苯并[a]芘暴露的小鼠原代支气管上皮细胞中miR-320对细胞周期的影响

Effect of miR-320 on Cell Cycle of Primary Murine Bronchial Epithelial Cells Exposed to Benzo(a)pyrene

目的检测miR-320在苯并[a]芘(B[a]P)暴露的小鼠原代支气管上皮细胞的表达水平,探讨B[a]P染毒的小鼠支气管上皮细胞中miR-320对细胞周期的影响。方法以1.00μmol/LB[a]P染毒原代培养小鼠支气管上皮细胞。以定量PCR检测染毒细胞中miR-320的表达。以FACSArray流式液相芯片分析仪检测细胞周期和细胞周期素依赖性激酶6(CDK6)的表达水平。结果 miR-320在B[a]P染毒细胞中表达上调。染毒细胞出现G1期阻滞现象,与溶剂对照组的[(62.70±1.54)%]G1期细胞比例相比,(84.28±0.36)%的细胞停留在G1期。抑制miR-320表达后G1期阻滞现象缓解。染毒细胞中的CDK6表达降低,抑制miR-320表达后,与B[a]P组相比,CDK6的表达量有(2.0±0.3)倍的升高。结论 miR-320在一定程度上通过对CDK6表达的调控影响B[a]P暴露的细胞的细胞周期。

Objective To explore the role of miR-320 in cell cycle of primary murine bronchial epithelial ceils exposed to B[a]P by detecting the expression level of miR-320 in primary murine bronchial epithelial cells exposed to benzo（a）pyrene （B[a]P）. Methods The primary murine bronchial epithelial cells were cultured and exposed to 1.00 μmol/L B[a]P： The expression of miR-320 was detected by quantitative real time polymerase chain reaction assay. The cell cycle and the expression levels of cyclin-dependent kinases 6 （CDK6） were analyzed using FACSArray Flow cytometer. Results The expression of miR-320 was up-regulated in B[a]Pexposed cells. The B [a]P-exposed cells underwent G1 arrest, the percentage of cells in the G1 phase were （84.28±0.36）%, compared with that [（62.70±1.54）% ] of the control group. The relief of the G1 arrest was shown in anti-miR-320 group. The B[a]P-exposed cells resulted in reduced expression level of CDK6. The expressions of CDK6 in anti-miR-320 group was （2.0±0.3）-fold higher than that in B[a]P group. Conclusion MiR-320 may influence G1 arrest partially by regulating CDK6 in B[a]P-exposed cells.