Journal: American Journal of Cancer Research

Article Title: Activation of the IL-17 signalling pathway by the CXCL17-GPR35 axis affects drug resistance and colorectal cancer tumorigenesis

doi:

Figure Lengend Snippet: Chemokine CXCL17 and its receptor GPR35 exhibit higher levels in CRC drug-resistant tissues and cells. A. Differential gene volcano plot between OXA-sensitive and OXA-resistant groups in CRC tissues; B. Box plot of CXCL17 expression in OXA-sensitive and OXA-resistant tumor tissues; C. Western-blot detection of CXCL17 and GPR35 protein expression levels in FHC and HCT15 cells; D. RT-qPCR detection of mRNA expression of CXCL17 and GPR35. **P < 0.01, compared with the FHC group.

Article Snippet: Drug-resistant human colorectal cancer cells (HCT15) were cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum, 1% penicillin and streptomycin, and 500 ng/ml taxol (a microtubule-stabilising drug used to treat cancer) (Med Chem Express, Monmouth Junction, NJ, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: American Journal of Cancer Research

Article Title: Activation of the IL-17 signalling pathway by the CXCL17-GPR35 axis affects drug resistance and colorectal cancer tumorigenesis

doi:

Figure Lengend Snippet: CXCL17 plays a critical role in proliferation and drug-resistance of HCT15 cells. A, B. CCK-8 assay was performed to detect cell viability after the treatment with CXCL17 silence or different concentrations of taxol; C. Wound-healing assay for cell migration ability of HCT15 cells upon siNC or si-CXCL17 transfection; D. Transwell assay was applied to analyze cell invasion ability; E. Cell cycle was detected by flow cytometry; F. Cell apoptosis was detected by flow cytometry. Scale bar was 50 μm. Compared with siNC group, *P < 0.05; **P < 0.01.

Article Snippet: Drug-resistant human colorectal cancer cells (HCT15) were cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum, 1% penicillin and streptomycin, and 500 ng/ml taxol (a microtubule-stabilising drug used to treat cancer) (Med Chem Express, Monmouth Junction, NJ, USA).

Techniques: CCK-8 Assay, Wound Healing Assay, Migration, Transfection, Transwell Assay, Flow Cytometry

Journal: American Journal of Cancer Research

Article Title: Activation of the IL-17 signalling pathway by the CXCL17-GPR35 axis affects drug resistance and colorectal cancer tumorigenesis

doi:

Figure Lengend Snippet: IL-17 signaling pathway is involved in tumorigenesis and drug resistance of HCT15 cells. A. CCK-8 assay was performed to detect cell viability of HCT15 cells with or without CXCL17 silence upon IL-17A supplementation; B. CCK-8 assay for cell resistance was applied to detect the cell viability of HCT15 cells with or without CXCL17 silence which treated with different concentrations of taxol; C. Wound-healing assay was used to evaluate cell migration ability of HCT15 cells treated with IL-17A followed by siNC or si-CXCL17 transfection; D. Transwell assay was applied to analyze cell invasion ability of HCT15 treated with IL-17A followed by siNC or si-CXCL17 transfection; E. Flow cytometry was used to detect the cell cycle of HCT15 cells treated with IL-17A followed by siNC or si-CXCL17 transfection; F. Flow cytometry was used to detect the cell apoptosis of HCT15 cells treated with IL-17A followed by siNC or si-CXCL17 transfection. Scale bar was 50 μm. Compared with the siNC group, **P < 0.01; compared with the si-CXCL17 group, #P < 0.05; ##P < 0.01.

Article Snippet: Drug-resistant human colorectal cancer cells (HCT15) were cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum, 1% penicillin and streptomycin, and 500 ng/ml taxol (a microtubule-stabilising drug used to treat cancer) (Med Chem Express, Monmouth Junction, NJ, USA).

