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目的:考察三种黄酮类化合物在乏氧条件下对卵巢癌SKOV3细胞的放射增敏效应。方法:将SKOV3细胞分为常氧组与乏氧组,建立体外SKOV3乏氧细胞模型并检测细胞内HIF-1α蛋白的水平加以验证;克隆形成法检测常氧与乏氧SKOV3细胞的放射敏感性;MTT法检测三种黄酮类化合物对SKOV3细胞增殖抑制的影响,筛选出药物的无毒浓度范围;改良的MTT法检测放射增敏效应,结合多靶单击模型拟合剂量存活曲线,分析放射生物学参数。结果:常氧组和乏氧组HIF-1α与β-Actin的灰度值之比分别为0.117和1.068,乏氧SKOV3细胞的放射敏感性显著低于常氧细胞;黄酮类化合物作用于SKOV3细胞的存活率随药物作用浓度的增大而降低;在常氧与乏氧条件下,随着药物浓度的增大,白杨素与槲皮素组D_0、D_q值显著减小(P<0.01),SER值显著增大(P<0.05)。灯盏花素各浓度组之间SER值无显著性差异(P>0.05)。乏氧组D_0、D_q、SER值整体高于常氧组。结论:乏氧诱导SKOV3细胞内HIF-1蛋白表达显著升高,导致细胞放射敏感性降低。白杨素与槲皮素可明显改善SKOV3细胞放射敏感性,且在乏氧条件下放射增敏效应更明显,而灯盏花素不具有此功效,其增敏机制可能是通过降低细胞内HIF-1α的水平和延迟细胞修复亚致死性损伤而实现。
OBJECTIVE: To investigate the radiosensitization effect of three flavonoids on ovarian cancer SKOV3 cells under hypoxic conditions. Methods: SKOV3 cells were divided into normoxia group and hypoxia group. SKOV3 hypoxia cells model was established and the level of HIF-1α protein in cells was verified. The radiosensitivity of normoxia and hypoxia SKOV3 cells was detected by clone formation assay MTT assay was used to detect the effect of three flavonoids on the SKOV3 cell proliferation inhibition. The non-toxic concentration range of the drug was screened out. The modified MTT method was used to detect the radiosensitization effect. The multi-target click model was used to fit the dose-response curve, Biological parameters. Results: The ratio of the gray values of HIF-1α and β-Actin in normoxia group and hypoxia group were 0.117 and 1.068, respectively. The radiosensitivity of hypoxia-exposed SKOV3 cells was significantly lower than that in normoxic cells. Flavonoids acted on SKOV3 cells (P <0.01). Under normoxia and hypoxia conditions, the D_0 and D_q values of chrysin and quercetin groups decreased significantly with the increase of drug concentration (P <0.01) SER value increased significantly (P <0.05). There was no significant difference in SER value between breviscapine groups (P> 0.05). The hypoxic group D_0, D_q, SER values were higher than the normoxic group. CONCLUSION: Hypoxia induces a significant increase in HIF-1 protein expression in SKOV3 cells, leading to a decrease in radiosensitivity. Chrysin and quercetin can significantly improve the radiosensitivity of SKOV3 cells, and the radiosensitization effect is more obvious under hypoxia, while breviscapine does not have this effect, and its sensitization mechanism may be through the reduction of intracellular HIF-1α Of the level and delay cell repair sublethal damage and achieve.