Marzieh Taghizadeh; Sara Soltanian; Nahid Nasibi
Abstract
The present study was conducted to determine the volatile and non-volatile fractions and the antioxidant and anti-cancer activities of ethanolic extracts of Dracocephalum polychaetum and D. kotschyi. The volatile and non-volatile fractions were investigated by gas chromatography-mass spectrometry (GC-MS) ...
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The present study was conducted to determine the volatile and non-volatile fractions and the antioxidant and anti-cancer activities of ethanolic extracts of Dracocephalum polychaetum and D. kotschyi. The volatile and non-volatile fractions were investigated by gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The cytotoxicity effect of two ethanol extracts and the major phenolic components has been evaluated on breast and colon cancer cells by the MMT assay. GC-MS of the essential oils identified about 50 compounds, and perillylaldehyde and D-limonene were the main constituents in the essential oils of the two species. Moreover, high-performance liquid chromatography- Diode array detector analysis demonstrated that the ethanolic extract of D. polychaetum and D. kotschyi were the source of phenolic compounds such as rosmarinic acid, protocatechuic acid, naringin, apigenin, syringic acid, epicatechin, chlorogenic acid, thymol, carvacrol, rutin, p-coumaric acid, gallic acid, benzoic acid, cinnamic acid, resorcinol, quercetin, salicylic acid, 4-hydroxybenzoic acid, and ferulic acid. Rosmarinic acid and thymol were the main predominant phenolic constituents in D. kotschyi and D. polychaetum ethanolic extracts. The cytotoxicity effect of D. kotschyi and D. polychaetum ethanol extracts and the major phenolic components including rosmarinic acid, thymol, apigenin, quercetin, and nariginin has been evaluated on breast and colon cancer cells by MMT assay and results indicated IC50 values in the range of 90 to 140 (µg.ml-1) after 48 hours of treatment with ethanol extracts. Among phenolic components, thymol caused the lowest cell viability and Narengin showed the lowest anti-proliferative activity. Both extracts also showed antioxidant activity using DPPH assay. The findings of this research suggest that the Dracocephalum have precious bioactive and natural compounds with significant antioxidant and in vitro anti-cancer activities.
Sara Soltanian; Mahboubeh Sheikhbahaei
Abstract
Real-time quantitative PCR (qRT-PCR) is often used as an effective experimental method for analyzing gene expression. In this method, normalization of target gene expression levels must be performed using housekeeping genes (HKGs). HKGs are used to compensate for difference between ...
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Real-time quantitative PCR (qRT-PCR) is often used as an effective experimental method for analyzing gene expression. In this method, normalization of target gene expression levels must be performed using housekeeping genes (HKGs). HKGs are used to compensate for difference between samples due to diverse quality and quality of RNAs and different reverse transcription yield. For an ideal reference gene, constant expression levels across different samples of one experiment is necessary. In the current study, expression stability of four candidate references genes including Beta actin (ACTB), glyceraldeyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine guanine phosphoribosyl transferase (HPRT1) and Beta-2-Microglobulin (β2M) following retinoic acid (RA) treatment in embryonal carcinoma NCCIT cells were evaluated.NCCIT cells were exposed to RA (10 µM) for 14 days to induce differentiation. RT-qPCR for candidate references genes was performed and normalization between untreated and RA-treated cells was performed using identical sample input amounts. Expression of OCT4, SOX2, NANOG during RA-induced differentiation was assessed by quantitative real-time PCR. RT-qPCR results indicated significant difference in expression level of GAPDH between untreated (Ct mean: 19.36667± 0.28) and RA-treated (Ct mean: 28.94± 0.18) NCCIT cells. However, transcriptional level of ACTB, HPRT and β2M remained unchanged after RA treatment. qRT-PCR analysis using ACTB, HPRT and β2M showed treatment of NCCIT cells with RA lead to significant down regulation of OCT4 (79%), NANOG (71%) and SOX2 (96%) transcript. ACTB, HPRT and β2M were recognized as valid reference genes for analysis of gene expression during RA-induced differentiation of NCCIT cells, while GAPDH was not suitable.
