ABSTRACT
ObjectiveTo observe the effect of Danshen injection （DAN） on platelet （PLT）-induced metastasis of breast cancer cells in vitro. MethodThe 3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide （MTT） assay was used to observe the effect of DAN on the growth of MDA-MB-231 cells in vitro. Oris™ migration assay was used to determine the effect of DAN （final mass concentrations 4， 8， 16 g·L-1） on PLT （1.5×1010 cells/L）-induced migration of breast cancer cells in vitro. The effect of DAN on PLT-induced cell invasion was detected by Transwell assay. Immunofluorescence and Western blot were used to detect the effect of DAN on the protein expression associated with PLT-induced epithelial-mesenchymal transition （EMT）. In addition， enzyme-linked immune-sorbent assay （ELISA） was used to determine the effect of DAN （final mass concentrations 4， 8， 16， 32， 64 g·L-1） on the secretion of transforming growth factor-β1 （TGF-β1）. Western blot was used to observe the effect of DAN on the expression of podoplanin （PDPN） protein in MDA-MB-231 cells induced by PLT. ResultCompared with the blank group， the DAN groups （32 and 64 g·L-1） showed decreased A570 （P<0.05， P<0.01）， and there was no significant difference in A570 between DAN groups （4， 8， 16 g·L-1）. Compared with the blank group， the PLT group showed increased cell migration and invasion， while DAN groups significantly inhibited PLT-induced cell migration and invasion. Compared with the blank group， the PLT group showed decreased expression of E-cadherin， while DAN could significantly reverse this effect of PLT. Compared with the blank group， the PLT group showed increased Slug and Snail protein expression （P<0.05， P<0.01）， while DAN significantly reversed Snail protein expression induced by PLT （P<0.05， P<0.01）. The content of TGF-β1 in the PLT group increased （P<0.01）， while the secretion of TGF-β1 induced by PLT decreased in the DAN groups （16， 32， and 64 g·L-1） （P<0.05， P<0.01）， and the secretion of TGF-β1 was not significantly affected in other DAN groups. PDPN protein expression in the PLT group increased （P<0.01）， while DAN could significantly inhibit PLT-induced PDPN expression （P<0.01）. ConclusionDAN can inhibit PLT-induced migration， invasion， and EMT of breast cancer cells. The mechanism may be related to the direct action between breast cancer cells and tumor cells by down-regulating PDPN expression and interfering with PLT and has nothing to do with the effect of TGF-β1 secretion of PLT.
ABSTRACT
Thromboxane A2 receptors (TPs) widely distribute in different organ systems and localize both on cell membranes and in intracellular structures.TPs are the members of seven-transmembrane G-protein-coupled receptor (GPCR) super family.Historical-ly, the involvement of TPs in platelet functions has received the greatest attention .TPs have the capacity to activate different signaling cascades which regulate platelet shape change , aggregation and secretion response .Currently , anti-platelet drugs primarily act on re-ceptors and /or signaling molecules in activation pathways .The signaling transduction of TPs in platelet contributes to the investigation of the effects of extracts of traditional Chinese medicine on antiplatelet aggregation and the exploration of the action mechanisms .
ABSTRACT
Editor's note: The research on effective constituents of traditional Chinese medicine is one of the crucial areas of basic research of traditional Chinese medicine. It is also a research hotspot which gains widespread attention and active application of projects of national natural science funds in the discipline of traditional Chinese medicine. In 2013, the National Natural Science Foundation of Chinese traditional medicine in the application code H2803 in the research field of effective constituents of traditional Chinese medicine , the direction of gener-al project , Youth Science Foundation and the regional Science Foundation project application 449 , which accept-ed 425 . In this paper , the general situation of the application and approved projects of national natural science funds in the research field of effective constituents of traditional Chinese medicine in 2013 has been introduced. The research strategies and plans of the approved projects have been summarized , and the problems of the ap-plications have been also analyzed .