| 1. Background and objective:Pancreatic cancer (PC) is one of the most aggressive malignancy ranked as the fourth leading cause of cancer-related mortality in developed countries. However, there are usually no symptoms in the disease’s early stages, and symptoms that are specific enough to suspect pancreatic cancer typically do not start until the disease is already in an advanced stage. By the time of diagnosis the cancer has often spread to other parts of the body. The overall 5-year survival rate is less than 5%. Thus, novel effective therapeutic methods or agents for preventing and controlling this disease are urgently needed.Deguelin is a natural compound of the flavonoid family products isolated from several plantspecies, such as Derris trifoliata Lour and Mundulea sericea (Leguminosae).Several studies have already demonstrated its excellent potential to be an anticarcinogenic and antiproliferative agent against various malignant tumors including gastric carcinoma, lung carcinoma and breast carcinoma by targeting apoptosis, cell cycle arrest and anti-angiogenesis. But the relationship between deguelin and pancreatic cancer has rarely been researched so far. Therefore,in present study, we further explored the effects of deguelin as a chemotherapeutic agent against human pancreatic cancer cells and its mechanism.2. Experimental and methods:2.1 We explored the influence of deguelin on pancreatic cancer cells by CCK-8 assay and flow cytometry analysis. The change of apoptosis related proteins were examined with western blot.2.2 The effect of up-regulation of deguelin on the migration capabilities in pancreatic cancer cells was examined using wound-healing assay.2.3 Based on transwell assay and hematoxylin-eosin staining, we investigated the role of deguelin on the migration in pancreatic cancer cells.2.4We examined the expression of MMP-2 and MMP-9 at the protein level before and after deguelin interference. To further investigate whether deguelin regulate pancreatic cancer through Hedgehog signaling, we sought to examine the protein expression levels of several key members (Shh、Gli-1、PITCH-1、PITCH-2、SUFU)of this pathway.3. Results:3.1 The results of cck-8 assay showed that deguelin inhibited the growth of pancreatic cancer cell lines in a dose-and time-dependent manner.3.2Flow cytometry showed that treatment of deguelin for 48h induced apoptosis in both cell lines.the apoptosis rate peaked when the the concentration of deguelin reached 10μM. Western blot analysis showed that the expression of two apoptosis-related proteins, cleaved caspase-3 and PARP,significantly increased with the concentration of deguelin3.3 Combined the results of transwell assay and wound-healing assay., wefound that deguelin obviously decreased the migration ability of pancreatic cancer cells. 3.4 The levels of MMP-2 and MMP-9 protein were gradually decreased with the increase of drug concentration. In addition, the Hedgehog signaling proteins were significantly changed as well.4. Conclusions:4.1Deguelin was an effectiveinhibitor of pancreatic cancer cell proliferation through inducing apoptosis.4.2 Deguelin inhibitedthe invasion and migration capabilityof the two pancreatic cancer cells.4.3 Deguelin may play its role though suppressing several key members in Hedgehog signaling pathway and down-regulating the activity of MMP-2 and MMP-9. |
PDF Full Text Request