Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag da...Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.展开更多
The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL)...The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in apoptosis of granulosa cells were explored. By using sodium prasterone sulfate rat PCOS model was induced. The apoptosis of granulosa cells in ovaries of rats was observed by TdT-mediated dUTP-biotin nick end-labeling (TUNEL), and the expression of TRAIL protein and mRNA in granulosa cells was detected by using immunhistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) respectively. The apoptotic rate and the expression of protein TRAIL in granulosa cells were significantly higher in antral follicles from the PCOS rats than in those from the control rats (P<0.01, P<0.05). There was no significant difference in apoptotic rate and the expression of TRAIL protein in granulosa cells of preantral follicles between the PCOS rats and the control rats (P>0.05). No apoptosis and the expression of TRAIL protein in granulosa cells of primordial follicles were found in the two groups. The expression of TRAIL mRNA was significantly stronger in granulosa cells from the PCOS rats than in those from the con- trol rats (P<0.01). It was suggested that the apoptotic rate in granulosa cells was significantly higher in antral follicle from the PCOS rats than in those from the control rats. TRAIL played a role in regu- lating the apoptosis of granulosa cells in PCOS rats.展开更多
文摘Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.
文摘The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in apoptosis of granulosa cells were explored. By using sodium prasterone sulfate rat PCOS model was induced. The apoptosis of granulosa cells in ovaries of rats was observed by TdT-mediated dUTP-biotin nick end-labeling (TUNEL), and the expression of TRAIL protein and mRNA in granulosa cells was detected by using immunhistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) respectively. The apoptotic rate and the expression of protein TRAIL in granulosa cells were significantly higher in antral follicles from the PCOS rats than in those from the control rats (P<0.01, P<0.05). There was no significant difference in apoptotic rate and the expression of TRAIL protein in granulosa cells of preantral follicles between the PCOS rats and the control rats (P>0.05). No apoptosis and the expression of TRAIL protein in granulosa cells of primordial follicles were found in the two groups. The expression of TRAIL mRNA was significantly stronger in granulosa cells from the PCOS rats than in those from the con- trol rats (P<0.01). It was suggested that the apoptotic rate in granulosa cells was significantly higher in antral follicle from the PCOS rats than in those from the control rats. TRAIL played a role in regu- lating the apoptosis of granulosa cells in PCOS rats.
