角质形成细胞生长因子及其受体反义寡核苷酸对HaCaT细胞的影响

The Effects of Keratinocyte Growth Factor and Its Receptors Antisense Oligonucleotide on Prolifera- tion of HaCat Cells

目的研究角质形成细胞生长因子(KGF)及其受体(KGFR)反义寡核苷酸对KGF介导的细胞周期和凋亡变化的抑制作用。方法利用永生化角质形成细胞株-HaCaT细胞进行体外培养,流式细胞仪检测KGF引起的HaCaT细胞细胞周期和凋亡变化以及KGFR mRNA反义寡核苷酸对KGF的抑制作用。结果 10ng/mL KGF刺激24 h后可使S期比率较自然生长组增加15%(P＜0．05),并伴凋亡增加2．9%(P＜0．01)。针对KGFR mRNA不同位点的2个序列反义寡核苷酸作用后,S期比率和凋亡率降低,且抑制作用呈浓度依赖性(P均＜0．05)。反义寡核苷酸作用后G_0G_1期相应增加,也呈浓度依赖性(P＜0．01)。各实验组G_2M期比率差异无统计学意义(P均＞0．05)。结论 KGF可能是造成银屑病KC增殖和凋亡的诱因。KGFR mRNA反义寡核苷酸对KGF介导的HaCaT细胞增殖、凋亡增加具有抑制作用。

Objective To investigate the effects of keratinocyte growth factor （KGF） and KGF receptor （KGFR） antisense oligonucleotide （ASODN） on cell cycle and apoptosis of HaCat cells. Methods HaCaT cell, an immortalized keratinocyte cell strain, was cultured in vitro. Flow cytometry was used to measure the cell cycle and apoptosis mediated by KGF and ASODN. Results The rates of S phase and apoptosis in the group treated with KGF increased significantly than those in the control group （both P 〈 0.01 ）. With the treatment of KGFR mRNA ASODN the increase of the rates of S phase and apoptosis induced by KGF were significantly suppressed （P 〈 0.01, P 〈 0.05 respectively） in a close-dependent manner （r=0.99, P 〈 0.05 ）. Conclusions KGF may play an important role in cell cycle and apoptosis of KC, while KGFR mRNA ASODN is an effective inhibitor of proliferation and apoptosis mediated by KGF, which may make a sense in the treatment of psoriasis.