Preparation of Sheep (Ovis aries) and Cow (Bos taurus) Plancenta Immumoregulating Factor and Its Effects on Proliferation and Migration of PANC-Ⅰ Cells
Abstract：Placenta immunoregulating factor (PF) is composed of small polypeptides, which is extracted from the placenta. PF is valuable candidate for treatment of viral disease, immune deficiency disorder and malignant tumor. To detect the effects of sheep (Ovis aries) and cow (Bos taurus) placenta immunoregulating factor on the proliferation and migration of the human(Homo sapiens) pancreatic cancer cells-Ⅰ(PANC-Ⅰ) in vitro. PF was prepared from placenta by bienzyme hydrolysis. PANC-Ⅰ cells cultured in vitro were exposed to PF at the mass concentrations of 100~2 000 ng/mL. The effect of PF on the PANC-Ⅰ cell cycle was detected by flow cytometric method. The migration ability of PANC-Ⅰ cell was investigated with Transwell assay. The inhibition to proliferation rate of PANC-Ⅰ cells were evaluated by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay. The mRNA expression levels of tumor suppressor gene (P53) and cyclin-dependent kinase inhibitor 1A (P21) were analyzed by using real-time PCR. The relative expression was determined by using the comparative 2-??Ct method. The results showed that 0.588 mg PF derived from 17.11 g sheep placenta and 0.312 mg PF derived from 0.616g cow placenta, and the quality ratio was 20 000∶1. The morphology of the PANC-Ⅰ cells was not affect by adding the PF. The cell inhibition rate of PANC-Ⅰ cells was from 9.75%~22.89% with different concentration of sheep PF and from －3.81%~23.56% with different concentration of cow PF. The most remarkable concentration of sheep PF and cow PF were 500 ng/mL and 1 000 ng/mL, respectively. At the same time, the PI and SPF of PANC-Ⅰcells was the lowest with 500 ng/mL of sheep PF, but the proliferation index (PI) and S-phase fraction (SPF) were not affected by cow PF. The migration ability of PANC-Ⅰcells was not affected by PF. The expression levels of P53 and P21 of PANC-Ⅰcells were increased and decreased respectively after the treatment of PF. The results showed that the PF had a negative effect on the proliferation of PANC-Ⅰ cells in vitro, and the different effects of sheep and cow PF on PANC-Ⅰcells may related to the composition of the PF. This study provides new ideas for the treatment of pancreatic cancer.
