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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 assay
on human colon adenocarcinoma cell line (Caco-2) with Curcumin,
Paraquat and two different combinations Curcumin and Paraquat. In
this study, different concentrations (1, 5, 10, 25, 50 and 100 μg/ml) of
Curcumin 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 stored
in cryo preservation. The apoptosis induction was determined by
measuring Annexin V activity using Annexin V apoptosis kit. In
Clonogenic assay, the cancer cells were plated in six-well plates
overnight and treated with 40 μM curcumin for a period of 3 or 6 h. After the removal of the
drug-containing medium, the cells were washed using PBS, trypsinized and plated at a low
density (2000 cells/ well in six-well plates). The colonies were stained with crystal violet and
the number of clones in a given area was counted for each condition. In both study, the higher
concentration of the curcumin gave a good results and it indicated that the secondary
bioactive compounds of curcumin have good anticancer activity.

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