FOXO4-DRI多肽增加非小细胞肺癌细胞放射敏感性研究

FOXO4 D-retro-inverso peptide increases radiosensitivity of non-small cell lung cancer cells

目的探讨FOXO4-DRI多肽对非小细胞肺癌(NSCLC)细胞放射敏感性的影响。方法为检测FOXO4-DRI对NSCLC细胞的影响,将H460和A549细胞按照射与给药方式分为对照组、FOXO4-DRI组、单纯照射组、照射+FOXO4-DRI组。采用剂量率为0.99 Gy/minγ射线单次照射,在照射前10 min对H460细胞给予FOXO4-DRI 6μmol/L,A549细胞给予FOXO4-DRI 30μmol/L。采用CCK-8法检测照射后24、48和72 h细胞存活率;用结晶紫染色法检测细胞克隆形成数目;用伤口愈合实验检测细胞迁移程度;用流式细胞术检测细胞凋亡和细胞周期的改变。结果与对照组相比,FOXO4-DRI组H460细胞和A549细胞存活率降低(t=1.06~50.75,P<0.05)、迁移率降低(t=33.37~139.10,P<0.05),克隆形成数目减少(t=5.20~93.48,P<0.05)。FOXO4-DRI诱导了细胞凋亡(t=2.95~42.00,P<0.05),导致G0/G1细胞周期阻滞以及G2/M期细胞比例下降(t=3.50~31.59,P<0.05)。与照射组相比,FOXO4-DRI可进一步降低辐照后的细胞存活率(t=2.94~23.40,P<0.05),减少辐照后克隆形成数目(t=8.43~34.00,P<0.05)和细胞迁移率(t=5.25、7.56,P<0.05),增加细胞凋亡(t=9.20~11.52,P<0.05)。照射+FOXO4-DRI组细胞,G2/M期细胞比例进一步下降,S期细胞增加(t=3.85~17.62,P<0.05)。结论FOXO4-DRI多肽通过促进细胞凋亡,降低细胞增殖能力和迁移率,可以提高NSCLC细胞放射敏感性。

Objective To explore the effects of FOXO4 D-retro-inverso peptide(FOXO4-DRI)on the radiosensitivity of non-small cell lung cancer(NSCLC)cells.Methods To detect the effect of FOXO4-DRI on NSCLC cells,H460 and A549 human lung cancer cells were divided into four groups,including untreated control,FOXO4-DRI,γ-ray irradiation and FOXO4-DRI+γ-ray groups.A sigle dose rate of 0.99 Gyγ-rays was used for radiation.H460 cells were administered with 6μmol/L FOXO4-DRI and A549 cells were adiminstered with 30μmol/L FOXO4-DRI at 10 min before radiation.Cell viability and survival were detected by CCK-8 assay and colony formation assay,respectively.Cell migration was detected by wound healing assay.Apoptosis and cell cycle arrest were detected with flow cytometry.Results FOXO4-DRI inhibited growth of H460 and A549 cells(t=1.06-50.75,P<0.05),and decreased cell mobility(t=33.37-139.10,P<0.05)and colony formation(t=5.20-93.48,P<0.05).FOXO4-DRI also increased apoptosis(t=2.95-42.00,P<0.05)and caused a cell cycle arrest at G0/G1 phase accompanied with a decreased proportion of G2/M phase(t=3.50-31.59,P<0.05).Furthermore,FOXO4-DRI increased radiosensitivity of both H460 cells and A549 cells(t=2.94-23.40,P<0.05),caused a Further Decrease of radiation-mediated mobility(t=5.25,7.56,P<0.05)and colony formation(t=8.43-34.00,P<0.05)and a more increase of radiation-induced apoptosis(t=9.20-11.52,P<0.05).FOXO4-DRI also further decreased the proportion of G2/M phase cells but increased the proportion of S phase cells(t=3.85-17.62,P<0.05).Conclusion FOXO4-DRI increases radiosensitivity of NSCLC cells by inducing apoptosis and suppressing cell proliferation.