Evaluation of Cytotoxicity and Anti-tumor Effect of Chrozophora Tinctoria and Ferula Szowitsiana’s Extracts on Lung Cancer A549 Cell-Line

Abstract

Cancer causes significant morbidity and mortality and is a major public health problem worldwide. Lung cancer has the most mortality rate among other types of cancer in the world. Plant-derived compounds have been an important source of several clinically useful anti-cancer agents. Chrozophora tinctoria and Ferula szowitsiana have been used in Iranian traditional medicines for treatment of various diseases. In the current study we aimed to assess cytotoxic and possible anti cancer effects of these plants on A549 cell line. Materials and Methods In this study, we studied the growth inhibitory and anti-tumor activity of C. tinctoria and F. szowitsiana using human lung cancer cell line A549. The cells were exposed to methanolic and hydro alcoholic extracts of C. tinctoria and methanolic extract of F. szowitsiana at different concentrations and time durations. Along with morphologic evaluation of the cultured cells, MTT assay were carried out to characterize the cytotoxicity of the extracts. According to MTT assay results F. szowitsiana was chosen to undergo molecular studies. Expression of apoptosis related genes such as bcl-2, bcl-XL, bax, p53 and Caspase-9 as well as 18S housekeeping gene was studied using reverse transcriptase-QPCR. Results Our results showed that methanolic extract of F. szowitsiana greatly inhibited A549 cell proliferation in a concentration-dependent manner. Upon treatment with extract, a clear difference was observed in cellular morphology; the cells acquired more attenuated architecture. Although we did not observe any cytotoxic effect of methanolic and hydro alcoholic extract of C. tinctoria; our MTT assay results for F. szowitsiana’s extract showed very strong cytotoxicity effect in a time and concentration dependent manner. For the concentrations less than 400 µg/ml almost no growth inhibition occurred in comparison to the untreated control after 48 and 72 hr. However, the concentrations higher than 400 µg/ml reduced the cell viability in a concentration dependent manner. Meanwhile 800 µg/ml concentration had the best inhibitory effect on proliferation of A549 lung cancer cell lines, in comparison to positive control. The RT-QPCR results also elaborated the observed results of MTT assay in the molecular level in which F. szowitsiana’s extract had altered the expression of different anti apoptosis, pro apoptosis and tumor suppressor genes. Taken together, the up-regulation of P53, Bax and caspase-9 in a time and concentration dependent manner, confirmed the hypothesis that multiple apoptotic pathways can be considered as potential mechanism of anti-proliferative action of F. szowitsiana's plant extract on human adenocarcinoma A549 cell-line. Discussion Based on the results of cellular-molecular experiment carried out in this study we could demonstrate that F. szowitsiana’s methanol extract had in vitro cytotoxic activity and inhibitory effect on the growth of human lung cancer cell line A549.As evidenced from these preliminary results, this medicinal plant and its bioactive constituents may be considered as an attractive alternatives to serve as lead compounds in drug development for human lung cancer as an adjuvant therapy. Final conclusion Collectively, our results, suggest that the F. szowitsiana’s methanol extract has a cytotoxic effect on human lung adenocarcinoma cell, A549.Although, in presence of the extract both anti-apoptotic and pro-apoptotic genes are elevated, overall balance of them has a role in cell proliferation inhibition or maybe other mechanisms apart from apoptosis are involved. However, much remains to be done before such agent could be introduced to the clinic.