Techniques: CCK-8 Assay, Wound Healing Assay, Migration, Transfection, Transwell Assay, Flow Cytometry

Journal: Cancers

Article Title: RBM47-Induced Gasdermin A/GSDMA Mediates Mesenchymal–Epithelial Transition and Pyroptosis of Colorectal Cancer Cells

doi: 10.3390/cancers18030504

Figure Lengend Snippet: RBM47 enhances chemosensitivity of CRC cells to Oxaliplatin via GSDMA. ( A ) RBM47 mRNA expression in CRC patients from the GSE19860 cohort who received FOLFOX treatment. Patients were stratified into responders (n = 9) and non-responders (n = 20) based on treatment outcome. ( B ) Representative phase-contrast images of DLD1 and HCT15 cells treated with vehicle (control), Nigericin (2 μM), Oxaliplatin (5 μM), or 5-fluorouracil (5-Fu, 8 μM) for 48 h, showing pyroptosis-specific morphological changes. Scale bars, 50 μm. ( C ) Western blot analysis of RBM47 protein expression in DLD1 cells treated with Nigericin (2 μM), Oxaliplatin (5 μM), or 5-Fu (8 μM) for 48 h. α-tubulin served as a loading control. ( D ) Correlation between RBM47 mRNA expression and Oxaliplatin IC50 values across 36 human CRC cell lines (data from CCLE and GDSC). The Pearson correlation coefficient (r) with two-tailed p -value is shown. ( E ) Cell viability curves of DLD1 cells transfected with control or RBM47 -specific siRNA and treated with increasing concentrations of Oxaliplatin for 72 h. IC50 values are indicated. ( F ) Cell viability curves of SW480/pRTR- RBM47 -VSV cells treated with or without doxycycline (DOX) for 72 h, followed by Oxaliplatin treatment at increasing concentrations for 72 h. IC50 values are indicated. ( G ) Correlation between GSDMA mRNA expression and Oxaliplatin IC50 values across 36 human CRC cell lines (data from CCLE and GDSC). The Pearson correlation coefficient (r) with two-tailed p -value is shown. ( H ) Cell viability curves of DLD1 cells transfected with control or GSDMA -specific siRNA and treated with increasing concentrations of Oxaliplatin for 72 h. IC50 values are indicated. ( I ) Cell viability curves of SW480/pRTR- GSDMA -VSV cells treated with or without DOX for 72 h, followed by Oxaliplatin treatment at increasing concentrations for 72 h. IC50 values are indicated. ( J ) Cell viability curves of SW480/pRTR- RBM47 -VSV cells transfected with control or GSDMA -specific siRNA, treated with or without DOX, and exposed to increasing concentrations of Oxaliplatin for 72 h. IC50 values are indicated. ( K ) Cell viability curves of DLD1/pRTR- GSDMA -VSV cells transfected with control or RBM47 -specific siRNA, treated with or without DOX, and exposed to increasing concentrations of Oxaliplatin for 72 h. IC50 values are indicated. Mean values ± SD (n = 3) are provided. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: SW480, SW620, DLD1, and HCT15 colorectal cancer cell lines (authenticated by STR analysis (Eurofins)) were propagated in McCoy’s 5A medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS; Invitrogen), penicillin (100 U/mL), and streptomycin (0.1 mg/mL).

Techniques: Expressing, Control, Western Blot, Two Tailed Test, Transfection

Journal: Cancer Research

Article Title: Molecular Inversion Probe Analysis of Gene Copy Alterations Reveals Distinct Categories of Colorectal Carcinoma

doi: 10.1158/0008-5472.can-06-0595

Figure Lengend Snippet: Figure 2. Heat map of cluster from significant gene copy alterations in colorectal cancer cell lines and carcinoma samples. Unsupervised hierarchical clustering was done on the data set of significant gene copy regions as determined by CLAC. The dendrogram from the clustering algorithm was used to generate a heat map, graphically depicting the relationship among the colorectal cancer cell lines. The heat map of the data uses a green (losses) versus red (gains) color scale. Each column represents an individual MIP arranged by chromosome and physical location in increasing order from left to right. A, each row represents a separate colorectal cancer cell line. The two topmost branches are designated as clusters 1 and 2. A subcluster branch representing MSI cell lines is labeled as 1a. B, each row represents a separate colorectal carcinoma sample. The two topmost clusters are labeled as clusters 1 and 2.

Article Snippet: Colorectal cancer cell lines (CACO2, COLO320, DLD1, HCT116, HCT15, LOVO, LS174T, LS180, NCIH508, NCIH747, RKO, SKCO1, SW1116, SW403, SW48, SW480, SW620, and SW837) were obtained from the American Type Culture Collection (Manassas, VA) and grown under the recommended conditions according to the manufacturer’s instructions.

