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To explore the effect of arsenic on cell proliferation, the antagonism of PC against arsenic and the relationship associated to wnt/β-catenin signaling pathway by using Proanthocyanidins (PC) and sodium arsenite (NaAsO2) to intervene human hepatocyte L-02.
We treated cells with different doses of PC alone or in combination with NaAsO2. The cell activities were detected by Cell Counting Kit-8 (CCK-8) method. Cell scratch experiment and Transwell experiment were used to detect the ability of cell migration and invasion. Real-time Polymerase Chain Reaction (Real-time PCR) method and western blotting (WB) method were used to detect the expression levels of message RNA (mRNA) and protein.
NaAsO2 increased the activity of cell proliferation (1.740±0.105), enhanced the ability of migration (556.000±3.606) and invasion (439.667±8.622). NaAsO2 up-regulated the protein expression of proto-oncogene c-myc (0.739±0.012), cyclinD1 (0.943±0.002), Vascular Endothelial Growth Factor (VEGF) (1.020±0.019), apoptosis inhibitor gene survivin (0.571±0.033)and Matrix Metalloproteinase-7 (MMP-7) (0.695±0.059). NaAsO2could also up-regulate the protein and mRNA levels of pathway core moleculars wnt3a (protein: 1.044±0.010, mRNA: 1.789±0.165) and β-catenin (protein: 0.958±0.037, mRNA: 1.596±0.217). Compared with NaAsO2 group, when PC intervention was introduced, the levels of proliferation related proteins such as c-myc (0.438±0.046) decreased. The levels of mRNA and protein of wnt3a (mRNA: 1.181±0.018, protein: 0.822±0.015) and β-catenin (mRNA: 0.965±0.078, protein: 0.832±0.064) decreased. The expression of pathway inhibitors Glycogen synthase kinase-3β (GSK-3β) (0.885±0.058) and Axin (0.749±0.016) increased.
NaAsO2 induce L-02 cell proliferation, which may be due to its activation of wnt/β-catenin signaling pathway. PC can down-regulate wnt/β-catenin pathway, thereby antagonizing the cell proliferation induced by NaAsO2.
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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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