The study of the expressions of apoptosis and its related protein by knockdown of human immunodeficiency virus-1 vpr gene / 中华传染病杂志

ABSTRACT

Objective To observe the expression of apoptosis related protein c-inhibitor of apoptosis protein (IAP)2 by RNA interference of human immunodeficiency virus-1 (HIV-1)vpr gene and analyze the apoptosis of Jurkat cells.Methods Vector (NC),HIV-1vpr (vpr),vpr+ pRNAT-U6.1/Neo-vpr-56 (Si56) and vpr+ pRNAT-U6.1/Neo-vpr-160 (Si160) were transfected to Jurkat cells and cultured for 48 hours.The total RNA and protein were extracted.Expression of vpr gene was detected by reverse transcription (RT)-polymerase chain reaction (PCR) to confirm the success of transfection.Expression of c IAP2 was detected by RT-PCR and Western Blot.The apoptosis of Jurkat cells was observed by flow cytometry.Results Expression of vpr gene was detected in vpr,Si56 and Si160 groups.The mRNA expression levels in Si56 and Si160 groups were significantly lower than that in vpr group,which declined 87.2％ and 82.2％,respectively (P＜0.05).The levels of c IAP2 mRNA expression in vpr,Si56 and Si160 groups were increased by 3.75,2.49 and 2.65 folds,respectively,compared to that in NC group.However,the c-IAP2 mRNA expressions in Si56 and Si160 groups were lower than that in vpr group,which declined 33.7％ and 29.5％,respectively (P ＜ 0.05).The c-IAP2 protein expression was consistent with mRNA by immunoblotting,and those in Si56 and Si160 groups were declined 42.2％ and 46.8％,respectively,compared to that in vpr group (P＜0.05).The apoptosis of J urkat cells was detected in all groups.Compared to NC group,the apoptotic rate in vpr group was increased by 1.76 folds.However,the differences of apoptosis rate among NC,Si56 and Si160 groups were not statistically significant (all P＞0.05).Compared to vpr group,the apoptotic rates in Si56 and Si160 were significantly decreased by 19.26％ and 18.05％,respectively (P＜0.05).Conclusions The expression of c-IAP2 could be downregulated by knockdown of HIV-1 vpr gene in transcription and protein levels,and the apoptosis of Jurkat cells is inhibited.