BmNPV对家蚕BmN细胞周期及周期蛋白基因表达水平的影响

Effect of BmNPV on BmN cell cycle and expression level of Cyclinin silkworm

【目的】研究家蚕核型多角体病毒(Bombyx mori Nucleopolyhedroviruses,BmNPV)对家蚕卵巢细胞系(BmN)细胞周期及细胞周期蛋白基因转录水平的影响。【方法】用BmNPV感染家蚕BmN细胞,分别在病毒感染后的不同时间(12,24,36,48h),应用实时荧光定量PCR方法检测细胞周期蛋白基因CyclinA、CyclinB、CyclinE的表达情况,用流式细胞仪检测细胞周期的变化。【结果】BmNPV感染24h后,BmN细胞出现明显感染症状,变为圆形;48h后细胞核内出现多角体。随着感染时间的延长,CyclinA、CyclinB、CyclinE基因表达量均下降,分别在感染12,24,36h后,表达量降低为0。流式细胞仪检测结果显示,在BmNPV感染后24h左右,抑制于G2/M期的BmN细胞达到68%。【结论】BmNPV感染家蚕BmN细胞后,导致CyclinB、CyclinE基因表达量降低,对CyclinA基因表达量的影响较小;同时BmNPV感染可以将BmN细胞发育抑制于G2/M期。

[Objective] This study aimed to study the effect of BmNPV （Bombyx mori Nucleopolyhe- droviruses） on the cell cycle and expression level of Cyclin in silkworm BmN cell line. [Method] BmN cell was infected with BmNPV, and the transcription levels of CyclinA,CyclinB and CyclinE genes were detec- ted by QRT-PCR at different times （12,24,36, and 48 h）. Flow cytometer was used to detect the change of cell cycle. [Result] BmN cells showed obvious infection symptoms 24 h after infection and the shape of cells changed to circle. Polyhedrosis appeared in the nucleus 48 h after infection. As the increase of time af- ter viral infection,the expression levels of CyclinA,CyclinB and CyclinE decreased. The expression levels of CyclinA, CyclinB, CyclinE reduced to zero at 12,24 and 36 h, respectively. Flow cytometer results showed that 68% cells were inhibited at the G2/M phase 24 h after infection. [Conclusions] Infection of BmNPV reduced the expression levels of Cyclin. The effect on CyclinA was less than that on CyclinB and CyclinE. Infection of BmNPV inhibited BmN cells at G2/M phase.