIT3 genomic region on chromosome 4 and 5, respectively. ( B ) Correlation plot between miR-218-5p and SLIT2 ( left ) and SLIT3 ( right ) in a large cohort of COAD (n = 450) from the ENCORI project. ( C ) Downregulation of SLIT2 ( left ) and SLIT3 ( right ) in COAD (n = 275) compared to normal colon tissue (n = 349) from GEPIA2 database. T: tumor, N: normal. * p < 0.05. ( D ) Methylation analysis of SLIT2 and SLIT3 promoters using bisulfite conversion and NGS in a panel of CRC cell models (HCT116, HT-29, SW-480, LoVo, and DLD-1) compared to the MCF10A normal epithelial cells. ( E ) Schematic representation illustrating (🠯) downregulation of SLIT2 and SLIT3 in CRC to lead to miR-218-5p suppression (🠯), thus promoting tumorigenesis due to lifted suppression (🠭) of BIRC5, DDX21, and other gene targets identified in the current study.

Article Snippet: A notable observation within SLIT2 , which encodes miR-218-1 within its intronic region, is the detection of higher levels of hypermethylation near the end of the promoter regions in the CRC cell lines, just upstream of the transcription start site, where the TATA box is typically located.

Techniques: Methylation

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Antitumor effect in LIM1215(A) xenografts by treatment received. (A and B) assessed by average tumor volume and (C) relative growth rate. Arrowheads indicate dosing. Arrows indicate sample harvesting. Data represent mean ± SE. ( n = 3–7. * P < .05). B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Outcome of immunohistochemistry for Ki-67 in LIM1215(B) xenograft sections. (A) Using vehicle control, (B) panitumumab-bevacizumab, (C) bevacizumab-panitumumab, and (D) bevacizumab-bevacizumab. (E) Proportion of Ki-67-positive cells in all treatment groups. Sections were IHC stained for Ki-67 (brown) and counterstained with hematoxylin (purple). Representative images of the sections are shown. Data in the graph represent the mean ± SE ( n = 6–8). ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Immunohistochemistry, Control, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Levels of Phosphorylated Growth Factor Receptors in LIM1215(A) Xenografts Treated with PB, BP, and BB Relative to Vehicle Control

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Results of western blotting in LIM1215(B) xenografts. (A) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-panitumumab. (B) EPHA2 and pEPHA2 with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (C) Phosphorylation of EPHA2 for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (D) compared with bevacizumab-bevacizumab. (E) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-panitumumab and (F) RSK and pRSK with panitumumab-bevacizumab compared with bevacizumab-bevacizumab. (G) Phosphorylation of RSK for panitumumab-bevacizumab compared with bevacizumab-panitumumab and (H) compared with bevacizumab-bevacizumab. Data represent mean ± SD ( n = 8). ** P < .01, *** P < .001. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Western Blot, Phospho-proteomics, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Enrichment Analysis of LIM1215(A) Xenografts Treated with Bevacizumab-Bevacizumab Compared with Vehicle Control (all Canonical Pathways, P < .001)

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Control, Activation Assay, Inhibition

Journal: Neoplasia (New York, N.Y.)

Article Title: Biologic Response of Colorectal Cancer Xenograft Tumors to Sequential Treatment with Panitumumab and Bevacizumab

doi: 10.1016/j.neo.2018.04.006

Figure Lengend Snippet: Relative expression of (A–H) lipogenic ( FASN, HMGCR, MVD, LSS ) and (I–L) hypoxia-related ( CA9, TGFBI ) genes in LIM1215(B) xenograft tumors. Expression relative to vehicle control with first-line treatment is shown in (A–D) and (I–J), and with sequential treatment in (E–H) and (K–L). Data represent mean ± SD ( n = 8). * P < .05, ** P < .01. B, bevacizumab; P, panitumumab; V, vehicle control.

Article Snippet: The human colon cancer cell line LIM1215 was obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK).

