ABSTRACT
In order to study the effect and its potential mechanism of metformin combined with cisplatin treatment on human osteosarcoma MG-63 cells, MG-63 cells were treated with metformin and cisplatin and the cell proliferation and apoptosis were detected using CCK8 and flow cytometry; Cell clone formation experiment was performed to detect clone formation rate in each group; Trans well experiment was used to detect the migration and invasion ability of osteosarcoma MG-63 cells, and qPCR and Western blot were used to detect the expression of RNA and protein of apoptosis-related genes. The results showed that metformin combined with cisplatin inhibited the proliferation of human osteosarcoma MG-63 cells and promoted their apoptosis significantly (P < 0. 01); Metformin combined with cisplatin inhibited the clone formation (P < 0. 01), and the migration and invasion of human osteosarcoma MG-63 cells (P < 0. 01); Furthermore, metformin combined with cisplatin down-regulated the expression of apoptosis-related genes MCl￣1and XIAP (P <0. 01), but up-regulated the expression of apoptosis-related genes CASPASE￣3 and Cyto C (P <0. 01) and migration and invasion related genes MMP￣2 and MMP￣9 (P <0. 01). Our study indicated that metformin combined with cisplatin inhibited proliferation, promoted cell apoptosis through MCl￣1 and XIAP, and inhibited cell migration and invasion by regulating the MMP￣2 and MMP￣9 pathways in human osteosarcoma MG-63 cells.
ABSTRACT
@#[Abstract] Objective: To explore the effect of anti-ENO1 (enolase 1) antibody and metformin (MET) treatment on the proliferation, migration, invasion and stemness of cetuximab (CTX) -resistant non-small cell lung cancer (NSCLC) cells through targeting cancer stem cells and the possible mechanism. Methods: 10 mmol/L MET combined with 40 μg/ml anti-ENO1 antibody was used to treat CTX(35 µg/ml)-resistant NSCLC A549 cells for 4 d, and the effects of combined treatment on A549 cells were detected with proliferation experiment, colony formation assay, migration and invasion experiments and methylcellulose ball formation experiment. In the meanwhile, FCM was used to detect the effects of CTX, MET and anti-ENO1 antibody single-drug treatment as well as the three-drug combination treatment on ALDH+ and CD44+ lung cancer stem cell subsets. Results: CTX combined with MET and anti-ENO1 antibody treatment significantly inhibited the proliferation, migration, invasion and self-renewal capacity of A549 cells. FCM analysis found that MET could significantly inhibit ALDH+ stem cell subpopulations, while anti-ENO1 antibody could significantly inhibit CD44+ stem cell subpopulations, and the three-drug combination treatment could simultaneously suppress ALDH+ and CD44+ stem cell subpopulations. Conclusion: MET and anti-ENO1 antibody respectively target ALDH+ and CD44+ cancer stem cell subsets, and the combined treatment of MET and anti-ENO1 antibody can effectively reverse the resistance of A549 cells to CTX, and thereby more effectively inhibiting stemness, proliferation, metastasis of A549 cells and tumor recurrence.
ABSTRACT
OBJECTIVE: To investigate total glucosides paeony (TGP) on the expression phosphorylated extracellelar signal regulated kinase 1/2(p-ERK1/2), Toll-like receptors 4(TLR4) and Toll-like receptors 9(TLR9) in rats with nonalcoholic fatty liver disease (NAFLD) induced by fructose and high-fat feed. METHODS: SD rats were divided into normal group and test groups. The rats in test groups were fed with fructose and high-fat feed for 10 weeks totally to induce the test model. After 6 weeks the model was established, the rats were divided into four groups randomly, the model group (NAFLD), the metformin group (Met, 200 mg · kg-1), the low-dose TGP group(TGP-L, 100 mg · kg-1) and the high-dose TGP group(TGP-H, 200 mg · kg-1) (n=10). Four weeks later all the rats were killed and checked the indexes such as serum fasting blood glucose(FBG), fasting insulin(Fins), insulin sensitivity index (ISI), total cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), triglyceride (TG), free fatty acids (FFA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutathione S-transferase (GST) and liver index. The expression p-ERK1/2, TLR4 and TLR9 were inspected by Western-blot. RESULTS: Compared with the normal group, the rats in test groups were with the high levels serum FBG, insulin, TC, LDL-C, TG, FFA, ALT, AST, GST, liver index, p-ERK1/2, TLR4 and TLR9 (P 0.05). CONCLUSION: By downregulating the expression ERK1/2, TLR4 and TRL9, TGP can improve abnormal glucose and lipid metabolism and insulin resistance, enhance insulin sensitivity and ameliorate liver function in rats with NAFLD induced by fructose and high-fat feed.