目的 研究吉马酮诱导肺癌 A549、 鼻咽癌 CNE-1、 肝癌 HepG2 细胞凋亡的机制 。 方法 50、 150、 200、250、300 μmol·L^-1的吉马酮处理肝癌 HepG2、肺癌 A549、鼻咽癌 CNE-1、结肠癌 Caco-2 细胞 24、48、72 h 后，MTT 实验检测细胞的存活率的变化。100、150、200 μmol·L^-1的吉马酮分别处理 A549、HepG2、CNE-1细胞 48 h后采用流式细胞术检测细胞凋亡的变化；Western blotting 实验检测细胞凋亡标志蛋白 cleaved-Caspase-3、Caspase-3 的变化，检测乙型肝炎 X相互作用蛋白（HBXIP）、p53蛋白的表达量变化。分别在 A549、HepG2、CNE-1细胞中应用 siRNA 敲低 HBXIP，Westernblotting实验检测 HBXIP的敲低效果及 p53蛋白表达变化；MTT实验检测敲低 HBXIP对吉马酮诱导的细胞增殖抑制作用的影响。结果 吉马酮对鼻咽癌CNE-1、肝癌HepG2细胞增殖抑制作用较强，与溶剂对照组比较，200、250、300 μmol·L^-1组细胞存活率显著下降（P＜0.05）；在高浓度时对肺癌A549细胞增殖抑制效果较强，与溶剂对照组比较，250、300 μmol·L^-1组细胞存活率显著下降（P＜0.05）；对结肠癌Caco-2细胞作用相对较弱。与对照组比较，100、150、200 μmol·L^-1吉马酮处理A549、HepG2、CNE-1细胞 48 h后，凋亡率显著升高（P＜0.05），cleaved-Caspase-3、p53蛋白表达显著上升（P＜0.05）；以 150、200 μmol·L^-1吉马酮处理A549、HepG2、CNE-1细胞48 h后，HBXIP的蛋白表达显著降低（P＜0.05）。siRNA-HBXIP处理48 h后，与对照组比较，HBXIP的蛋白表达量显著下降（P＜0.05），p53蛋白表达量显著上升（P＜0.05）。与单独使用的相同浓度的吉马酮相比，沉默HBXIP后A549、HepG2、CNE-1细胞对吉马酮的敏感度明显提高，150、200、250 μmol·L^-1组均差异显著（P＜0.05）。结论 吉马酮可以抑制A549、HepG2、CNE-1细胞增殖、诱导细胞凋亡，作用机制可能与调控HBXIP/p53信号通路相关。

Objective Study the mechanism of gemmazone induced apoptosis in A549, CNE-1, and HepG2 cells.Methods After 50, 150, 200, 250, and 300 μmol·L^-1 germacrone treated in HepG2, A549, CNE-1, and Caco-2 cells for 24, 48 and 72 h, the MTT assay was preformed to detect the changes of cell viability. When 100, 150 and 200 μmol·L^-1 germacrone treated in A549, HepG2 and CNE-1 cells for 48 h, the changes of cell apoptosis were detected by flow cytometry, and the Western blotting was used to detect the changes of cleaved-Caspase 3, Caspase-3 HBXIP, and P53 proteins. The expressions of HBXIP and P53 were detected by Western blotting after HBXIP knocked down in A549, HepG2 and CNE-1 cells. The effect of knockdown of HBXIP on the inhibitory proliferation induced by germacrone was detected by MTT assay.Results Gemmazone had a strong inhibitory effect on the proliferation of nasopharyngeal carcinoma CNE-1 and liver cancer HepG2 cells, compared with the solvent control group, the cell survival rate of the 200, 250, and 300 μmol·L^-1 group significantly decreased (P < 0.05). At high concentrations, the inhibitory effect on the proliferation of lung cancer A549 cells was better, compared with the solvent control group, the cell survival rate of the 250, 300 μmol·L^-1 group significantly decreased (P < 0.05). The effect on colon cancer Caco-2 cells was relatively weak. Compared with the control group, after 48 hours of treatment with 100, 150, 200 μmol·L^-1 gemmazone, the apoptosis rate of A549, HepG2, and CNE-1 cells significantly increased (P < 0.05), and the expression of cleaved-Caspase-3 and p53 proteins significantly increased (P < 0.05). After 48 hours of treatment with 150, 200 μmol·L^-1 gemmazone in A549, HepG2, and CNE-1 cells, the protein expression of HBXIP was significantly reduced (P < 0.05). After 48 hours of siRNA-HBXIP treatment, compared with the control group, the protein expression of HBXIP significantly decreased (P < 0.05) and the expression of p53 protein significantly increased (P < 0.05). Compared with the same concentration of gemmazone used alone, silencing HBXIP significantly increased the sensitivity of A549, HepG2, and CNE-1 cells to gemmazone, with values of 150, 200, and 250 μmol·L^-1 group showed significant differences (P < 0.05).Conclusion Gematone can inhibit the proliferation of A549, HepG2, and CNE-1 cells and induce cell apoptosis, and the mechanism of action may be related to the regulation of the HBXIP/p53 signaling pathway.

R285.5

合肥市自然科学基金（2021012）；安徽省高等学校科学研究项目（自然科学类）（2022AH050712）；合肥市第二人民医院院级课题（2021ygkt35）