Objective:
The present work tested organic solvents to prepare an extract with anticancer properties from a polyherbal mixture containing Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizomes). We evaluate anticancer effects in non-small-cell lung cancer cells (NCI-H292), and discuss optimization for pharmaceutical use in the context of efficacy, yield and toxicity.
Methods:
Using different organic solvents, six extracts were prepared from the polyherbal mixture. Based on the cytotoxic effects of these extracts on NCI-H292 cells and normal lung cells (MRC-5), as evaluated by the sulphorhodamine B assay, the total ethyl acetate (T-EA) extract was selected for further analysis. The possible anticancer mechanisms were assessed by evaluating the extract’s effects on apoptosis (through fluorescent microscopic analysis, DNA fragmentation analysis, caspase 3/7 assay and analysis of expression levels of apoptosis-related genes p53, Bax, survivin, Hsp70 and Hsp90), colony formation and antioxidant activity.
Results:
The extract had cytotoxic effects against NCI-H292 cells in a time- and dose-dependent manner. Significant antioxidant activity and inhibition of colony formation were also observed. The expression level of caspase 3/7 significantly (P ? 0.001) increased in NCI-H292 cells treated with 50 μg/mL of the extract. The same dosage led to a significant increase in expression levels of Bax and p53 (P ? 0.05 and P ? 0.01 respectively), accompanied by a significant decrease (P ? 0.0001) in survivin, Hsp 70 and Hsp 90.
Conclusion:
T-EA extract of the above polyherbal mixture has cytotoxicity against NCI-H292 cells via induction of apoptosis, antioxidant effects and inhibition of colony formation

Key words: Polyherbal mixture, Lung cancer, Nigella sativa, Hemidesmus indicus, Smilax glabra, Anticancer activity, Cytotoxicity, Apoptosis, Antioxidant activity