Thrombospondins (TSP-1, TSP-2) are secretory extracellular glycoproteins that are involved in a variety of physiological processes such as tumor cell adhesion, invasion, and metastasis. The present study was undertaken to elucidate the involvement of thrombospondins in the adhesion of osteoblast-like cells using the TSP-1 or TSP-2 antisense MG63 and MC3T3-E1 cell lines. For downregulation of TSPs expression, we prepared antisense constructs for TSP-1 and TSP-2 using the pREP4 an episomal mammalian expression vector, which be able to produce the specific antisense oligonucleotides around chromosome. MG63 and MC3T3-E1 osteoblast-like cells were transfected with the antisense constructs and nonliposomal Fugene 6, and then selected under hygromycin B (50 μM/mℓ) treatment for 2 weeks. Western blot analysis revealed that expression of the TSP proteins was downregulated in the antisense cell lines. The cell adhesion assay showed that adhesive properties of TSP-1 and TSP-2 antisense MG63 cells on the polystyrene culture plate were reduced to 17% and 21% of the control cells, respectively, and those of the TSP-1 and TSP-2 antisense MC3T3-E1 cells also decreased to 19% and 27% of control, respectively. Adhesion of TSP-1 and TSP-2 antisense MC3T3-E1 cells on Type I collagen-coated culture plate decreased to 27% and 76%, respectively. These results indicate that TSP-1 and TSP-2 proteins may have an important role in adhesion of osteoblast-like cells to extracellular matrix.

The different forms of flowers in a species have drawn thoughtful attention of many evolutionary botanists, including Charles Darwin. Common buckwheat (Fagopyrum esculentum Moench.) is regarded as a dimorphic self-incompatible plant which bears either a pin or a thrum flower. It is revealed that the S supergene the key element to govern the self-incompatibility, flower morphology, and pollen size. Already, we have produced self-incompatible buckwheat lines by an interspecific cross between F. esculentum and F. homotropicum by using embryo rescue. We also notice that the self-compatibility allele, Sh, keeps up the heteromorphic incompatibility. In the past decades, two dimensional gel electrophoresis based proteomics approaches have been applied systematically to identify and profile proteins expressed during pollen development of model plant species. Proteome techniques have vastly been applied in the fields of plant genetics, plant development, and plant physiology and ecology to reveal plant genetic diversity, plant development, differentiation of plant tissue and organ, separation and functional identification of novel component of various organelles, mechanisms of plant adapted to abiotic or biotic stresses including high temperature, low temperature, high salt, drought, and pathogens and insects, and interaction of plant with microbe. However, the plethora of studies related to heteromorphic has added remarkably to our knowledge in the field of the multiple aspects of the breeding system and many researches have provided evidence for the connection between these two components. But in spite of its potential biological significance, the high throughput proteomics analysis of this connection has so far been grossly overlooked. So our attempts are to unravel the proteome investigation in common buckwheat.