Eukaryotic Expression Of ActivinA And The ActivinA-relative Antitumor Mechanisms For Traditional Chinese Medicines Compounds

Activin was isolated from the follicular fluid of the pig in 1986, which is constitutived by dimeride glucoprotein, which is composed from disulfide linkage and two pieces ofβsubunits. Activin has the typical structural feature of the transforming growth factor-βsuperfamily, so activin belongs to TGF-βsuperfamily members. According to the difference of conjunct formal subunit, which emerges activinA,activinB,activinAB. they have semblable biologic activity, Activin are a kind of multifunctional factors, they have tissue specificity, and have critical regulatory contribution in the tissue formation,cell growth,cell differentiation,development and maturation. Which also are the important regulatory factor in organism during physiology and pathology. The eukaryotic expression of activinA has important significances for investigating activin signal transduction passway,physiology and pathology procedure, it also established a essential basis for activin relative cancer investigation and treatment. In the present study, the recombinant plasmid pcDNA3-ActivinβA was transfected to CHO cells, and obtained cell strain of stable expressing activinA under the selection pressure of G418 in three weeks. then, we further confirmed the cell strain of overexpression activinA through report assay and western blot method。Because activin can inhibit proliferation of epithelial cells, it is proposed as a tumor suppress gene. Therefore, interfering of the activin signal transduction may result in tumorigenesis. Our previous study revealed that testes specific protein 50 (TSP50) promote tumoerigenesis via inhibiting acitivin signal transduction, our result also revealed that chinese medicine compound TI13, TI33, TI176 and TI179 can inhibit TSP50 expression and have significant anti-tumor activities, it tempt us to investigate the effect of these compounds on activin signal transduction. We transfected activin response reporter vector CAGA-lux into 293T cell, and added these traditional Chinese medicine compound to the 293T culture medium respectively. The result shown that compound TI13, TI176 and TI179 can enhance activin signal transduction. This paper established a experimental basis for investigation of activin function and mechanism in tumorigenesis.