骨连接素对角质形成细胞的增殖抑制作用

Anti-proliferative effects of osteonectin on keratinocytes

目的:观察骨连接素对体外培养人角质形成细胞(HaCaT)的细胞增殖、细胞周期及相关基因表达的影响,初步探讨骨连接素对人角质形成细胞的作用机制。方法:用不同浓度骨连接素处理HaCaT细胞,MTT法检测HaCaT细胞增殖;流式细胞术测定细胞周期;实时定量RTPCR方法检测CCND1和PCNAmRNA表达水平的变化。结果:与未加入骨连接素的HaCaT细胞相比,5μg/ml以上浓度骨连接素作用的细胞增殖率明显下降(P<0.05),10μg/ml骨连接素作用的细胞G0/G1期细胞明显增多。10μg/ml以上浓度的骨连接素作用后,HaCaT细胞CCND1和PCNA的mRNA水平显著减弱(P<0.05,P<0.01)。结论:骨连接素可能是通过抑制CCND1和PCNA的表达,从而发挥抑制细胞增殖的作用。

Objective： To investigate the effects of osteonectin on cell proliferation, ceil cycle and related gene expression of in vitro cultured HaCaT cell line for the purpose of studying the action mechanism of osteonectin on keratinocytes. Methods：HaCaT cells were treated with serial concentrations of osteonectin. Cell proliferation was assessed by MTT and cell cycle was analyzed by flow cytometry. The mRNA levels of cell cycle regulatory gene CCND1 and PCNA were detected by real-time RTPCR. Results： Compared with control group （without using osteonectin） ,osteonectin at〉5 μg/ml effectively inhibited the proliferation of HaCaT cells（P〈0.05） and caused G0/G1 phase arrest at 10μg/ml （P〈0.05）. The mRNA levels of CCND1 and PCNA were significantly decreased after treatment with osteonectin at〉10μg/ml（P〈0.05, P〈0.01）. Conclusion： Osteonectin can effectively inhibit cell cycle progression and proliferation of HaCaT cells by decreasing the expression of CCND1 and PCNA gene.