| Background and aims.Liver cancer is a common malignant tumor of the digestive tract,and the death rate of liver cancer ranks third in the world in cancer-related deaths each year.Although various methods have significantly improved the survival of liver cancer patients,the efficacy of liver cancer is still not satisfactory.Studies have shown that the proliferation,invasion,metastasis,drug resistance and recurrence of liver cancer are all related to liver cancer stem cells.Therefore,the regulation of cancer stem cells and stem characteristics has become one of the hot spots in cancer research.Current studies have shown that chemotherapy and targeted therapy can lead to changes in tumor stem cells or tumor stemness,but the mechanism of Chinese medicine’s stem regulation in liver cancer remains unclear.The traditional Chinese medicine Xiaoaiping is already in the clinical medication stage.To further clarify whether Xiaoaiping is effective,this subject intends to observe in vivo whether Xiaoaiping has an inhibitory effect on liver cancer growth and whether there are changes in liver cancer dry markers Further explore the regulatory mechanism of Xiaoaiping on liver cancer stem-related pathway regulators in order to provide a theoretical basis for Xiaoaiping in the treatment of liver cancer.Methods.1.To construct Hep3 B liver cancer subcutaneous transplanted tumor model in nude mice.Subcutaneously inject a good concentration of 1 × 107 Hep3 B cells into the right abdomen of the right back of the 4-5 week-old BALB / C nude mice,and randomly divide the mice into 2 groups,each with 6 mice,by gavage Give Xiaoaiping and normal saline.2.Observe the size of transplanted tumors and the weight of mice in different groups.All mice were weighed every 2 days,tumor volume was measured using vernier calipers,and growth and weight changes were plotted.3.m RNA level detection of dry markers and dry pathways.Tumor tissues were harvested 2 weeks after administration,weighed,m RNA was extracted by Trizol method,and q CAM was used to detect changes in genes related to tumor stem markers Ep CAM,CD13,CD90,ALDH1,CD44,CD45,and all-round factors Sox2,Nanog,Oct4.4.Detection of dry markers at the protein level.The tumor tissue harvested 2 weeks later was used to extract protein by BCA method,and Western Blot was used to detect the expression of tumor stem markers Ep CAM,CD13,CD90,ALDH1,CD44,CD45,etc.and the all-round factors Sox2,Nanog,Oct4.5.Changes in key signal pathways.After extracting m RNA and protein from tumor tissues,the m RNA and protein expression of Notch signaling pathway,Wnt / β-catenin pathway,Hedgehog pathway and Hippo pathway were detected.SPSS18.0 was used for statistical analysis of all data.Key Findings.1.Nude mouse subcutaneously transplanted tumor model was successfully constructed.2.The tumor volume of the nude mice treated with Xiaoaiping was significantly smaller than that of the control group(P <0.05),and the weight of nude mice in the Xiaoaiping group was not significantly different from that of the control group.In the end,the tumors dissected out in Xiaoaiping group were all smaller than those in the control group(1.54 g ± 0.29 g / 1.15 g ± 0.31g)(P = 0.05),the difference was statistically significant.3.Xiaoaiping has different effects on the m RNA levels of dry markers and totipotent factors.Xiaoaiping down-regulated the expression of Ep CAM,CD24,CD47,Sox2,Oct4,and SALL4,and up-regulated the expression of CD13 and ALDH1.4.The results of Xiaoaiping on dry markers and totipotent factors at the protein level are consistent with the q PCR results.Xiaoaiping up-regulates ALDH1A1,but down-regulates Ep CAM,SOX2 and Oct4.5.The effect of Xiaoaiping on dry signaling pathways: no matter whether it is m RNA or protein levels,Xiaoaiping inhibits Hippo signaling pathway,Wnt signaling pathway and Hedgehog pathway.Conclusion.1.Xiaoaiping’s in vivo experiment has the effect of inhibiting tumor.2.Xiaoaiping mainly exerts an inhibitory effect on dry markers.3.Xiaoaiping may exert its anti-cancer effect through Hippo signaling pathway,Wnt signaling pathway and Hedgehog pathway-mediated liver cancer stemness. |
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