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目的探讨HIV Tat-蛋白是否可以在细胞内影响主要节律基因Clock,Cry,Bmal1,Per的表达。方法用lipofectamineTM2000将HIV Tat表达质粒转入PC-12细胞,使Tat在PC-12细胞中表达。转染48h后提取mRNA和蛋白,分别通过实时荧光定量PCR技术和Western-blot技术检测节律基因在mRNA和蛋白水平表达的变化。结果研究结果显示,与未处理组和空质粒组相比,转染组Cry1的表达水平显著升高(P<0.05);Clock的表达水平显著降低(P<0.05);Bmal1和Per的表达水平无明显变化(P>0.05)。结论 HIV Tat影响了近日节律基因Cry1和Clock的表达,表现为Cry1表达水平升高,Clock表达水平下降,提示TAT蛋白可以影响节律基因的表达并且可能通过对近日节律的影响调节细胞生理功能。
Objective To investigate whether HIV Tat-protein can affect the expression of major rhythm genes Clock, Cry, Bmal1 and Per in the cell. Methods HIV Tat expression plasmids were transfected into PC-12 cells by lipofectamineTM2000 to express Tat in PC-12 cells. MRNA and protein were extracted 48h after transfection, and the changes of rhythmic gene mRNA and protein expression were detected by real-time fluorescence quantitative PCR and Western-blot respectively. Results The results showed that compared with untreated group and blank plasmid group, the expression of Cry1 in transfected group was significantly increased (P <0.05); the expression of Clock was significantly decreased (P <0.05); the expression of Bmal1 and Per No significant change (P> 0.05). Conclusion HIV Tat affects the expression of Cry1 and Clock in recent rhythmic genes. Cry1 expression and Clock expression decrease. It suggests that TAT protein may affect the expression of rhythmic genes and may regulate the cell physiological function through the influence of rhythms.