Pan, Li-Li
(Department of Biochemistry and Molecular Biology, Guangxi Medical University)
,
Wang, Ai-Yan
(Department of Biochemistry and Molecular Biology, Guangxi Medical University)
,
Huang, Yong-Qi
(Department of Biochemistry and Molecular Biology, Guangxi Medical University)
,
Luo, Yu
(Department of Biochemistry and Molecular Biology, Guangxi Medical University)
,
Ling, Min
(Department of Biochemistry and Molecular Biology, Guangxi Medical University)
To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell...
To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell cycle and apoptosis with measurement of mRNA and protein levels of two apoptosis-related genes, Bcl-2 and Bax. Flow cytometry assays showed mangiferin could inhibit CNE2 cell proliferation via G2/M arrest and induction of early apoptosis. Real time PCR and Western blotting showed the mRNA and protein level of Bcl-2 to be down-regulated, while those of Bax were upregulated, when CNE2 cells were treated with mangiferin. This investigation indicated anti-proliferation effects of mangiferin through induction of cell apoptosis regulated by Bcl-2 and Bax expression.
To investigate the anti-proliferative mechanism of mangiferin in a human nasopharyngeal carcinoma cell line, CNE2 cells were incubated with different concentrations of mangiferin (12.5, 25, 50, 100, 150 and $200{\mu}M$) or with PBS as a control for 72 hours. Analyses were made of the cell cycle and apoptosis with measurement of mRNA and protein levels of two apoptosis-related genes, Bcl-2 and Bax. Flow cytometry assays showed mangiferin could inhibit CNE2 cell proliferation via G2/M arrest and induction of early apoptosis. Real time PCR and Western blotting showed the mRNA and protein level of Bcl-2 to be down-regulated, while those of Bax were upregulated, when CNE2 cells were treated with mangiferin. This investigation indicated anti-proliferation effects of mangiferin through induction of cell apoptosis regulated by Bcl-2 and Bax expression.
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문제 정의
However, the molecular mechanisms of the anti-cancer effect have not been fully elucidated. In this study, we aim to investigate the inhibitory effects of mangiferin in CNE2 cells and to explore the possible underlying mechanisms.
제안 방법
Table 1 shows the primers used in this study. After adding fluorescent probes, real-time PCR was done using a Mastercycler ep realplex real time PCR meter (Eppendorf, Germany) with 40 cycles of denaturation at 95℃ for 30 seconds, annealing at 58℃ for 30 seconds, and extension at 68℃ for 30 seconds. Relative transcription levels were calculated with the following formula: Relative quantitation = 2-△△Ct (△△Ct = (Ct mangiferin-treated group-Ct control group) Destination gene - (Ct mangiferin-treated group-Ct control group) GAPDH).
Propidium iodide (PI) (Beckman Coulter, USA) was added on ice in dark for at least 15 minutes. All samples were then analyzed on a FACS Calibur flow cytometer (Beckman Coulter Inc. USA) to determine the cell apoptosis rate and the cell number at each cell cycle.
대상 데이터
Then stock solution at 200 μM was prepared for analysis. Human nasopharyngeal carcinoma cell line CNE2 was obtained from Guangxi Medical University, China. Cells were cultured in RPMI1640 medium (Gibco, USA) supplemented with 10% fetal calf serum (Gibco, USA), 100 μg/ml penicillin (Gibco, USA), and 100 U/ml streptomycin (Gibco, USA) at 37℃ in an atmosphere containing 5% CO2.
데이터처리
The statistical analysis involving two groups was made by Student’s t test.
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