Induction of HT-29 Colon Cancer Cells Apoptosis by Pyrogallol with Growth Inhibiting Efficacy Against Drug Resistant Helicobacter pylori.

BACKGROUND

Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take lead to alleviate the existing clinical challenges.

OBJECTIVE

To investigate anti bacterial activity against Helicobacter Pylori and in vitro anti colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol.

METHODS

Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cyto-toxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation was assessed with AO/ethidium bromide and DAPI staining. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay.

RESULTS

ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62% respectively. MTT results shows that HT-29 cells are more sensitive to pyrogallol with an IC50 value of 35µg/ml compare to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol induced apoptosis by simultaneous down regulation of Bcl-2 and up regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked anti metastatic potential by inhibiting the migration of HT-29 cells dose dependently.

CONCLUSIONS

Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti colon cancer agent.