ABSTRACT
Objective: To investigate the probable mechanism of FSEER (ethanol extract of Forsythia suspensa root)-induced apoptosis of esophageal cancer TE-13 cells in vitro . Methods: Change of mitochondrial membrane potential in TE-13 celsl after treatment with different concentrations of FSEER (0.4, 0.5 and 0.6 mg/mL) for 24 h was examined by FCM (flow cytometry). The expressions of apoptosisassociated proteins [cleaved caspase-3, caspase-8, cleaved caspase-9 and Cyt-C (cytochrome-C)] in esophageal cancer cells after FSEER treatment were detected by Western blotting. The expression levels of Bax, Bad, Noxa, Bcl-2, Bcl-xl and Mcl-1 mRNAs in TE-13 cells after treatment with different concentrations of FSEER for 24 h were detected by RT (reverse transcription)-PCR. Results: FSEER (0.4, 0.5 and 0.6 mg/mL for 24 h) can significantly reduce the mitochondrial membrane potential of TE-13 cells in a dose-dependent manner, as compared with the TE-13 cells without any treatment (P < 0.05). The result of Western blotting showed that the expression levels of cleaved caspase-3, cleaved-caspase-9 and Cyt-C in TE-13 cells after FSEER treatment were significantly increased but the expression level of caspase-8 had no change as compared with those of the TE-13 cells without any treatment (P < 0.05). RT-PCR showed that FSEER could up-regulate the expression levels of Bax, Bad and Noxa mRNAs and down-regulate the expression levels of Bcl-2, Bcl-xl and Mcl-1 mRNAs. Conclusion: The underlying mechanism of FSEER-induced apoptosis of TE-13 cells may be related to the mitochondria-dependent pathway. Copyright © 2013 by TUMOR.
ABSTRACT
Objective:To investigate the effect of ethyl acetate extract from Cortex periplocae (CPEAE) on apoptosis of human esophageal carcinoma cell line TE-13 and to elucidate its mechanism. Methods:Inhibitory effect of CPEAE on TE-13 proliferation was tested by MTT assay. The morphological changes of cell apoptosis were observed by Giemsa staining and transmission electron microscopy. Cell cycle and apoptotic ratio were tested by flow cytometry (FCM). The protein expression of CDK4 was observed by Western blotting.Results:CPEAE inhibited proliferation of TE-13 cells in a concentration-dependent and time-dependent manner, and its IC_(50) value was (2.443±0.005) μg/mL at 48 h (P<0.05). The characteristic morphological changes of apoptosis were observed in TE-13 cells after treatment with CPEAE under transmission microscope. A typical subdiploid peak was detected by flow cytometry. CPEAE decreased the expression of gene CDK4 in TE-13 cells. Conclusion:CPEAE can induce apoptosis of TE-13 cells. The effect is related with down-regulation of CDK4 expression.