Abstract

CLONOGENIC AND APOPTOSIS SCREENING OF CURCUMIN (CURCUMA LONGA) ON PARAQUAT INDUCED TOXICITY IN HUMAN COLORECTAL ADENOCARCINOMA [NCI-PBCF-HTB37 (CACO-2)] CANCER CELL LINE

Dilla Jose and P. Senthilkumaar

Abstract

The present study focused on the colony forming and apoptosis assayon human colon adenocarcinoma cell line (Caco-2) with Curcumin,Paraquat and two different combinations Curcumin and Paraquat. Inthis study, different concentrations (1, 5, 10, 25, 50 and 100 μg/ml) ofCurcumin and Paraquat were used. The test cancer lines (NCI-PBCFHTB37(Caco-2) - Human Colorectal Adenocarcinoma, (ATCC®HTB-37™) were procured from American Type Culture Collection(ATCC) and were subculture with suitable culture medium and storedin cryo preservation. The apoptosis induction was determined bymeasuring Annexin V activity using Annexin V apoptosis kit. InClonogenic assay, the cancer cells were plated in six-well platesovernight and treated with 40 μM curcumin for a period of 3 or 6 h. After the removal of thedrug-containing medium, the cells were washed using PBS, trypsinized and plated at a lowdensity (2000 cells/ well in six-well plates). The colonies were stained with crystal violet andthe number of clones in a given area was counted for each condition. In both study, the higherconcentration of the curcumin gave a good results and it indicated that the secondarybioactive compounds of curcumin have good anticancer activity.

Keywords: .