Techniques: Expressing, Control

Journal: Journal of Clinical Laboratory Analysis

Article Title: Hsa_hsa_circ_0081069 promotes the progression of colorectal cancer through sponging miR ‐665 and regulating E2F3 expression

doi: 10.1002/jcla.24710

Figure Lengend Snippet: Hsa_circ_0081069 was upregulated in CRC tissues and cells. (A) Schematic illustration of Hsa_circ_0081069 structure. (B) qRT‐PCR analysis of hsa_circ_0081069 expression in CRC cell lines and FHC cells. (C) Cells were treated with Actinomycin D, and the level of Hsa_circ_0081069 and COL1A2 mRNA at different time points were determined by qRT‐PCR. (D) Expression level of hsa_circ_0081069 and COL1A2 mRNA after RNase R treatment was determined by qRT‐PCR. (E) Hsa_circ_0081069 expression was determined by qRT‐PCR in CRC tissues and para‐cancerous tissues. (F) KM‐plotter analysis of the overall survival of CRC patients in Hsa_circ_0081069 high and low expression group. *** p < 0.001

Article Snippet: CRC cell lines were acquired from Shanghai Fuxiang Biotechnology Co., Ltd, and cultivated in RPMI‐1640 medium supplemented with 10% fetal bovine serum (FBS, Hyclone) and 1% penicillin/streptomycin (Hyclone) in a humidified incubator at 37°C with 5% CO 2 . miR‐665 mimic/inhibitor and negative controls (miR‐NC or NC inhibitor), and pcDNA.3.1 vector and pcDNA3.1‐E2F3 expression vector were purchased from RiboBio Co. Ltd.

Techniques: Quantitative RT-PCR, Expressing

Journal: Nutrients

Article Title: Docosahexaenoic Acid Coordinating with Sodium Selenite Promotes Paraptosis in Colorectal Cancer Cells by Disrupting the Redox Homeostasis and Activating the MAPK Pathway

doi: 10.3390/nu16111737

Figure Lengend Snippet: The SDCT could induce cell death in CRC cells. ( A , B ) SW620 and RKO cells were exposed to sodium selenite for 24 h and then the cell viability was determined by the CCK8 assay, as described in the . ( C , D ) SW620 and RKO cells were exposed to DHA for 24 h, and then the cell viability was determined by the CCK8 assay. ( E – H ) SW620 and RKO cells were exposed to sodium selenite and DHA for 24 h, and then the cell viability was determined by the CCK8 assay. The dose–response matrix and synergy score were determined by SynergyFinder 2.0. ( I ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h and subjected to Calcein-AM/PI staining (Calcein AM: live cells; PI: dead cells). Scale bar: 400 µm. The representative images are shown. The corresponding quantitative histograms from three independent experiments were shown (The values represent the mean ± SD, n = 3. Two-tailed unpaired Student’s t -test. ns: not significant; ** p < 0.01).

Article Snippet: The human CRC cell line SW620 (Rosetta Stone Biotechnology, Jinan, China) was cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA).

Techniques: CCK-8 Assay, Staining, Two Tailed Test

Journal: Nutrients

Article Title: Docosahexaenoic Acid Coordinating with Sodium Selenite Promotes Paraptosis in Colorectal Cancer Cells by Disrupting the Redox Homeostasis and Activating the MAPK Pathway

doi: 10.3390/nu16111737

Figure Lengend Snippet: DHA remodels the redox state within cancer cells. ( A – C ) SW620 cells were exposed to DHA for 24 h, and the GSH/GSSG ratio was determined by the methods described in the . ( D ) SW620 cells were exposed to DHA for 24 h, and MDA was determined by the methods described in the . ( E , F ) Assessment of indicated protein levels using Western blotting in SW620 cells exposed to DHA. The corresponding quantitative histograms from three independent experiments were shown (The values represented the mean ± SD, n = 3. Two-tailed unpaired Student’s t -test or one-way ANOVA. ns: not significant; * p < 0.05; ** p < 0.01; **** p < 0.0001).

Article Snippet: The human CRC cell line SW620 (Rosetta Stone Biotechnology, Jinan, China) was cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA).

