| Esophageal carcinoma is one of the common digestive tract malignant tumors,and is characterized by high morbidity,high mortality and poor prognosis.Esophageal carcinoma is seriously harmful to human health and the quality of life.And the main therapeutic principle of esophageal carcinoma is that surgery combined with chemotherapy and radiotherapy.While most patients who have lost the surgical opportunity,are in the middle-late stage at their first visit to the hospital,cause of onset conceals and atypical early symptom of esophageal carcinoma.So chemotherapy becomes their main treatment method.However,multi-drug resistance(MDR)which is produced in chemotherapy process of malignant tumors is the main factor that leads to the treatment failure.So MDR becomes a hot research topic in recent years.In order to overcome the significant problem of MDR,scholars set out to seek new MDR reversal agent,which also is one of the focus in the field of tumor currently.Curcumin(Cur)is a kind of fat soluble phenolic pigment,which has the characteristics of low toxicity and low adverse reactions and has the pharmacological actions of anti-inflammation,anti-tumor,anti-arteriosclerosis,promoting apoptosis of tumor cells and so on.In recent years,studies have shown that curcumin can reverse the MDR of multiple drug resistant tumors.However,there are few reports on its ability of reversing the MDR of esophageal carcinoma and the mechanism remain unknown.In this research,esophageal carcinoma drug-resistant Eca-109/VCR cells was treated with the combination of small dose of curcumin and VCR,and different methods were used to study the reversal effects of curcumin on the esophageal carcinoma drug-resistant cells and then further investigate its mechanism.The effects of the proliferation of esophageal carcinoma drug-resistant cells Eca-109/VCR treated with different concentrations of VCR and different concentrations of Cur,and further selection of reverse dose,as well as the reversal effect of Cur(20 ?mol·L-1)on Eca-109/VCR cells were detected by CCK-8 assay.The apoptosis rate of Eca-109/VCR cells treated with Cur(20 ?mol·L-1)and VCR(2 mg·L-1)alone or their combination for 24 h were detected by FCM.The effect on the protein expressions of P-gp in Eca-109/VCR cells after treatment with Cur(20 ?mol·L-1)and VCR(2 mg·L-1)alone or their combination for 24 h were measured by ELISA.The relative expression level of Wnt2,?-catenin m RNA in each group were detected by real-time fluorescent quantitative PCR.The relative protein level of Wnt2,?-catenin,MMP2 and HMGB1 in every group were measured by Western Blot.It was found that the proliferation inhibitory rates of Eca-109/VCR cells were positively correlated with the concentration of curcumin,in a dose-dependent manner after treatment with various concentrations of Cur.The reverse dose 20 ?mol·L-1 was the Maximum Cur concentration with the inhibitory rate of(7.61±0.77)%(less than 10%)in Eca-109/VCR cell.After treatment with different concentrations of VCR alone or combined with Cur(20 ?mol·L-1)for 24 h,the proliferation inhibitory rates of Eca-109/VCR cell in Cur+VCR group had a significantly higher level than that in corresponding VCR group(P<0.01).The half inhibitory concentration(IC50)of VCR were 5.13 and 1.47 mg·L-1 in all VCR groups and all Cur+VCR groups in Eca-109/VCR cell respectively.And the drug-resistant reversal fold was 3.49 in Eca-109/VCR cells.After treatment with Cur(20 ?mol·L-1)and VCR(2.0 mg·L-1)alone or their combination for 24 h,the apoptosis rate of Eca-109/VCR cells in Cur+VCR group was(18.46±2.38)%,which was higher than that in VCR group(P<0.05);the protein expressions of P-gp in Cur+VCR group was 194.28±19.46,which was significantly reduced than that in VCR group(P<0.05);The relative expression level of Wnt2,?-catenin m RNA in Cur+VCR group were(36.41±2.48)% and(27.76±2.8)%,which had a significantly decreased level than that in VCR group(P<0.05);The relative protein level of Wnt2,?-catenin,MMP2 and HMGB1 in Cur+VCR group were 0.152±0.014,0.340±0.029,0.099±0.009 and 0.188±0.010,which were significantly reduced than that in VCR group(P<0.01).The results of the study demonstrated that Cur can reverse multi-drug resistance of esophageal cancer cells Eca-109/VCR,and its mechanism may be related to the down-regulation of Wnt2,?-catenin,and P-gp,which can inhibit the activity of Wnt2/?-catenin classic pathway,and related to the down-regulation of MMP2 and HMGB1 invasive protein.Reasonable use of Cur may significantly improve the chemotherapy treatment curative effect of patients with esophageal carcinoma,and is expected to provide a new clinical therapeutic approach for patients with esophageal carcinoma. |
PDF Full Text Request