Cultured human colon adenocarcinoma cells

Rutzky CP, Kaye CJ, Siciliano, MJ, Chao M Kahan BD (1980) Longitudinal karyotype and genetic signature analysis of cultured human colon adenocarcinoma cell Unes LS180 and LS174T. Cancer Research 40 1443-1448. 

The prediction of the important structural features that affect intestinal permeability is useful information to obtain early in the drug discovery process. The two most common models used to obtain fast, high-throughput measurements are the parallel artificial membrane permeation assay (PAMPA) and the cell line assays that feature cultured human colon adenocarcinoma cells (Caco-2). Each method uses a surrogate model to mimic intestinal absorption followed by LC-MS analysis. 

Cultured Human Colon Adenocarcinoma Cells (Caco-2)... 

Pinto, M., Robine-Leon, S., Appay, M.D., Kedinger, M., Triadou, N., Dussaulx, E., Lacroix, B., Simon-Assmann, P., Haffen, K., Fogh, and Zweibaum, A., Enterocytic-like differentiation and polarization of the human colon adenocarcinoma cell line Caco-2 in culture, Biol. Cell, 47,... 

These models consist of cells grown on permeable inserts. Transport of compounds across the cell monolayer can be used to quantitate the permeability of a new chemical entity in a rapid manner. One of the most popular cell lines is Caco-2, derived from human colon adenocarcinoma cells. The monolayer exhibits ion conductance and possesses transepithellal electrical resistance indicative of fully formed tight junctions that restrict the paracellular transport of a chemical entity. Although Caco-2 cells are the most commonly used cells, Madin-Darby Canine Kidney (MDCK) cells are becoming more widespread in use, in part because of the shorter culture time (4-7 days versus 21-30 days for Caco-2 cells) needed for their use in permeability experiments. 

The introduction of new technologies in recent years is increasing the throughput of ADME studies during the drug discovery process. The advent of cell culture techniques in recent years has facilitated the assessment of intestinal permeability for many drugs. A notable example is the use of CaCO-2 cells, an immortalized human colon adenocarcinoma cell line... 

Cell monolayers grown on permeable culture inserts form confluent mono-layers with barrier properties and can be used for drug absorption experiments. The most well-known cell line for the in vitro determination of intestinal drug permeability is the human colon adenocarcinoma Caco-2 [20, 21], The utility of the Caco-2 cell line is due to its spontaneous differentiation to enterocytes under conventional cell culture conditions upon reaching confluency on a porous membrane to resemble the intestinal epithelium. This cell model displays small intestinal carriers, brush borders, villous cell model, tight junctions, and high resistance [22], Caco-2 cells express active transport systems, brush border enzymes, and phase I and II enzymes [22-24], Permeability models... 

Recently, Oryza sativa L. japonica cv. Dongjin cell cultures were used to produce an antibody against the tumor-associated glycoprotein 72 (TAG 72). TAG 72 is commonly expressed in human adenocarcinoma cells. Recombinant antibody purified from transgenic rice cells was shown to bind human LS 174T colon adenocarcinoma cells expressing TAG 72. This proved the ability of rice cells to produce a functional foreign protein. 

Desmoulin, F., Galons, J.-P., Canioni, P., Marvaldi, J., Cozzone, P.J. (1986). 31P nuclear magnetic resonance study of a human colon adenocarcinoma cultured cell line. Cancer Res. 46,3768-3774. 

Peters WHM, Reolofs HMJ (1989) Time-dependent activity and expression of glutathion-S-transferases in the human adenocarcinoma cell line CACO-2. Biochem J 264 613-616 Rosenberg DW, Leff T (1993) Regulation of cytochrom P450 in cultured human colonic cells. Arch Biochem Biophys 300 186-192... 

Two new cyclic heptapeptides, scytalidamides A 43 and B 44, were isolated from the culture broth of a marine fungus, Scytalidium sp., collected from the Bahamas. Compounds 43 and 44 showed moderate in vitro cytotoxicity toward HCT-116 human colon adenocarcinoma with IC50 values of 2.7 and 11.0 pM, respectively. Both compounds displayed moderate cytotoxicity in the NCI 60 cell-line panel with mean GI50 values of 7.9 and 4.1 pM for 43 and 44, respectively. The most sensitive cell lines were MOLT-4 leukemia (3.0 pM) for 43 and Uacc-257 melanoma (1.2 pM) for 44. The total syntheses of scytalidamide A 43 was achieved on solid phase using two different resins, a phenylalanine silane resin and a 4-methoxybenzaldehyde backbone linker resin. ... 

Fig. 3. Property of gene delivery with BLs and US exposure (a) Schema of transfection mechanism by BLs and US. The mechanical effect based on the disruption of BLs by US exposure, which results in generation of some pores on plasma membrane, is associated with direct delivery of extracellular plasmid DNA into cytosol, (b) Luciferase expression in COS-7 cells transfected by BLs and US. COS-7 cells (1x10 cells/500 pLAube) were mixed wifh pCMV-Luc (5 pg) and BLs (60 pg). The cell mixture was exposed with US (Frequency 2 MHz, Duty 50%, Burst rate 2 Hz, Intensity 2.5 W/ cm. Time 10 s). The cells were washed and cultured for 2 days. Affer fhaf, luciferase acfivify was measured, (c) Effecf of US condition on transfection efficiency with BLs. COS-7 cells were exposed with US (Frequency 2 MHz, Duty 50%, Burst rate 2 Hz, Intensity 2.5 W/cm Time 0,1, 5,10 s) in the presence of pCMV-Luc (0.25 pg) and BLs (60 pg). Luciferase activity was measured as above, (d) Effect of serum on transfection efficiency of BLs. COS-7 cells in the medium containing EBS (0,10, 30, 50% (v/v)) were treated with US (Erequency 2 MHz, Duty 50%, Burst rate 2 Hz, Intensity 2.5 W/cm, Time 10 s), pCMV-Luc (0.25 pg) and BLs (60 pg) or transfected with lipoplex of pCMV-Luc (0.25 pg) and lipofectin (1.25 pg). (e) In vitro gene delivery to various types of cell using BLs and US. The method of gene delivery was same as above. S-180 mouse sarcoma cells, Colon26 mouse colon adenocarcinoma cells, B16BL6 mouse melanoma cells, Jurkat human T cell line, HUVEC human umbilical endothelial cells. Luciferase activity was measured as above. <10 RLU/mg protein, <10 RLU/mg protein Each data represents the mean S.D. n=3). L PEG-liposomes, LF Lipotectin...

The human acute lymphoblastic leukemia CEM cell line and human colon adenocarcinoma LS 174T cell line are available from the American Type Culture Collection (Rockville, MD). The human mesothelioma H-Meso cell line (32) was used previously for cytotoxicity studies (33,34). 

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