This study investigated the molecular markers of DS-1-47,a component of an implantation-promoting traditional Chinese medicine consisting of Astragalus mongholicus,Atractylodes macrocephala,Scutellaria baicalensis and Dipsacales,in an attempt to clarify the molecular mechanism and action targets of DS-1-47.Controlled ovarian stimulation(COS) method was used to establish the implantation dysfunction models of mice.Animals were divided into normal pregnant group,COS model group and DS-1-47 group.Laser capture microdissection-double dimensional electrophoresis-mass spectrum(LCM-DE-MS) was used to analyze the uterine protein molecules that were possibly involved in the promotion of implantation.Twenty-three proteins in DS-1-47 group were significantly changed as compared to those in COS model group,with 7 proteins down-regulated and 16 proteins up-regulated.Except for some constituent proteins,the down-regulated proteins included collagen α-1(Ⅵ) chain,keratin 7,keratin 14,myosin regulatory light chain 12 B,myosin light polypeptide 9,heat shock protein β-7,and C-U-editing enzyme APOBEC-2;the up-regulated proteins included apolipoprotein A-I,calcium regulated protein-3,proliferating cell nuclear antigen,L-xylulose reductase,and calcium binding protein.These 23 proteins that were regulated by DS-1-47 represented a broad diversity of molecule functions.The down-regulated proteins were associated with stress and immune response,and those up-regulated proteins were related to proliferation.It was suggested that these proteins were important in regulating the uterine environment for the blastocyst implantation.By identification of DS-1-47 markers,proteomic analysis coupled with functional assays is demonstrated to be a promising approach to better understand the molecular mechanism of traditional Chinese medicine.
Neamine, a non-toxic derivative of neomycin, has recently been shown to have antitumor activities in various types of cancers. However, its effect on pancreatic cancer is still unknown. The study aimed to investigate its antitumor activity on pancreatic cancer and the underlying mechanisms. MTT assay was used to observe the effect of neamine on angiogenin(ANG)-induced As PC-1 cell proliferation. Tissue microassay and immunofluorescence staining were used to detect the expression of ANG and its nuclear translocation, respectively. Tumor xenografts were established by subcutaneous inoculation of As PC-1 pancreatic cancer cells into the right flanks of nude mice, and neamine was injected subcutaneously. Immunohistochemistry was done to observe the expression of ANG, CD31 and Ki-67 in tumor xenografts. It was found that neamine blocked the nuclear translocation of ANG effectively and inhibited ANG-induced As PC-1 cell proliferation in a dose-dependent manner. Neamine had anti-tumor effects on As PC-1 xenograft models. Consistently, neamine reduced the expression levels of ANG, Ki-67 and CD31 in tumor xenografts. It was concluded that neamine may be a promising agent for treatment of pancreatic cancer.