Yi Qi Hua Yu Jie Du Decoction To Study The Effect Of The Intervention On The Circulating Tumor Cells In Colorectal Cancer

Objective:To investigate the effects of Prescription form of Strengthening Qi and Clearing Stasis and Poison(SQCSP) of the HCT116-luc2 colorectal cancer nude mouse of transplantation tumor cells,and to reveal the intervention mechanism.Methods:1. Cultured HCT116-LUC2 cells,made nude mice models of colorectal cancer,and screened circulating tumor cells of the nude mice models of colorectal cancer by immunomagnetic beads method,then detected the expression levels of TβR I、TβRⅡ、smad2、smad3、smad4、smad6、smad7 in the circulating tumor cells by Western-blot experimental method.2.Injected the selected circulating tumor cells into nude mice through tail vein,observed the tumor formation and metastasis rate.3.Divided the HCT116-luc2 nude mice models into blank model group, SQCSP group and 5-FU group,then detected the expression levels of TβR I、TpRⅡ、smad2、 smad3、smad4、smad6、smad7 in the circulating tumor cells by Western-blot experimental method again,and detected the amount of TGF-β1 in serum of tumor bearing nude mice by ELISA method. The aim is to reveal the possible mechanism of the effect of SQCSP on the circulating tumor cells in colorectal cancer, and to provide an effective experimental basis for clinical.Results:1.The nude mice injected with the circulating tumor cells HCT116-luc2 marked by EpCAMhigh had a higher rate of tumor formation--90%, with a higher rate of metastasis:the liver metastasis rate was 60%, lung metastasis rate was 40%, and 35% of the nude mice were with both liver and lung metastasis at the same time. The nude mice injected with the circulating tumor cells HCT116-luc2 marked by EpCAMlow did not form tumors.2. Compared with the blank model group, SQCSP group could significantly reduce the expression level of TβR IN TβRⅡ、Smad3、 Smad4,meanwhile increase the expression level of Smad7, the differences between the two groups were of statistical significance. Compared with the blank model group,5-FU group could significantly reduce the expression level of Smad4,and increase the expression level of Smad7,but did not affect the expression level of TβR Ⅰ、TβRⅡ、Smad3. There was no significant difference in the expression levels of Smad2 and Smad6 in the three groups. Compared with the blank model group, SQCSP group could significantly reduce the amount of TGF-β1 in serum of tumor bearing nude mice, the difference has significant statistical significance.while there was no significant difference between 5-FU group and blank model group.Conclusions:1.The result that the nude mice injected with the circulating tumor cells HCT116-luc2 marked by EpCAMhigh had a higher rate of tumor formation, with a higher rate of metastasis and the nude mice injected with the circulating tumor cells HCT116-luc2 marked by EpCAMlow did not form tumors accord with the fact that circulating tumor cells marked by EpCAMhigu can form tumor easily. But there were still some (10%) did not form tumor, it is worth further discussion:the circulating tumor cells which were screened by EpCAMhigh may be of false positive?Therefore, The phenotypic standardization of circulating tumor cells is worthy of further study and further research.2. SQCSP can significantly reduce the expression level of TβR Ⅰ、TβR Ⅱ、Smad3、Smad4, increase the expression level of Smad7, meanwhile reduce the amount of TGF-β1 in serum of tumor bearing nude mice,but do not affect the expression levels of Smad2 and Smad6. So we can come to the conclusion that SQCSP can significantly affect the signal intensity of TGF-β/Smads pathway in the circulating tumor cells in the nude mice model of colorectal cancer.