Experimental Reaserch On The Mechanism Of Proliferation And Metastasis Of Orthotopic Transplantation Human Gastric Carcinoma Of NCI-N87 Effected By Yiqi Bushen Formula

Objective:To study and explore the effect and the potential mechanism of Yiqi Bushen Formula on the growth and metastasis of orthotopic transplantation human gastric carcinoma of NCI-N87 in nude mice, which will provide theoretical evidence for clinical application.Methods:①NCI-N87 gastric cancer cell line was injected subcutaneously on 10 nude mice. when the tumor increased to the range of 1-2 cm, tumors were exposed and inoculated in nude greater curvature of the stomach serosa to establish a tumor-burdened nude mice model.②Except for Randomly extraction 55 mice were to establish the nude mice of Human gastric carcinoma constructed using orthotopic transplantation and an enrichment in cancer stem cell with 5-fluorouracil. Other 10 nude mice were set as negative control group, After two weeks, five mice in both groups were used to validate the model by immunohistochemical method. After the modeling, they were randomly divided into 4 groups:10 nude mice in saline control group,10 nude mice in 5-fluorouracil (5-Fu) group, 10 nude mice in Yiqi Bushen Formula group,10 nude mice in combination group of Yiqi bushen Formula and 5-fluorouracil, the other 5 mice were used as negative control group. Nude were intervened with drug for four weeks.③Body weight and tumor volum were measured during the treatment.④Detecting the expression level of CD24, CD44, EpCAM in tumor tissues by immunohistochemical method. Detecting content of TGF-betal in serum from different groups by ELISA.⑤Detecting content of TGF-betal in serum from different groups by ELISA.Results:①Changes of nude mice body weight. The average weight of 5-Fu group, Yiqi Bushen Formula group and combination group became significantly higher than those in saline control group (P<0.05). And there were statistical significant differences about average weight between Yiqi Bushen Formula group and the other two treatment groups (P<0.05). ②Metastasis. There was no significant metastasis focus in other organs of the nude mice.③Tumor inhibition rate. The mean of the size of tumor in three treatment groups were significantly lower than that in the saline control group (P<0.05). The anti-tumor rate was 42.23% in combination group of Yiqi bushen Formula and 5-fluorouracil, while 39.47% 5-Fu group and 33.45% in Yiqi Bushen Formula group.④IHC. The expression of CD24 and CD44 in Yiqi Bushen Formula group and combination group were found to be lower than those in saline control group and 5-Fu group (P<0.05). The positive EpCAM expression rate of Yiqi Bushen Formula group was apparently lower than saline control group and 5-Fu group (P<0.05).⑤Serum level of TGF-betal. The expression of TGF-betal in Yiqi Bushen Formula group,5-Fu group and combination group were found to be lower than those in saline control group (P<0.05).Conclusion:①Yiqi Bushen Formula could effectively inhibit the proliferation and metastasis of cancer cells, lessen the losses of body weight, improving the quality of life and realizing the state of "Survival with tumor".②Yiqi Bushen Formula could effectively inhibit the proliferation and metastasis of cancer cells which may be associate with the down-regulation the expression of CD24, CD44, EpCAM in nude mice model, reducing the proportion of the gastric cancer stem cells surface markers in vivo and inducing benign differentiation of gastric cancer stem cell. ③Yiqi Bushen Formula could effectively inhibit the proliferation and metastasis of cancer cells which may be associate with the down-regulation the expression of TGF-betal.