Techniques: Labeling

Journal: Cancer Research

Article Title: Molecular Inversion Probe Analysis of Gene Copy Alterations Reveals Distinct Categories of Colorectal Carcinoma

doi: 10.1158/0008-5472.can-06-0595

Figure Lengend Snippet: Figure 3. Overall frequency of deletions and amplifications of clusters 1 and 2 classification of primary colorectal carcinomas. The gene copy frequency between cluster 1 and 2 is separately plotted. Y axis, overall frequency of gene copy changes for individual MIP; X axis, location of probe as arranged by chromosome and their nucleotide location.

Article Snippet: Colorectal cancer cell lines (CACO2, COLO320, DLD1, HCT116, HCT15, LOVO, LS174T, LS180, NCIH508, NCIH747, RKO, SKCO1, SW1116, SW403, SW48, SW480, SW620, and SW837) were obtained from the American Type Culture Collection (Manassas, VA) and grown under the recommended conditions according to the manufacturer’s instructions.

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Role for polo-like kinase 4 in mediation of cytokinesis

doi: 10.1073/pnas.1818820116

Figure Lengend Snippet: Western immunoblot of 19 different ovarian cell lines and one colorectal (NCI-H747) cancer cell line demonstrates full-length PLK4 ( Upper Left and Upper Right ) and phospho-PLK4 ( Lower Left and Lower Right ). ( Upper ) Western blots demonstrate full-length (97-kDa) PLK4 with a commercially available anti-PLK4 antibody (P005; Cell Signaling 3258, recognizing an epitope centered on the cysteine458 amino acid). ( Lower ) Western blots were performed by using the same filter after washing with buffer and reprobing with an anti–phospho-serine305-PLK4 antibody (Ab#14299). Please note that the phospho-PLK4 in the lower filter is of substantially smaller size, ∼60 kDa. The difference is appreciated as the full-length signals were not completely removed by stripping in the lower filters. Cultured cells were not synchronized with regard to cell cycle. We confirmed that the 60-kDa band corresponded to pPLK4 by removing this band from the gel and performing amino acid sequence analyses. We also confirmed that a second anti–phospho-PLK4 antibody (Ab#3) from our laboratory recognized phospho-PLK4 by using immunoprecipitation assays followed by PAGE with silver staining to identify the band recognized on Western blot and, finally, MS to assess the amino-acid sequence of protein band (amino-acid sequence provided in ). SI Appendix , Fig. S1 shows results in 23 additional colorectal and 9 breast cancer cell lines. ( Lower ) Schematic illustration of PLK4 amino-acid sequence shows the relative locations of epitopes recognized by the two antibodies used for Western immunoblot analyses ( Upper ), one from the laboratory of G.P.N. recognizing phospho-serine305-PLK4, produced in collaboration with Cell Signaling Technology, and the second commercially available recognizing cysteine458-PLK4.

Article Snippet: We used breast cancer (HCC1569, HCC1954, BT549, MDA-MB-415, HCC202, MCF7, T47D, CAL51), ovarian cancer (KOC-7C, SKOV3, HEYC2, COLO704, RMG-1, KK, ES-2, EFO21, MCAS, A2780, OV167, PEA2, TOV-21G, OV90, OV207, Kuramochi, OVISE, OVMANA, PEO6, CaOv-3, OV177, OVCAR3, OVCAR5, OVSAHO, OVTOKO, PEO14), colorectal cancer (HCT-116, HCT-15, DLD-1, COLO 205, COLO320DM, COLO320HSR, NCI-H747, NCI-H716, COLO201, LS1034, SNU-C2B, SNU-C1, LS513, LS411N, ATR-FLOX, LS174t, WiDr, RKO, SK-CO1, HCT-8, LS-180, HT-29, C2Bbe1, SW480), and immortalized breast epithelial (MCF-10–2A, MCF-12A) cell lines from the American Type Culture Collection (ATCC) or the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) for our experiments grown as described elsewhere ( – ).

Techniques: Western Blot, Stripping Membranes, Cell Culture, Sequencing, Immunoprecipitation, Silver Staining, Produced