The Study Of Inhabitory Effect And Its Mechanism Of 6-methylcoumarin Alone And Combine With Paclitaxel To Ec-109 Cells

Background:Recently,cancer has gradually become one of the leading causes of death.Esophagus cancer(EC)is a common digestive tract tumor with obvious regional characteristics.It has obvious regional characteristics and is affected by many factors such as diet,living habits and environment.The common histological types are esophageal squamous carcinoma(ESCC)and esophageal adenocarcinoma(EAC).In China,the main pathological type is ESCC,and the incidence of EAC in Western countries has increased more than ESCC.Early detection and early treatment are effective measures for the treatment of this disease,and early surgery has a good prognosis.However,because its incidence is relatively hidden,it is often found in the middle and later stages,so it often loses the chance of surgery.Usually,chemotherapy,radiotherapy and other means are used to control the disease,but the prognosis is poor,and the five-year survival rate keeps low.Despite this,chemotherapy is still the main treatment.Studies have shown that paclitaxel(PTX)combined with platinum in the treatment of EC is equivalent to fluorouracil(FU)combined with platinum,becoming the first-line chemotherapy recommended by the National Comprehensive Cancer Network(NCCN).For patients with poor constitution and the elderly,it is not feasible to tolerate the side effect induced mainly by platinum,an effective anti-tumor drug combined with PTX with lower side effect will benefit patients.Traditional Chinese medicine has been reported for a long time and is reliable.Objective:Study the effects of 6-methylcoumarin with/without paclitaxel on the proliferation,cell migration,gene expression and protein expression of human esophageal cancer cell line Ec-109.,and to explore the possible mechanism of the anti-cancer effect.Methods:The concentration of COU was 0,3.125,6.25,12.5,25,50 ug/ml individually;the concentration of PTX was: 0,0.25,0.5,1,2,4 ug/ml individually.(1)MTT kit is used to detect the proliferation of Ec-109 cells in control,COU,PTX and combination groups;(2)cell scratching method is occupied to detect the migration ability of Ec-109 cells in each group,(3)The expression of Cyclin-D1,EGFR,FOXO3 a and Caspase-3 in each group was detected by Real-Time PCR;(4)Cellular immunofluorescence changes of Cyclin-D1,EGFR,FOXO3 a,and Caspase-3 proteins in Ec-109 cells are detected in each group.Results(1)After drug treatment,compared with the control group: COU and PTX significantly inhibit cell proliferation of esophageal cancer Ec-109 cells,(P<0.05).The inhibitory effect of the drug combination group was enhanced,(P<0.05).Compared with the single-agent group: the inhibitory effect of the two-drug combination group was enhanced,(P<0.05).(2)Compared with the control group: the migration ability of Ec-109 cells is significantly lowered by COU And by PTX individually,(P<0.05).After the combination of two drugs the migration ability of Ec-109 cells was significantly depressed,(P<0.05).(3)Compared with the control group: There was no difference in EGFR after COU treatment(P>0.05);PTX was down-regulated(P<0.05);the gene down-regulation was more significant after the combination of the two drugs(P<0.05).There was no difference in Cyclin-D1 gene after COU treatment(P <0.05),and it was down-regulated after PTX treatment(P<0.05).The gene down-regulation was more significant after the combination of the two drugs(P<0.05).The expression of Caspase-3 gene was up-regulated after COU and PTX treatment,and there was significant difference(P<0.05).The expression of Caspase-3 gene in Ec-109 cells was significantly increased after the combination of the two drugs(P<0.05).There was no difference in FOXO3 gene after COU treatment(P >0.05);PTX was up-regulated after treatment(P<0.05);the gene up-regulation was more significant after the combination of the two drugs(P<0.05).(4)Compared with the control group: the expression of Cyclin-D1 and EGFR protein in Ec-109 cells was decreased after the combination of the two drugs,and the expression of Caspase-3 and FOXO3 a protein in Ec-109 cells after eosinophilic combination.Conclusion:Both COU and PTX have inhibitory effects on human esophageal cancer Ec-109 cells,and the combined inhibition effect of the two drugs is more obvious.This mechanism of action may be achieved by affecting gene and protein expression to inhibit cell proliferation and migration.