Journal: International Journal of Molecular Sciences

Article Title: Targeted Inhibition of Colorectal Carcinoma Using a Designed CEA-Binding Protein to Deliver p53 Protein and TCF/LEF Transcription Factor Decoy DNA

doi: 10.3390/ijms26209846

Figure Lengend Snippet: Delivery of purified p28-p53 protein inhibited CRC cell proliferation and xenograft tumor growth. ( A ) Confocal microscopy images showed that after delivery into HCT116 cells, the internalized p53 protein tagged with GFP and the cell-penetrating peptide p28 escaped from lysosomes and were localized in the nuclei. ( B ) Delivery of purified p28-p53 protein into HCT116 CRC cells inhibited cell proliferation, as revealed by the ethynyl deoxyuridine (EdU) incorporation assay. The error bars represent the standard error of the mean (SEM), which was determined via two-way ANOVA. ( C ) Delivery of the p28-p53 protein did not cause as much inhibition on the cell proliferation rate of NCM460 normal colonic epithelial cells as on that of HCT116 and LS174T CRC cells, as revealed by the CCK-8 cell proliferation assay. ( D ) Treatment of HCT116 cell subcutaneous xenograft tumor-bearing mice with purified p28-p53 protein decreased tumor growth. Representative images of HCT116 cell xenograft tumors at the endpoint of the experiment were shown; the experiment included the following: physiological saline ( n = 6), 5 mg of p28-p53 protein per kg of mice ( n = 6), 10 mg of p28-p53 protein per kg of mice ( n = 5), 5 mg of p28-GFP protein per kg of mice ( n = 6), 10 mg of p28-GFP protein per kg of mice ( n = 6), and 5 mg cisplatin per kg of mice ( n = 5). Administration started when the tumors grew to 70 mm 3 in volume. Statistical significance is denoted as follows: *** p < 0.001, **** p < 0.0001.

Article Snippet: The cells used in this study included (i) the human colorectal cancer cell line HCT116 (ATCC#CCL-247 TM ), with wild-type p53; (ii) the human colorectal cancer cell line SW480 (ATCC#CCL-228 TM ), with two point mutations of p53 (R273H/P309S); (iii) the human colorectal cancer cell line LS174T (ATCC#CCL-188 TM ), with wild-type p53; (iv) the normal human colonic epithelial cell line NCM460; and (v) the human cervical cancer cell line HeLa (ATCC#CCL-2 TM ).

Techniques: Purification, Confocal Microscopy, Inhibition, CCK-8 Assay, Proliferation Assay, Saline

Journal: PLOS One

Article Title: Structural and functional analysis of Escherichia coli membrane disruption by Ib-M peptides

doi: 10.1371/journal.pone.0334029

Figure Lengend Snippet: The cytotoxic activities of the peptides were measured in (A) NCM460 non-cancerous, (B) SW480, and (C) Caco2 cancer cell lines using MTT assay. Triton X-100 (1%) was utilized as a positive control for cytotoxicity. The data represent the mean ± SD of a representative assay (n = 3). The dotted line corresponds to the IC 50 value or 50% cytotoxicity, which defines the pharmacological potency of the molecule.

Article Snippet: Tissue culture cells NCM460 (normal human colon mucosal epithelial cells) ATCC CVCL-0460, SW480 ATCC CCL-228, and Caco2 (colorectal adenocarcinoma cells) ATCC® HTB-37TM were obtained from ATCC (Manassas, VA, USA) and cultured under standard conditions using Dulbecco’s Modified Eagle Medium (DMEM) high glucose (Sigma-Aldrich Co., St. Louis, MO, USA) in a humidified incubator with an atmosphere of 95% air and 5% CO 2 at 37° C. DMEM was supplemented with 10% heat-inactivated fetal bovine serum (FBS, Invitrogen Ltd, Paisley, UK), 2 mM L-glutamine, 1 mM sodium pyruvate, sodium bicarbonate, 1% non-essential amino acids, 100 units/mL penicillin, and 100 μg/mL streptomycin (Sigma-Aldrich Co., St. Louis, MO, USA).

Techniques: MTT Assay, Positive Control