HT-29 cells

H Fischer, B Illek, PA Negulescu, W Clauss, TE Machen. (1992). Carbachol-acti-vated calcium entry into HT-29 cells is regulated by both membrane potential and cell volume. Proc Natl Acad Sci USA 89 1438-1442. 

In MDR cells, a significant fraction of P-gp is found associated with caveolin-rich membranes, and there is a substantial increase in the number of caveolae and caveolin-1 protein level. For example, both multidrug resistant human colon adenocarcinoma HT-29 cells and adriamycin-resistant breast adenocarcinoma MCF-7 cells display about a 12-fold increase in caveolin expression, which correlates with an approximate fivefold increase in morphologically identifiable caveolae [55], In addition, these cells exhibit increased amounts of phospholipase D and lipids such as cholesterol, glucosylceramide, and sphingomyelin [56, 57], Similarly, taxol-resistant A549 cells display both increased caveolin expression and caveolae numbers [58], While these correlations track with MDR, they do not suggest a simple mechanism for the role of... 

Both cyclo(Gly-Leu) and cyclo(Gly-Ile) at 10 and lOOpmolG exhibited moderate effects in inhibiting HT-29, MCF-7, and HeLa cancer cell lines, except for cyclo(Gly-Ile) that showed very little activity against HT-29 cells at a concentration of 10 pmol The greatest activity was noted for 100 pmol cyclo(Gly-Leu) against HeLa cells (28.64 1.51% inhibition, P= 0.0018). °... 

Davis, C.D. and Uthus, E.O. (2002) Dietary selenite and azadeoxycytidine treatments affect dimefhylhydrazine-induced aberrant crypt formation in rat colon and DNA mefhylation in HT-29 cells. The Journal of Nutrition, 132, 292-297. 

Moved] Cranberry fruit of Early Black cultivar was fractionated chromatographically and fractions were analyzed for flavonoid content. The effects of the flavonoid fractions and ursolic acid, an abundant triterpenoid in cranberry peel, were assessed in two models of colon cancer and one model of breast cancer. Clonogenic soft agar assays were used to determine the effect of these compounds on tumor colony formation in HCT-116, HT-29 and MCF-7 cells. MTT and trypan blue assays were performed to assess their ability to inhibit tumor cell proliferation. TUNEL assays were performed to assess apop-totic response to the cranberry compounds. The proanthocyanidins inhibited tumor colony formation in HCT-116 and HT-29 cells in a dose-dependent manner, with greater effect on the HCT-116 cell line. Ursolic acid strongly inhibited tumor colony formation in both colon cell lines. These compounds also decreased proliferation in all three tumor cell lines with the HCT-116 cell line most strongly affected. (150 words)... 

Fig. 3. Effect of gemcitabine on the sensitivity of HT-29 cells to radiation. Cells were incubated with no drug, 3 nM, 10 nM, and 30 nM gemcitabine for 24 h followed by radiation. Surviving fraction was corrected for cell survival in the absence of radiation. From Shewach et al. Cancer Res 1994 54 3218-3223.

Assessment of the virulence of Listeria monocytogenes Agreement between a plaque-forming assay with HT-29 cells and infection of immunocompetent mice. Int. ]. Food Microbiol. 68, 33-44. 

Cell Lines. The human fetal intestinal cell line (FHS) was kindly supplied to us by Dr. Walter A. Nelson-Rees at the Naval Bioscience Laboratory, Oakland, California. The human colonic cell line, SKCO-1 developed by Drs. G. Trempe and L.F. Olds, was obtained from Dr. Jorgen Fogh, Sloan Kettering Institute, Rye, New York. The HT-29 cell line was developed and obtained from Dr. J. Fogh. SW-480 and SW-620 were developed at Scott and White Clinic in Temple, Texas and were obtained from Col. A. Liebovitz. 

Grow the HT-29 cell line in T75 flasks in McCoy s 5A medium (10% FBS). When cells are confluent, harvest HT-29 cells by adding 2.0 ml of TrypLE Express. After the cells detach from the flasks, resuspend the cells in McCoy s medium (10% FBS) to inactivate the TrypLE Express. Count the cell concentration as described below. Prepare two small T25 flasks with McCoy s medium (10% FBS) and seed 1 x 106 cells per flask. Keep the HT-29 cells in culture as they will be needed again later. 

Selection of an HT-29 Cell Line Stably Transduced with the Luclferase Gene... 

The next day, infect the HT-29 cells with luciferase or mock retroviral particles. A 2.5 ml solution containing retroviral particles is either prepared fresh or obtained from a frozen aliquot (see above). Prepare polybrene as 100 x stock solution (0.8 mg/ml) in 1 x PBS (can be stored at —20°C). Add polybrene to the tubes containing the retroviral particles at a final concentration of 8 pg/ml. Aspirate the medium from the HT-29 cells in the two T25 flasks. Initiate the infection by adding 2.5 ml of the solutions containing the luciferase or mock retroviral particles with polybrene to both flasks. 

One day after passaging the infected HT-29 cells onto new plates, start the selection process by adding geneticin direcdy to the medium on all four plates at a final concentration of 800 pg/ml. 

Luciferase-infected HT-29 cells start to form small colonies after 10 days of selection with geneticin. Mock-infected HT-29 cells (negative control) start to detach from the plates after about 4 days and completely disappear after 10 days in the presence of geneticin. 

FIGURE 6 Global Foodomics strategy used to investigate the activity of rosemary polyphenols against colon cancer HT-29 cells proliferation at molecular level. Reproduced from Ref. (20). 

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