Andrographolide Isolated from Andrographis paniculata Induces Cell Cycle Arrest and Mitochondrial-Mediated Apoptosis in Human Leukemic HL-60 Cells

Abstract

The in vitro cytotoxicities of the ethanol extract of Andrographis paniculata (APE) and its main diterpenoid components were evaluated in various cancer cells. APE was found to be significantly growth inhibitory to human acute myeloid leukemic HL-60 cells with an IC₅₀ value of 14.01 μg/mL after 24 h of treatment. Among the three main diterpenoids in A. paniculata, andrographolide exhibited the highest degree of cytotoxicity followed by deoxyandrographolide while neoandrographolide was the least effective. Laser confocal microscopy and gel electrophoresis studies revealed chromosomal DNA fragmentations suggesting the occurrence of apoptosis. An increase of G₀/G₁ phase cells from 51.88 % to 78.69 % was noted after andrographolide treatment for 36 h. The G₀/G₁ phase arrest and apoptosis were associated with disappearance of mitochondrial cytochrome c and increased expression of Bax but decreased expression of Bcl-2 proteins in the inhibited cells. Although the order of all these events has not been determined, it is concluded that APE and andrographolide induce cell cycle arrest and affect an intrinsic mitochondria-dependent pathway of apoptosis by regulating the expression of some pro-apoptotic markers in HL-60 cells.

Abbreviations

APE:extract of Andrographis paniculata

DSAM:deoxyandrographolide succinic acid monoester

HIV:human immunodeficiency virus

MTT:3-[4,5-dimethylthiazol-2-yl]-3,5-diphenyl-tetrazolium bromide

AO/EB:acridine orange/ethidium bromide

PBS:phosphate buffer saline

FACS:fluorescence-activated Cell Sorting system

PI:propidium iodide

SDS-PAGE:sodium dodecyl polyacrylamide gel electrophoresis

ECL:enhance dchemiluminescence

Key words

Acanthaceae - Andrographis paniculata - andrographolide - apoptosis - cell cycle - Bax - Bcl-2 family - cytochrome c

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