| Ellagic acid is a kind of polyphenol diolactone and a dimeric derivative of Gallic acid.It has anticancer,antioxidant and anti-inflammatory effects.It is also an important natural active ingredient in tea.At present,ellagic acid has been widely reported in inhibiting the growth of cancer cells,but there are few reports on inhibiting the proliferation and apoptosis of colon cancer cells.Therefore,in this paper,cell cloning,scratch test and Western blotting were used to explore whether ellagic acid can promote apoptosis and inhibit proliferation of colon cancer SW620 cells.SW620 cells were treated with different concentrations of ellagic acid(0,12.5,50,200 μmol/L)in logarithmic phase of colon cancer cell line SW620.The inhibitory effect of ellagic acid on the proliferation of colon cancer cells was studied by MTT method.The cell morphology of SW620 cells was further observed by microscope and HE staining.The proliferation ability of single cell was verified by cell clone formation test.Cell scratch test was used to explore the ability of cell invasion and metastasis.The activity of SW620 cells was evaluated by measuring the content of MDA and SOD,and the expression of Bax,Bcl-2,Ras,PI3K,m-TOR,TNT-α,BMP,Caspase-9,p53 and P21 was detected by Western blotting.The specific results are as follows:(1)MTT results showed that the cell survival rate decreased significantly with the increase of ellagic acid concentration in the range of 12.5 ～ 200 μmol/L(P < 0.001),and the cell survival rate decreased significantly with the increase of treatment time.(2)the results of microscopic observation showed that when the concentration was 200 μmol/L,a large number of dead cells could be observed under the microscope,and the cell outline of SW620 cells became irregular with the increase of ellagic acid concentration,and the number of surviving cells decreased significantly when treated with 50 μmol/L ellagic acid by HE staining.(3)Cell cloning experiments revealed that the rate of cell clone formation was highly significantly reduced when cells were treated with 12.5 μmol/L(P <0.001),and the rate of cell clone formation was already reduced by 14.18% when the concentration reached 50 μmol/L.(4)The results of cell migration experiments showed that compared with the blank control group,the cell migration rate decreased significantly(P < 0.05)when the ellagic acid concentration was 12.5 μmol/L,and the migration rate was91.34% at this time,and when the ellagic acid concentration reached 50 μmol/L,the cell migration rate decreased extremely significantly(P < 0.001),and the migration rate was 51.62% at this time,and with the concentration up to 200μmol/L,the cell migration rate was significantly reduced to about 15%(P <0.001).(5)When the concentration was increased to 50-200 μmol/L,there was a highly significant negative correlation between MDA content and ellagic acid concentration in SW620 cells(P < 0.001);while there was a significant positive correlation between SOD content and ellagic acid concentration(P < 0.001).(6)The results of immunoblotting experiments showed that ellagic acid significantly upregulated the expression of Bax,Caspase-9,P53 and P21 proteins and significantly downregulated the expression of Bcl-2,Ras,PI3K and m-TOR proteins,which could promote the apoptosis and also inhibit the proliferation of SW620 cells compared with the blank with.Meanwhile,the expression of TNT-α was significantly up-regulated in colon cancer cells,and BMP was highly significantly up-regulated at ellagic acid concentration of 200μmol/L(P < 0.001),indicating that ellagic acid could inhibit the production of inflammatory factors in SW620 cells and promote this cell toward apoptosis.Based on the above results,it is concluded that SW620 colon cancer cells treated with different concentrations of ellagic acid can significantly inhibit cell proliferation,reduce cell viability and promote cell apoptosis. |
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