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To investigate the effect of epifriedelanol (Epi) on gene expression of P-glycoprotein (P-gp) in human colorectal adenocarcinoma cell line LS174T and its mechanism.
LS174T cells were divided into control group and experimental -L, -M, -H groups. Experimental -L, -M, -H groups were treated with 5, 10, 20 μmol·L-1 Epi, respectively. Control group was treated with 0.1% dimethyl sulfoxide. Polymerase chain reaction was used to detect the mRNA expression level of P-gp. The effect of Epi on multidrug resistance protein 1 (MDR1/P-gp) luciferase activity was investigated by pregnane X receptor (PXR)-MDR1/P-gp dual luciferase reporter gene assay. In addition, Western Blot was used to detect the protein expression level of P-gp and the nuclear factor-κB (NF-κB) pathway related proteins.
The relative expression levels of P-gp mRNA in experimental -M, -H groups and control group were 52.24±5.19, 23.00±3.52 and 100.00±9.00; the relative expression levels of P-gp protein were 86.37±9.96, 74.85±15.92 and 100.00±12.91; the relative activities P-gp luciferase were 230.19±41.32, 203.10±52.84 and 279.67±19.20; the relative expression levels of p65 (RelA/p65) in nucleus were 132.36±23.93, 145.96±25.15 and 100.00±10.88; the relative expression levels of phosphorylation NF-κB inhibits protein kinase α/β (p-IKKα/β) in cytoplasm were 184.00±54.82, 290.10±49.59 and 100.00±15.34; the relative expression levels of phosphorylated NF-κB inhibitory protein α (p-IκBα) in cytoplasm were 125.73±18.77, 133.69±20.25 and 100.00±8.12; the relative expression levels of IκBα in cytoplasm were 78.36±14.83, 70.44±14.57 and 100.00±22.82, respectively. The above indexes of experimental -M and experimental -H groups were compared with control group, and the differences were statistically significant (P<0.05, P<0.01, P<0.001).
Epi can down-regulate the gene expression of P-gp in human colorectal adenocarcinoma cell line LS174T, and the mechanism may be related to activation of NF-κB and suppression of PXR.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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