Anticancer properties of Semen Euphorbiae towards ACHN human renal adenocarcinoma cells by inducing apoptosis.

Abstract

The main objective of the present research work was to evaluate the antitumor effects of ethanol extract of Semen Euphorbiae (EESE) in ACHN human renal carcinoma cells. The effects on apoptosis induction, cell cycle phase distribution and livin protein expression were also evaluated. MTT assay was used to assess cytotoxic effects of the extract while as clonogenic assay was used to evaluate effects on colony formation tendency. Inverted phase contrast and fluorescence microscopic techniques were used to evaluate effects of EESE on cellular morphology and apoptosis. Flow cytometry using annexin V-FITC and propidium iodide were used to quantify the extent of apoptosis and also to evaluate effects on cell cycle. Results indicate that ethanol extract of Semen Euphorbiae exhibited potent cytotoxic effects in ACHN human renal cancer cells. These effects were found to be dose-dependent as well as time dependent. Clonogenic assay revealed that EESE led to dose-dependent inhibition of colony formation in these cells. EESE-treated cells also showed evident signs of alterations and deformations in cell morphology including detachment of cells from one another forming small cluster of cells. In contrast to untreated control cells, EESE-treated cells with 10, 100 and 200 µg/ml dose showed an increase in the number of cells emitting red/orange fluorescence indicating onset and execution of apoptosis. EESE extract also led to G2/M cell cycle arrest in these cells.