ABSTRACT
OBJECTIVE: To evaluate the effects of mistletoe alkali on human Osteosarcoma cells (U2OS) in vitro. SUMMARY OF BACKGROUND DATA: Osteosarcoma is the most common primary malignant tumor of bone tumor, although there are a lot of therapies such as surgery, radiotherapy and chemotherapy, its prognosis is still very poor. There is increasing interest in the protective biological function of natural antioxidants contained in Chinese medicinal herbs, which are candidates for the prevention of tumors. Mistletoe alkali is one of the compounds extracted from Viscum coloratum (Komar.) Nakai, one kind of mistletoe, whose extracts contribute to the improvement of the prognosis of patients with malignancies. METHODS: The effect of mistletoe alkali on the growth of U2OS cells was compared with 5-FU, using a Cell Counting Kit-8 (CCK-8). The influence of mistletoe alkali on U2OS's proliferation and apoptosis were tested by TUNEL staining and immunocytochemical (ICC) staining of caspase 3 and proliferating cell nuclear antigen (PCNA). Additionally, the invasion ability of U2OS cells was detected using a Boyden chamber trans-well migration assay. RESULTS: CCK-8 assays gave an IC50 of 7λg/ml for mistletoe alkali. Compared to 5-FU, mistletoe alkali inhibited U2OS proliferation and induced apoptosis more effectively. The invasion ability of U2OS was also weaker in mistletoe alkali than in 5-FU. CONCLUSIONS: Mistletoe alkali significantly inhibited growth and invasion abilities of U2OS cells and induced their apoptosis in vitro. Mistletoe alkali may be a more effective drug for Human Osteosarcoma than the standard chemotherapeutic drug 5-FU.
ABSTRACT
Objective To investigate the signal transduction pathway mechanisms of rats with liver fibrosis regulated by leptin and interfering effects of mistletoe alkali .Methods The hepatic fibrosis in rats model was established by injecting carbon tetrachloride .Forty-five SD rats were randomly divided into normal group ,model group and therapeutic group.All rats except rats in normal group were intraperitoneally injected with 40%carbon tetrachloride in peanut oil with a dose of 2.0 mL/100g according to the body weight twice a week for 8 weeks.Then, the therapeutic group was given mistletoe alkali (8g/(kg· d)) for 8 weeks via gastrogavage.Rats in normal and model group were served with distilled water at the same time.At the end of the 16th week, blood and tissue specimens were taken from all the rats .The influence of mistletoe alkali on liver morphology in liver fibrosis rat model was reviewed by HE and Masson staining .The effects of mistletoe alkali on the expression of Leptin and its receptor ( OB-Rb ) in HSC in fibrosis rat model were determined by immunohistochemistry (IH).The expression of JAK2, STAT3 and the activity of phospho -JAK2, phospho-STAT3 were detected by Western blotting analysis .Results The degree of fibrosis of the model group was more severe than the normal group and the treatment group , which suggested that mistletoe alkali can reverse liver fibrosis in rats . Immunohistochemical staining showed that mistletoe alkali reduced the hepatic expression of leptin and OB -Rb in rats with liver fibrosis in comparison with their expression in the model group .Compared with the normal group , the expression of JAK2 and STAT3 increased in the model group .However, the expression of JAK2 and STAT3 decreased in the medication groups compared with the model group .Conclusion Mistletoe alkali can effectively ameliorate liver fibrosis in rats possibly through inhibiting hepatic leptin and its receptor expressions , which through inhibiting hepatic leptin and its receptor expressions , thus inhibit the JAK2/STAT3 signaling pathway .
