Rice callus suspension culture inhibits growth of cell lines of multiple cancer types and induces apoptosis in lung cancer cell lineReportar como inadecuado




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BMC Complementary and Alternative Medicine

, 16:427

Basic research

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.

KeywordsCancer cell line Rice callus culture Cell viability Apoptosis Lactate dehydrogenase Abbreviations2,4D2,4 dichlorophenoxyacetic acid

ATCCAmerican type for culture collection

CTCFCorrected total cell fluorescence

DMEMDulbecco’s minimal essential medium

EMEMEagle’s minimum essential medium

FBSFetal bovine serum

HPLCHigh performance liquid chromatography

LDHLactate dehydrogenase

MSMurashige and Skoog

PMPlant medium

RCSCRice callus suspension culture

WHOWorld Health Organization

Electronic supplementary materialThe online version of this article doi:10.1186-s12906-016-1423-3 contains supplementary material, which is available to authorized users.

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Autor: Nafeesa Rahman - Surendar Reddy Dhadi - Aparna Deshpande - Wusirika Ramakrishna

Fuente: https://link.springer.com/







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