The Effect Of Dihuang Guanshitong Oral Liquid On The Esophagus Carcinoma Cell Apoptosis And β-catenin And C-myc Of Wnt Pathway

Objective : To observe the inhibiting effect of Dihuang Guanshitong oral liquid (DGOL)on the human esophageal carcinoma 109 cell(Eca109) and the esophageal carcinoma cell in model rats after radiotherapy, and to study its mechanism.Methods: The human esophageal carcinoma 109 cell(Eca-109) cultured in vitro were treated by different concentration of Dihuang Guanshitong oral liquid.The MTT, TUNEL and flow cytometry (FCM) techniques were used to observe the effect of different concentration of Dihuang Guanshitong oral liquid on cell growth and cell apoptosis of Eca109 in vitro. After radiotherapy, the chemical carcinogenesis model rats were treated orally with different concentration of "Dihuang Guanshitong oral liquid" and "control drugs" for 5 weeks to explore the effect of Dihuang Guanshitong oral liquid on the esophageal carcinoma tissues of model rats . The effect of this drug on apoptosis were detected by TUNEL;on expression of P-catenin ,the key protein of Wnt, by immunohistochemistry (IHC) and Western-blot;on expression of c-myc, the target gene on the downstream of Wnt by IHC and RT-PCR.Results: â‘  Dihuang Guanshitong oral liquid can inhibit the growth of cultured esophageal carcinoma cell Eca109 in a time and concentration- dependent manner. â‘¡ Dihuang Guanshitong oral liquid can induce change of cell apoptosis with bioequivalence F of 158.35(p<0.01). The apoptotic index increase with the enhancement of drug concentration,which indicates concentration-dependence. ? Dihuang Guanshitong oral liquid can induce apoptosis of esophageal carcinoma cell in model rats after radiotherapy with bioequivalence F of 133.95(p<0.01) ,showconcentration- dependence on cell apoptosis inducement. ?After treating with Dihuang Guanshitong oral liquid, expression of P-catenin and c-myc protein in cytoplasm and cell nucleolus of every group is decreased.There is significant difference between the low dosage group and the high dosage group(p<0.05), but no significant difference with the normal group(p> 0.05).?Dihuang Guanshitong oral liquid can inhibit the P-catenin expression of esophageal carcinoma cell in model rats with bioequivalence F of 78.59(p<0.01). There is significant difference between the high dosage group and the low dosage group (p<0.05), and significant difference with liuwei group (p<0.05).Then it downregulated the transcription of c-myc with bioequivalence F of 89.67(p<0. 01). There is significant difference between the high dosage group and liuwei group (p<0.05).Conclusion: Dihuang Guanshitong oral liquid can inhibit proliferation of EcalO9 and induce apoptosis of EcalO9 in vitro. Former in vivo test approved that the apoptotic inducement of Dihuang Guanshitong oral liquid on esophageal carcinoma cell was related with increase of p53 through inhibiting gene mutation of p53 and decrease of bcl-2, c-jun and cyclinDl.In this test, the result showed that this apoptotic inducement was related to the down expression of P-catenin and c-myc protein in cytoplasm and cell nucleolus. Dihuang Guanshitong oral liquid can inhibit the expression of P-catenin and decrease the transcription of c-myc of esophageal carcinoma cells in model rats after radiotherapy , which may be one of mechanisms of the apoptotic inducement and esophageal carcinoma suppression of this drug.