Study On The Anti-tumor Effect And Its Mechanism Of Modified Xuezheng Decoction

Purpose:To investigate the anti-tumor effects of modified Xuezheng decoction and its related mechanisms.Methods:1. To assay the effective components of water and alcoholic extracts from modified Xuezheng decoction by LC-MS.2. The proliferation inhibitory effect of modified Xuezheng decoction on different concentrations of human gastric cancer cell line SGC-7901 cultured in vitro and its correlation with time were assayed by MTT.3. The influence of modified Xuezheng decoction on gastric cancer cell line SGC-7901 apoptosis was detected by Annexin V/PI double staining fluorescence.4. The effect of modified Xuezheng decoction on the cell cycle of different concentrations of gastric cancer cell line SGC-7901 was tested by flow cytometry.5. To verify the anti-angiogenic effect of modified Xuezheng decoction through chick embryo chorioallantoic membrane technology and experiment in vitro proliferation.6. The gene expression of VEGF, mTOR, PI3K, akt of the gastric cancer cell line SGC-7901 and its correlation of concentration were assayed by RT-PCR.7. The related protein expression of the gastric cancer cell line SGC-7901 influenced by modified Xuezheng decoction and its correlation of concentration were assayed by Western Blot.8.To establish a nude mouse model with transplanted gastric tumor by subcutaneous inoculation, we can detect the tumor growth inhibition of modified Xuezheng decoction. We alculated the inhibition rate of modified Xuezheng decoction according to the xenograft tumors volume and weight. HE staining was observed in tumor tissue morphology. Immunohistochemistry was used to detect the expression of tumor tissue signaling pathway protein molecules about antiangiogenesis. Results:1. The contents of 9 major components in modified Xuezheng decoction were Paeoniflorin,Ferulaic acid,Cinnamaldehyde, Fermononetin, Curcumin,Ligustilide,Curcumol, Ursolic Acid and Oleanolic acid.The effective composition of ethanol extract was significantly higher than that of water extract.2. Modified Xuezheng decoction shows the obvious inhibitory proliferation on the gastric cancer cell line SGC-7901, and it presents the evident correlation with time and concentration.3.Modified Xuezheng decoction could significantly induce the apoptosis of the gastric cancer cell line SGC-7901.4. After 48h intervention on the gastric cancer SGC-7901 cells of different concentration of modified Xuezheng decoction, cells in G1 phase increased significantly, and the proportion of cells also increased with the increase of drug concentration, pretented a significant dose dependence.5. Modified Xuezheng decoction has an obvious anti-angiogenesis effect.6. Modified Xuezheng decoction can down-regulate the mRNA expression of VEGF, mTOR, PI3K, Akt on the gastric cancer cell line SGC-7901.7. Different concentration of modified Xuezheng decoction can down-regulate the protein expression of VEGF, mTOR, PI3K, Akt on the gastric cancer cell line SGC-7901.8. Modified Xuezheng decoction inhibits the growth of human gastric cancer SGC-7901 cells in nude mice significantly. The inhibition rates of high, medium and low dose Modified Xuezheng decoction extract were:58.98%,45.02%,27.52%. And immunohistochemistry showed the protein expression of VEGF, CD31,HIF-1?,PI3K,p-AKT, p-mTOR,COX-2 decreased in the Modified Xuezheng decoction group with the increase of the dose.Conclusions:Modified Xuezheng decoction can obviously inhibit the SGC-7901 cell proliferation. It can induce apoptosis and regulate cell cycle of human gastric cancer cell line SGC-7901. The decoction can regulate the expression of VEGF, mTOR, PI3K, Akt and other metastasis related genes or proteins in human gastric cancer cell line SGC-7901.Modified Xuezheng decoction can reduce the protein expression of VEGF,CD31,HIF-1?,PI3K,p-AKT, p-mTOR,COX-2 in human gastric cancer SGC-7901 cells transplanted subcutaneously in nude mice with the increase of the dose. In brief, we can hold that the anti-tumor effect of Modified Xuezheng decoction is closely related to the regulation of VEGF, CD31, HIF-1, PI3K/AKT/mTOR, COX-2 and other signaling pathways.