茉莉素对人神经母细胞瘤细胞株SK—N—SH的抑制增殖及诱导凋亡作用

Jasmonates suppress proliferation and induce apoptosis in human neuroblastoma cell line SK-N-SH

目的观察茉莉素对人神经母细胞瘤细胞株SK—N—SH的抑制增殖及诱导凋亡作用，并比较茉莉酸甲酯、顺式茉莉酮、茉莉酸作用强度差异。方法1～10mmol／L茉莉酸甲酯、顺式茉莉酮、茉莉酸分别处理SK-N-SH细胞后，NTT法检测细胞生长活性；克隆形成实验检测细胞增殖能力；PI单染法流式细胞术检测细胞周期；AO／EB荧光染色法、Hoechst33258荧光染色法、Annexin V-FITC／PI双染法流式细胞术检测细胞凋亡。结果各浓度茉莉素作用后，SK-N—SH细胞呈时间、浓度依赖性生长抑制，其中茉莉酸甲酯作用最强，1～10mmol／L茉莉酸甲酯作用24h后，细胞生长抑制率为（22．74～88．67）％，IC50为1．44mmol／L；2．5mmol／L茉莉酸甲酯、顺式茉莉酮、茉莉酸作用后，细胞克隆形成能力分别降低87．00％、78．66％、39．00％，细胞周期示S期细胞比率显著增高，G0／G1期细胞比率下降，部分细胞发生典型的凋亡形态学改变；1．5～2．5mmol／L茉莉酸甲酯作用后，细胞凋亡率为（23．77—36，43）％。结论茉莉素能通过减弱瘤细胞增殖、诱导细胞周期阻滞和凋亡等机制抑制人神经母细恩情瘤细胞株的生长活性。

Objective To explore the growth inhibition and apoptosis induction by jasmonates on human neuroblastoma SK-N-SH cell line, and to compare the anti-tumor effects of methyl jasmonate, cisjasmone and jasmonic acid. Methods After administration of 1-10 mmol/L jasmonates for 6-24 h, the growth inhibition rate of SK-N-SH cells were studied by MTF colorimetry. Cell proliferation ability was detected by colony formation assay. Cell cycle phases were checked by propidium iodide staining flow cytometery （FCM）. Cell apoptosl, was detected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and Annexin V-FITC and propidium iodide staining flow cytometry. Resuits Jasmonates could inhibit the growth of SK-N-SH in a dose and time-dependent manner. After administration of 1 to 10 mmol/L of methyl jasmonate for 24 h, the growth inhibition rate of cells reached 22.74% to 88.67%. After being treated by 2.5 mmol/L of methyl jasmonate, cis-jasmone and jasmonic acid, cell clone formation rate was decreased by 87.00% ,78.66% and 39.00% respectively. Cell cycles were arrested at S phase. Partial eells presented the characteristic morphological changes of apoptosis. The apoptotic rate induced by 1.5 mmol/L to 2.5 mmol/L methyl jasmonate for 24 h was 23.77% to 36. 43%. Conclusion Jasmonates could significantly inhibit the growth of SK-N-SH cells by suppressing proliferation and inducing ceil cycle arrest and apoptosis, which lays the groundwork for further investigation into the mechanisms of anti-tumor effects of jasmonates.