Techniques: Western Blot, Two Tailed Test

Journal: Nutrients

Article Title: Docosahexaenoic Acid Coordinating with Sodium Selenite Promotes Paraptosis in Colorectal Cancer Cells by Disrupting the Redox Homeostasis and Activating the MAPK Pathway

doi: 10.3390/nu16111737

Figure Lengend Snippet: The SDCT disrupts redox homeostasis. ( A , B ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h, and the GSH/GSSG ratio was determined. ( C ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h, and MDA was determined. ( D ) Assessment of indicated protein levels using Western blotting in SW620 cells exposed to 1.5 µM sodium selenite and 25 µM DHA for 24 h. ( E , F ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 12 h, and the intracellular ROS level was measured by flow cytometry and a fluorescent microscope. Scale bar: 100 µm. ( G ) The viability of SW620 cells exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h with a pretreatment of 5 mM NAC (3 h). The representative images and the corresponding quantitative histograms from three independent experiments were shown (The values represent the mean ± SD, n = 3. Two- tailed unpaired Student’s t -test. * p < 0.05; *** p < 0.001; **** p < 0.0001 versus control).

Article Snippet: The human CRC cell line SW620 (Rosetta Stone Biotechnology, Jinan, China) was cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA).

Techniques: Western Blot, Flow Cytometry, Microscopy, Two Tailed Test, Control

Journal: Nutrients

Article Title: Docosahexaenoic Acid Coordinating with Sodium Selenite Promotes Paraptosis in Colorectal Cancer Cells by Disrupting the Redox Homeostasis and Activating the MAPK Pathway

doi: 10.3390/nu16111737

Figure Lengend Snippet: The SDCT activates MAPKs and induces paraptosis. ( A ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h and observed by light microscopy. The arrows in red indicate cytoplasmic vacuoles. Scale bar: 100 µm. ( B ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA for 12 h and observed by transmission electron microscopy. The L in red indicates lysosomes. The V in red indicates cytoplasmic vacuoles. The N in red indicates a cell nucleus. Scale bar: 2 µm. ( C ) The viability of SW620 cells exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h with a pretreatment of CHX (2 h). ( D ) Assessment of indicated protein levels using Western blotting in SW620 cells exposed to 11.5 µM sodium selenite and 25 µM DHA for 24 h. The representative images and the corresponding quantitative histograms from three independent experiments were shown. (The values represented the mean ± SD, n = 3. Two-tailed unpaired Student’s t -test. *** p < 0.001 versus control).

Article Snippet: The human CRC cell line SW620 (Rosetta Stone Biotechnology, Jinan, China) was cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA).

Techniques: Light Microscopy, Transmission Assay, Electron Microscopy, Western Blot, Two Tailed Test, Control

Journal: Nutrients

Article Title: Docosahexaenoic Acid Coordinating with Sodium Selenite Promotes Paraptosis in Colorectal Cancer Cells by Disrupting the Redox Homeostasis and Activating the MAPK Pathway

doi: 10.3390/nu16111737

Figure Lengend Snippet: The redox state dynamically activates the MAPK system. ( A ) Assessment of indicated protein levels and quantitative analysis using Western blotting in SW620 cells exposed to 11.5 µM sodium selenite and 25 µM DHA. ( B – D ) SW620 cells were exposed to 11.5 µM sodium selenite and 25 µM DHA, and the GSH/GSSG ratio was determined by the methods described in the . ( E ) Assessment of indicated protein levels and quantitative analysis using Western blotting in SW620 cells exposed to 11.5 µM sodium selenite and 25 µM DHA. ( F ) Model of cell paraptosis induced by a combination of sodium selenite and DHA. The combined intervention produces excess ROS, inhibits Nrf2/HO-1, destroys redox homeostasis, and finally activates ERK1/2, inducing the paraptosis of CRC cells. The corresponding quantitative histograms from three independent experiments were shown (The values represented the mean ± SD, n=3. One-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 versus the control).

Article Snippet: The human CRC cell line SW620 (Rosetta Stone Biotechnology, Jinan, China) was cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA).

Techniques: Western Blot, Control