Abstract
BackgroundCancer is one of the leading cause of mortality. Even though efficient drugs are being produced to treat cancer, conventional medicines are costly and have adverse effects. As a result, alternative treatments are being tried due to their low cost and little or no adverse effects. Our previous study identified one such alternative in rice callus suspension culture (RCSC) which was more efficient than Taxol® and Etoposide, in reducing the viability of human colon and renal cancer cells in culture with minimal or no effect on a normal cell line.MethodsIn this study, we tested the effect of RCSC by studying the dynamics of lactate dehydrogenase (LDH) in lung cancer cell lines (NCI-H460 and A549), breast cancer cell lines (MDA-MB-231 and MCF-7) and colorectal cancer cell lines (SW620 and Caco-2) as well as their normal-prototypes. Complementary analysis for evaluating membrane integrity was performed by estimating LDH release in non-lysed cells and cell viability with WST-1 assay. Fluorescence microscopy with stains targeting nucleus and cell membrane as well as caspase 3/7 and Annexin V assays were performed. Real-time quantitative RT-PCR was performed to evaluate expression of 92 genes associated with molecular mechanisms of cancer in RCSC treated ling cancer cell line, NCI-H460 and its normal prototype, MRC-5. High performance liquid chromatography (HPLC) was used to collect RCSC fractions, which were evaluated on NCI-H460 for their anti-cancer activity.ResultsLower dilutions of RCSC showed maximum reduction in total LDH indicating reduced viability in majority of the cancer cell lines tested with minimal or no effect on normal cell lines compared to the control. Complementary analysis based on LDH release in non-lysed cells and WST-1 assay mostly supported total LDH results. RCSC showed the best effect on the lung non-small carcinoma cell line, NCI-H460. Fluorescence microscopy analyses suggested apoptosis as the most likely event in NCI-H460 treated with RCSC. Gene expression analysis identified significant upregulation of cJUN, NF-κB2 and ITGA2B in NCI-H460 which resulted most likely in the arrest of cell cycle progression and induction of apoptotic process. Further, HPLC-derived RCSC fractions were less effective in reducing cell viability than whole RCSC suggesting that a holistic approach of using RCSC is a better approach in inhibiting cancer cell proliferation.ConclusionsRCSC was found to be an effective anti-cancer agent on cell lines of multiple cancer types with the best effect on lung cancer cell lines. A possible mechanism for the anticancer activity of RCSC is through induction of apoptosis as observed in the lung cancer cell line, NCI-H460.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1423-3) contains supplementary material, which is available to authorized users.
Highlights
Cancer is one of the leading cause of mortality
Treated cancer cells were found to be non-proliferative under an inverted light microscope, as there was no change in the cell density compared to the untreated control, which proliferated and grew in number crowding the culture plate at 96 h
rice callus suspension culture (RCSC) was shown to be superior to Taxol® as it effectively inhibited proliferation of lung and colon cancer cells with minimal or less effect compared to normal cells in a dose and time dependent manner
Summary
Cancer is one of the leading cause of mortality. Even though efficient drugs are being produced to treat cancer, conventional medicines are costly and have adverse effects. Cancer incidence is estimated to increase every year reaching 26 million new cases and 17 million deaths per year by 2030 [2]. Eradication of cancer seems inevitable since the core cause of its pathogenesis is the rapidly changing cell machinery. The reason behind these mutations is different from one cell to another and one patient to another. With the ever-increasing incidence of cancer, the need for better and efficient drugs is a target of most cancer researchers. Since cancer cells tend to use their normal signaling and metabolic machinery to proliferate and progress, it is difficult to target cancer cells. The adverse effects due to the targeting of normal cells surrounding cancer cells render these drugs ineffective. Identifying compounds targeting cancer cells have been the main focus of researchers over the last two decades
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