Journal of International Reproductive Health/Family Planning ›› 2016, Vol. 35 ›› Issue (2): 96-100.

• 论著 • Previous Articles     Next Articles

Construction of LV-calb2 and LV-siRNA-calb2 and Expression in Leydig Cells of Teste

XU Wen-dan,DAI Xiao-nan,ZHU Qian,ZHANG Bei,GAO Chao,GAO Li,LIU Jia-yin,CUI Yu-gui   

  1. Clinical Center of Reproductive Medicine,First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China
  • Received:1900-01-01 Revised:1900-01-01 Published:2016-03-15 Online:2016-03-15
  • Contact: CUI Yu-gui

Abstract: Objective:To explore the effect of CALB2 on steriodogenesis in testicular Leydig cell,we firstly established LV-calb2 and LV-siRNA-calb2 lentivirus vectors. Methods:Firstly,the over-expression and interference plasmids were synthysized,and the bacterial competent cells were prepared ahead. Secondly,polymerase chain reaction(PCR) and gene sequencing were administered to ensure the LV-calb2 and LV-siRNA-calb2 lentivirus vectors constructed successfully. At last,the infection efficiency was tested and then the level of testosterone in MLTC-1 and the level of progesterone in R2C were measured by radioimmunaossay after MLTC-1 and R2C were infected with LV-calb2 and LV-siRNA-calb2 respectively. Results:PCR and gene sequencing confirmed that LV-calb2 and LV-siRNA-calb2 lentivirus vectors were established,with the capacity to infect efficiently MLTC-1 and R2C. Testosterone production was significantly increased in the MLTC-1 infected with LV-calb2 (P<0.001), while progesterone production was significantly decreased in R2C infected with LV-siRNA-calb2(P<0.05). Conclusions:This study successfully constructed LV-calb2 and LV-siRNA-calb2 lentivirus vectors which could infect Leydig cells with high-efficiency. The preliminary result indicated that CALB2 promoted steriodogenesis in Leydig cells.

Key words: Calcium-binding protein, vitamin D-dependent, Lentivirus, Testis, Leydig cells, Androgens