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

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.