Journal: PLoS ONE

Article Title: Targeting KRAS Oncogene in Colon Cancer Cells with 7-Carboxylate Indolo[3,2- b ]quinoline Tri-Alkylamine Derivatives

doi: 10.1371/journal.pone.0126891

Figure Lengend Snippet: Antiproliferative activity (IC 50 and IC 65 in μM) ^a of compounds 1a, 2a, 2d, TMPyP4 and 5-fluorouracil (5-FU) in malignant (HCT116, SW620) and non-malignant (HEK293T, CCD18co) cell lines.

Article Snippet: SW620 human colorectal adenocarcinoma cells (European Collection of Cell Cultures, ECACC, Porton Down, Wiltshire, UK), HEK293T human embryonic kidney cells (ATCC, LGC Standards, UK) HCT116 human colon carcinoma cells (ECACC), HCT116 p53(+/+) and p53(−/−) isogenic human colon cancer cells (GRCF Cell Center and Biorepository, the Johns Hopkins University, School of Medicine, Baltimore, MD, USA), were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, and 1% antibiotic/antimycotic (Invitrogen, Grand Island, NY, USA), CCD18co human colon fibroblasts (ATCC, CRL-1459) were grown in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum, 1% antibiotic/antimycotic, 2 mM GlutaMAX, 0,1 mM Non-essential amino acids (NEAA) (Invitrogen) and 0.57 mM Recombinant Human TNF-α (Peprotech, USA) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: Targeting KRAS Oncogene in Colon Cancer Cells with 7-Carboxylate Indolo[3,2- b ]quinoline Tri-Alkylamine Derivatives

doi: 10.1371/journal.pone.0126891

Figure Lengend Snippet: Cell populations were obtained by Guava ViaCount flow cytometry following 72 h incubation of HCT116 colon cancer, SW620 metastatic colon cancer, CCD18co human colon fibroblasts and HEK293T embryonic kidney cell lines with 5-FU, TMPyP4 and IQ3A at equitoxic (IC 50 and IC 65 ) concentrations, or DMSO (vehicle control). Results are expressed as the mean percentage (%) of viable, mid-apoptotic and dead cells ± SEM, of at least three different experiments, for A. IC 50 , and for B. IC 65 compound concentrations. a, p < 0.05; b, p < 0.01 from DMSO (vehicle control); c, p < 0.05; and d, p < 0.01 from 5-FU.

Article Snippet: SW620 human colorectal adenocarcinoma cells (European Collection of Cell Cultures, ECACC, Porton Down, Wiltshire, UK), HEK293T human embryonic kidney cells (ATCC, LGC Standards, UK) HCT116 human colon carcinoma cells (ECACC), HCT116 p53(+/+) and p53(−/−) isogenic human colon cancer cells (GRCF Cell Center and Biorepository, the Johns Hopkins University, School of Medicine, Baltimore, MD, USA), were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, and 1% antibiotic/antimycotic (Invitrogen, Grand Island, NY, USA), CCD18co human colon fibroblasts (ATCC, CRL-1459) were grown in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum, 1% antibiotic/antimycotic, 2 mM GlutaMAX, 0,1 mM Non-essential amino acids (NEAA) (Invitrogen) and 0.57 mM Recombinant Human TNF-α (Peprotech, USA) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Flow Cytometry, Incubation, Control

Journal: PLoS ONE

Article Title: Targeting KRAS Oncogene in Colon Cancer Cells with 7-Carboxylate Indolo[3,2- b ]quinoline Tri-Alkylamine Derivatives

doi: 10.1371/journal.pone.0126891

Figure Lengend Snippet: A. Nuclear morphology and representative images of HCT116 human colon cancer cells and CCD18co human colon fibroblasts after Hoechst staining, evaluated by fluorescence microscopy after 72 h exposure to equitoxic (IC 50 ) concentrations of 5-FU, TMPyP4 and IQ3A or DMSO (vehicle control) at 400x magnification. White Arrows indicate nuclear fragmentation and chromatin condensation, and B. Apoptotic cell populations obtained by Guava Nexin flow cytometry following 72 h incubation of HCT116 and CCD18co cells with 5-FU, TMPyP4 and IQ3A at equitoxic (IC 50 ) concentrations, or DMSO (vehicle control). Results are expressed as the mean percentage (%) of early (LR quadrant) and late (UR quadrant) apoptotic cells. Results are expressed as mean ± SEM of at least three independent experiments; *p < 0.05 and §p < 0.01 from DMSO (vehicle control); and †p <0.05 and ‡p < 0.01 from 5-FU.

Article Snippet: SW620 human colorectal adenocarcinoma cells (European Collection of Cell Cultures, ECACC, Porton Down, Wiltshire, UK), HEK293T human embryonic kidney cells (ATCC, LGC Standards, UK) HCT116 human colon carcinoma cells (ECACC), HCT116 p53(+/+) and p53(−/−) isogenic human colon cancer cells (GRCF Cell Center and Biorepository, the Johns Hopkins University, School of Medicine, Baltimore, MD, USA), were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum, and 1% antibiotic/antimycotic (Invitrogen, Grand Island, NY, USA), CCD18co human colon fibroblasts (ATCC, CRL-1459) were grown in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum, 1% antibiotic/antimycotic, 2 mM GlutaMAX, 0,1 mM Non-essential amino acids (NEAA) (Invitrogen) and 0.57 mM Recombinant Human TNF-α (Peprotech, USA) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Staining, Fluorescence, Microscopy, Control, Flow Cytometry, Incubation