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.
ABSTRACT
This study examined the effect of saponins from Tupistra chinensis Bak (STCB) on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms.Cell proliferation was assessed by MTT assay.Cell cycle distribution was determined by flow cytometry.Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 μg/mL 5-fluorouracil (5-Fu) as a positive control.The activity of nuclear factor (NF)-κB was detected by gel mobility shift assay.The mRNA level of NF-κB was determined by real-time quantitative RT-PCR.The results showed that in vitro STCB inhibited the growth of S-18 0 cells in a concentration-dependent manner,which was accompanied by cell cycle arrest at S-phase.In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis.Moreover,STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts.It was concluded that STCB inhibits the proliferation and cell cycle progression of S-180 cells by suppressing NF-κB signaling in mouse xenografts.Our findings suggest STCB is a promising agent for the treatment of sarcoma.
ABSTRACT
This study examined the effect of saponins from Tupistra chinensis Bak (STCB) on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms.Cell proliferation was assessed by MTT assay.Cell cycle distribution was determined by flow cytometry.Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 μg/mL 5-fluorouracil (5-Fu) as a positive control.The activity of nuclear factor (NF)-κB was detected by gel mobility shift assay.The mRNA level of NF-κB was determined by real-time quantitative RT-PCR.The results showed that in vitro STCB inhibited the growth of S-18 0 cells in a concentration-dependent manner,which was accompanied by cell cycle arrest at S-phase.In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis.Moreover,STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts.It was concluded that STCB inhibits the proliferation and cell cycle progression of S-180 cells by suppressing NF-κB signaling in mouse xenografts.Our findings suggest STCB is a promising agent for the treatment of sarcoma.
ABSTRACT
Background The main mechanism of diabetic retinopathy (DR) is the formation of retinal neovascularization. Vascular endothelial growth factors (VEGF) is primarily factor for DR. Objective The goal of this study was to investigate the effects of angelicae sinensis decoction for supplementing blood ( ASD), a kind of traditional prescription of Chinese herbal medicine, on vascular endothelial growth factor (VEGF) level in vitreous and serum of STZ-induced type 2 diabetes mellitus. Methods Thirty-four SPF male SD rats were fed using high fat diet for 8 weeks and then the streptozocin of 25 mg/kg was intraperitoneally injected to create the type 2 diabetic models. ASD (3.6 g/kg) and 10 mg/kg Gli of 5 mi were administered intragastrically from the 4th day of high fat die though 8 weeks respectively. Other 10 matched rats were used as normal control group. The body weight, blood glucose level ,triglycerides (TG) and high density lipoprotein (HDL) were detected in 4 and 8 weeks after usage of drug. The vitreous and serum samples were obtained in the 8 th weeks for the VEGF detection by enzyme-linked immunosorbent assay (ELISA). This experiment comlied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results The body weight and HDL in DM group were significantly lower than those of normal control group in 4 or 8 weeks ( P＜0. 01 ), but in ASD and Gli groups ,the body weight and HDL were higher than those of the DM group (P＜0. 05 ). the blood glucose level and TG were elevated in DM group compared with normal control group (P＜0. 01 ), but those in ASD group and Gli group were decreased in comparison with DM group ( P＜0.05 ). EIISA revealed that no significant differences were found in VEGF levels between normal control group and ASD group or Gli group (P＞0. 05 ), but in DM group, the serum VEGF level (147.65±4. 55 ng/L) was reduced and vitreous VEGF level (673.45± 100. 85 ng/L) was raised,showing the considerable differences (P＜ 0.05 ). No any correlation was found between serum VEGF level and vitreous VEGF level (r=0.314,P＞0. 05). Conclusion ASD can decrease the VEGF level in vitreous,implying that ASD postpone the occurrence of diabetic retinopathy in DM rat.