The Effect Of Elemene Combined With Chemotherapy Drugs On The Proliferation Of Esophageal Cancer Cells And The Expression Of PD-L1, PTEN/PI3K/AKT Related Proteins

OBJECTIVE: To investigate the effects of elemene combined with cytotoxic drugs nedaplatin and fluorouracil on the proliferation,apoptosis,and division cycle of human well-differentiated esophageal squamous carcinoma cell line Ec109 and related proteins of PD-L1 and PTEN/PI3K/AKT pathways The changes explored the mechanism of elemene acting on human well-differentiated esophageal squamous carcinoma cell line Ec109,providing a theoretical basis for clinical application.Materials and methods: 1.CCK-8 method was used to detect and calculate the IC50 of elemene,nedaplatin,and fluorouracil,and the inhibition rate of Ec109 cell line was detected by combination therapy.2.Application of flow cytometry detection of elemene,nedaplatin,fluorouracil,elemene combined nedaplatin,elemene combined fluorouracil,nedaplatin combined fluorouracil,elemene combined nedaplatin and fluorouracil on Ec109 cells The effect of apoptosis and cycle was detected.3.The expression levels of PD-L1,PTEN,PI3 K and AKT proteins in Ec109 cell lines after treatment with various drugs were detected by Western blot.Result:The inhibitory effect of elemene(concentration?60ug/ml),nedaplatin and fluorouracil on human well-differentiated esophageal squamous carcinoma cell line Ec109 showed the time-concentration effect relationship,the difference was statistically significant(P<0.05).The IC50 s of drugs acting on cells for 48 h were: elemene 70.873 ug/ml,nedaplatin 60.358um/ml,and fluorouracil 84.159 um/ml.Follow-up experiments were conducted with elemene60 ?g/ml and nedaplatin 50 ?m/ml.Fluorouracil 50um/ml is the concentration parameter.The combination of elemene,nedaplatin,and fluorouracil can inhibit cell proliferation more effectively than single drug.The combination of elemene,nedaplatin,and fluorouracil is better than the combination of two drugs and single drug.The inhibition of cell proliferation activity was statistically significant(P<0.05).2.Elemene with low concentration(15ug/ml,30ug/ml)had a proliferative effect on Ec109 cell line(P<0.05).The absorbance OD of live cells was higher than that of blank group.Statistical significance(P<0.05).3.The combination of elemene,nedaplatin and fluorouracil can increase the rate of cell apoptosis compared with single drug,and make the cells stagnate more in S and G2 phases,and shorten the proportion of G1 phase,the difference is statistically significant(P< 0.05).The combination of three drugs of elemene,nedaplatin and fluorouracil can increase the rate of cell apoptosis compared with the two drugs and make the cells more stagnant in S and G2 phases,shortening the proportion of G1 phase,and the difference is statistically significant(P<0.05).4.The expression of PTEN protein in all combination and single drug group Ec109 cell lines showed a rising trend compared with the blank control group,the difference was statistically significant(P<0.05);all combined drug and single drug group Ec109 cell lines The expression of PD-L1,PI3 K and AKT protein showed a decreasing trend compared with the blank control group,and the difference was statistically significant(P<0.05).5.The expression of PTEN protein in Elemene plus nedaplatin group,Elemene plus fluorouracil group was lower than nedaplatin,fluorouracil single drug group and the combination of two drugs Ec109 cell line,the difference was statistically significant(P <0.05).<0.05);Compared with single agent group,the expression of PI3K/AKT protein in all combination groups was decreased,and the difference was statistically significant(P<0.05).6.The expression of PD-L1 protein in the Elemandra and Nedaplatin group,Elemene plus fluorouracil group,Elemene plus nedaplatin and fluorouracil group was lower than that of the single drug group Ec109 cell line,and the difference was statistically significant(P<0.05).<0.05).Conclusion:1.Elemene(concentration ? 60ug/ml)can inhibit the proliferation of human well-differentiated esophageal squamous cell carcinoma cell line Ec109;low concentrations(15ug/ml,30ug/ml)elemene can promote human well-differentiated esophageal squamous cell carcinoma Ec109 strain proliferation.2.Elemene combined with nedaplatin and fluorouracil can increase the inhibitory rate of human well-differentiated esophageal squamous carcinoma cell line Ec109 compared with single drug and two drugs.3.Elemene combined with nedaplatin and fluorouracil can increase the apoptosis rate of human well-differentiated esophageal squamous carcinoma cell line Ec109 and the proportion of S and G2 phase,and shorten the proportion of G1 phase.4 Elemene alone or in combination with nedaplatin and fluorouracil can inhibit the proliferation and promote apoptosis of human well-differentiated esophageal squamous carcinoma cell line Ec109.The expression of PTEN protein is increased,the expression of PI3 K and Akt protein is decreased,and the mechanism of action of the drug may be related to Negatively regulates PTEN/PI3K/Akt/ signaling pathways.5.Elemene alone or in combination with nedaplatin and fluorouracil can down-regulate PD-L1 protein expression in human well-differentiated esophageal squamous carcinoma cell line Ec109.This suggests that reducing PD-L1-mediated immune escape may be related to the mechanism of drug action in the human body.