Effect of Wumeiwan-medicated Serum on Proliferation， Invasion， Migration， Apoptosis of Pancreatic Cancer Cells Based on PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo observe the effects of Wumeiwan-medicated serum on the proliferation， invasion， migration， and apoptosis of human pancreatic cancer SW1990 cells and explore the underlying mechanism. MethodThe Wumeiwan-medicated serum was prepared and the pancreatic cancer SW1990 cell line was cultured in vitro. The optimal time of Wumeiwan-medicated serum was selected for subsequent experiments by cell counting kit-8（CCK-8）. SW1990 cells were divided into a control group and low- （2%）， medium- （4%）， and high-dose （8%） Wumeiwan-medicated serum groups. The colony-forming， migration， and invasion abilities were detected by clonogenic assay， wound healing assay， and transwell migration assay. Flow cytometry was used to detect the effect of Wumeiwan-medicated serum on the apoptosis of pancreatic cancer SW1900 cells. Western blot was used to detect the expression levels of apoptosis-related proteins， such as B-cell lymphoma-2（Bcl-2）， Bcl-2-associated X protein （Bax）， cytochrome C （Cyt C）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）， cleaved cysteinyl aspartate-specific protease-9 （cleaved Caspase-9）， as well as phosphatidylinositol 3-kinase（PI3K）， phosphorylated PI3K（p-PI3K）， protein kinase B （Akt）， and phosphorylated Akt（p-Akt）in PI3K/Akt pathway in SW1990 cells. ResultCompared with blank group， Wumeiwan groups showed decreased absorbance （A） 72 h after drug intervention （P<0.01）. Compared with the low-dose group， the medium- and high-dose groups showed decreased A （P<0.01）. Compared with the medium-dose group， the high-dose group showed decreased A （P<0.01）. It indicates that Wumeiwan can inhibit SW1990 cell proliferation in a dose-dependent manner after 72 h， and the optimal action time is 72 h. Compared with the blank group， the Wumeiwan groups showed weakened invasion of SW1990 cells （P<0.01）， reduced colony-forming and migration abilities （P<0.05， P<0.01） in a dose-dependent manner， and increased total apoptosis rates （P<0.01）. The inducing effect of Wumeiwan on apoptosis increased with the increase in dosage. Compared with the blank group， the Wumeiwan groups showed decreased protein expression of Bcl-2 （P<0.01）， increased protein expression of cleaved Caspase-3， cleaved Caspase-9， Cyt C， and Bax （P<0.05， P<0.01） in a certain dose-effect relationship， reduced protein expression of p-PI3K and p-Akt （P<0.05， P<0.01） with the increase in dosage， and declining p-PI3K/PI3K and p-Akt/Akt （P<0.05， P<0.01） with the increase in dosage. ConclusionWumeiwan-medicated serum can significantly inhibit the malignant biological behaviors of pancreatic cancer SW1990 cells and induce apoptosis. The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway and down-regulation of protein phosphorylation level in the PI3K/Akt signaling pathway.