| Purpose:To study the effects of ethanol extracts of Euphorbia euphorbiae and Eucidium combined with gemcitabine on the proliferation,apoptosis and cycle of human breast cancer MDA-MB-231 cells.Material and method: 1.Take human triple negative breast cancer cell MDA-MB-231 as the research object.2.The CCK-8 method was used to detect the ethanol extracts of Euphorbia lupus: 1?g / m L,5?g / m L,10?g / m L,20?g / m L,40?g / m L,60?g / m L,80?g / m L,160?g / m L for breast cancer The role of MDA-MB-231,and calculate the inhibition rate and IC50.The effect of gemcitabine 0.25?g / m L,0.5?g / m L,1?g / m L,2?g / m L,4?g / m L,8?g / m L on breast cancer cell MDA-MB-231 was measured by CCK-8 method,and the inhibition rate and IC50 were calculated.The CCK-8 method was used to detect whether the ethanol extract of Euphorbia lupus: 1?g / m L,5?g / m L,10?g / m L,20?g / m L,40?g / m L,80?g / m L,160?g / m L had a normal human breast cell MCF-10 A Cytotoxicity,resulting in non-cytotoxic concentration.The CCK-8 method was used to detect the non-cytotoxic concentration of Euphorbia lupus ethanol extract combined with 0.25?g / m L,0.5?g / m L,1?g / m L,2?g / m L,4?g / m L,8?g / m L concentration of gemcitabine after 48 h,The effect on breast cancer cells MDA-MB-231,and calculate the inhibition rate and IC50.3.Flow cytometry to detect the cytotoxic concentration of Euphorbia euphorbiae ethanol extract without cytotoxic concentration and 48 h gemcitabine IC50 concentration alone or in combination on the breast Cancer cell MDA-MB-231 cell cycle and apoptosis.triple negative breast cancer MDA-MB-231 cells were used as the research object.2.Detection of 1 ?g / m L,5 ?g / m L,10 ?g / m L,20 ?g / m L,40 ?g / m L,60 ?g / m L,80 ?g / m L,160 ?g / m L of breast cancer MDA by CCK-8 method-MB-231 cells,and calculated the inhibition rate and IC50.The effects of gemcitabine 0.25 ?g / m L,0.5 ?g / m L,1 ?g / m L,2 ?g / m L,4 ?g / m L,and 8 ?g / m L on breast cancer MDA-MB-231 cells were measured by the CCK-8 method,and the inhibition rate and IC50.The CCK-8 method was used to detect whether 1 ?g / m L,5 ?g / m L,10 ?g / m L,20 ?g / m L,40 ?g / m L,80 ?g / m L,160 ?g / m L had normal human breast MCF-10 A cells.Cytotoxicity results in a non-cytotoxic concentration.CCK-8 method was used to detect non-cytotoxic concentrations of ethanol extracts of Euphorbia euphorbiae euphorbiae,combined with 0.25 ?g / m L,0.5 ?g / m L,1 ?g / m L,2 ?g / m L,4 ?g / m L,and 8 ?g / m L concentrations of gemcitabine for 48 h.After that,the effect on breast cancer MDA-MB-231 cells was calculated,and the inhibition rate and IC50 were calculated.3.Flow cytometry was used to detect the effects of non-toxic concentration of ethanol extract of Euphorbia euphorbiae and Euphorbia fulvae and IC50 concentration of 48 h gemcitabine on breast cancer MDA-MB-231 cell cycle and apoptosis.Results: 1.The ethanol extract of Euphorbia sibiricum acts on human breast cancer cells MDA-MB-231 alone,and the IC50 values of 24 h and 48 h are 50.389?g / m L and 31.027?g / m L respectively;Gemcitabine acts alone on human breast cancer cells MDA-MB-The IC50 values of 231,24 h and 48 h were 2.231?g / m L and 1.434?g / m L,respectively;the ethanol extract of Euphorbia lupus spp.Acting alone on normal human breast cells MCF-10 A,the concentration of 1?g / m L and 5?g / m L had no obvious cytotoxic;The ethanol extract of Euphorbia lupus,1?g / m L,5?g / m L,respectively,combined with gemcitabine on human breast cancer cells MDA-MB-231,the IC50 values of 48 h were 1.135?g / m L,0.769?g / m L,respectively.2.Euphorbia lupus ethanol extract 1?g / m L and 5?g / m L,Gemcitabine ethanol extract 1?g / m L,5?g / m L combined with gemcitabine 48 h IC50 1.4?g / m L in human breast cancer Cell MDA-MB-231,after 48 h,the cell cycle cycle arrest effect of the combined group was better than that of the gemcitabine group,and the ethanol extract of Euphorbia sibiricum can enhance the cycle arrest ability of gemcitabine.3.The ethanol extract of Euphorbia lupus,1?g / m L,5?g / m L combined with gemcitabine 48 h IC50 1.4?g / m L,respectively,can promote the apoptosis of human breast cancer cell MDA-MB-231.The apoptosis rate of Euphorbia ethanol extract combined with gemcitabine on human breast cancer cell MDA-MB-231 cells also increased significantly.Conclusion: 1.The ethanol extract of Euphorbia lupus and gemcitabine alone can inhibit the proliferation of human breast cancer cell MDA-MB-231.And within a certain range,it has the characteristics of positive correlation with time and concentration,that is,with the extension of time and the increase of drug concentration,the effect of proliferation inhibition is enhanced.When the ethanol extract of Euphorbia sibiricum is used in combination with gemcitabine,the two drugs have a synergistic effect,and the proliferation inhibitory effect on MDA-MB-231 triple negative breast cancer cells is significantly enhanced and is superior to the two drugs alone.2.After 48 hours of the ethanol extract of Stellera chamaejasme and gemcitabine on human breast cancer cells MDA-MB-231,1?g / m L and 5?g / m L of the ethanol extract of Stipa australis can enhance the cell cycle arrest effect of gemcitabine,and Its blocking effect is positively correlated with concentration.3.After 48 hours,the ethanol extract of Euphorbia lupus and gemcitabine acted on human breast cancer cells MDA-MB-231 alone.Compared with the control group,the cell apoptosis rate increased significantly.When the ethanol extract of Euphorbia sibiricum combined with gemcitabine acted on human breast cancer cell MDA-MB-231 after 48 h,the apoptosis rate was significantly increased compared with the control group,and the two had a synergistic effect,which was higher than the single drug effect.. |
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