| Pancreatic cancer is the common digestive carcinoma, with the high malignancy. Current treatment for pancreatic cancer is unsatisfactory, which the patients hold the high mobility, and the relative survival rate in5years is only5%. The chemotherapeutics will induce the multiple side effects and the curative effect is limited. Therefore, it will bring the new potential for pancreatic cancer patients to find out one new chemotherapeutic drug or scheme.The treatment of interferon combining with other chemotherapeutics can improve the5-year survival rate of pancreatic cancer patients, and alleviate the progression of pancreatic cancer, but the application of interferon is limited. Recombinant antitumor antivirus protein (RAAP; Trade name:Novaferon) is a kind of protein engineering drug derived from interferon, its anti-tumor efficacy has been established in skin, liver, lung, and prostate cancers. The antitumor activity of RAAP is the200times of natural human interferon, and the antivirus activity is above the10times. However, the effect of RAAP on pancreatic cancer cells and the related mechanism remain unknown. To investigate the effect of RAAP on pancreatic cancer cells will supply the confirmative evidences for the clinical application of RAAP in the treatment of pancreatic cancer.The first chapter used pancreatic cancer cell line BxPC-3, to observe the effect of RAAP in different concentrations and different time periods on proliferation, apoptosis and invasion of BxPC-3cells by MTT assay, flow cytometry, fluorescence staining, and Transwell assay. The results showed that RAAP in5ng/ml concentration could inhibit the proliferation of BxPC-3cells obviously, the time period of24hours was a positive time point. Moreover, with the increment of concentration of RAAP, the inhibitive effect was more significant, as well as the cell cycle arrest in G1stage, and the apoptotic rates of cells. On the contrary, the cell invasion was alleviated with the increment of RAAP concentration. All effects of RAAP on BxPC-3cells were dose dependent.In order to explore the molecular mechanism of RAAP inhibiting proliferation and invasion of BxPC-3cells, and promoting apoptosis of BxPC-3cells, the second chapter applied RT-PCR and Western blot to determine the expressions of some key genes during cell apoptosis, angiogenesis and cell invasion. The results showed that RAAP could increase the expressions of caspase-3and caspase-8, decrease the expressions of VEGF-A and MMP-2, while had no effect on the expressions of caspase-9and MMP-9.The first two chapters determined the effect of RAAP in vitro, and furthermore, in order to confirm the effect of RAAP in vivo, the third chapter used xenografts of nude mice by pancreatic cancer BxPC-3cells, to investigate the effect of RAAP on the development of xenografts. The results showed that RAAP (10μg/kg) could inhibit the growth of xenografts significantly; by histological sections and TUNEL assay, the tumor cells in the RAAP group were apoptotic obviously; meanwhile, RAAP could promote the expressions of caspase-3and caspase-8, and reduce the expressions of VEGF-A and MMP-2, which were consistent with the results in vitro. Compared with the positive drug Gemcitabine, the efficacy of RAAP is similar, but the effective dose is significantly reduced.Taken together, RAAP could promote the apoptosis of pancreatic cancer cells by activating caspase-8and further caspase-3; RAAP could inhibit the angiogenesis by decreasing the expression of VEGF-A; RAAP could reduce the cell invasion by down-regulate the expression of MMP-2, which was the key gene for proteolysis of extracellular matrix; the results in vitro and in vivo were consistent. Currently, the clinical application of RAAP is still under phase II/III clinical trial, the investigation on RAAP anti-pancreatic cancer can supply the theoretic and laboratory evidences for the clinical application, also the new clues for the development of new drugs and the optimization of chemotherapy for pancreatic cancer. |
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