南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (11): 1280-1286.doi: 10.12122/j.issn.1673-4254.2019.11.03

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高尔基体磷蛋白3通过促进细胞自噬抑制紫杉醇诱导的hela细胞凋亡

王镇南，郑玉菡，黄海丽，何惠娟，贾庆明   

  • 出版日期:2019-12-05 发布日期:2019-11-20
  • 基金资助:

Golgi phosphoprotein 3 overexpression inhibits paclitaxel-induced apoptosis in HeLa cells by promoting autophagy

  • Online:2019-12-05 Published:2019-11-20

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摘要： 目的 探讨紫杉醇处理hela细胞时，高尔基体磷蛋白3（Golph3）对细胞自噬的调控和对紫杉醇引起的细胞凋亡的影响。方法 通过慢病毒感染的方法上调hela细胞中Golph3的表达；通过转染siRNA方法下调hela细胞中Golph3的表达；使用透射电镜观察细胞中的自噬小体；免疫印记检测自噬标记物LC3II的蛋白表达水平；通过流式细胞仪检测细胞凋亡率；使用自噬抑制剂 3-MA抑制 hela细胞自噬。结果 Golph3慢病毒可上调 hela细胞 Golph3蛋白表达，siRNA可抑制 hela细胞 Golph3蛋白表达。透射电镜观察结果显示，Golph3过表达组细胞自噬小体的数量增多，siRNA组细胞自噬小体的数量减少。免疫印迹结果显示，Golph3组细胞中自噬标记物LC3II表达增强，p62表达减弱；siRNA组LC3II表达抑制，p62表达增强。凋亡检测结果显示，Golph3组紫杉醇诱导的细胞凋亡率下降（P<0.01），siRNA组细胞凋亡率上升（P<0.01）。自噬抑制剂3-MA单独处理不会显著增加hela细胞凋亡率。3-MA处理同时下调Golph3表达可以促进紫杉醇引起的细胞凋亡（P<0.01）。结论 紫杉醇处理hela细胞时，上调Golph3促进hela细胞自噬，抑制细胞凋亡；下调Golph3抑制hela细胞自噬，促进细胞凋亡。Golph3通过调控细胞自噬影响紫杉醇引起的hela细胞凋亡。

Abstract: Objective To investigate the effect of Golgi phosphoprotein 3 (Golph3) on paclitaxel- induced apoptosis and autophagy in HeLa cells. Methods HeLa cells were transfected with a lentiviral vector expressing Golph3 or a small interfering RNA (siRNA) targeting Golph3 for up-regulation or down-regulation of Golph3 which was verified by Western blotting. The autophagic bodies in the cells were observed using transmission electron microscopy. The expression of autophagy markers p62 and LC3 were detected using Western blotting, and the cell apoptosis was examined by PI/Anexin V-FITC double staining and flow cytometry. The effects of blocking autophagy was evaluated by treatment of the cells with the autophagy inhibitor 3-MA. Results Transmission electron microscopy showed that the lentivirus-mediated overexpression of Golph3 significantly increased the number of autophagic bodies and interference of Golph3 expression significantly decreased autophagic bodies in HeLa cells. Western blotting showed that Golph3 overexpression caused an increased expression of LC3 and decreased the accumulation of p62 in the cells, and interference of Golph3 resulted in the reverse changes. The cell apoptosis induced by paclitaxel was significantly decreased in Golph3-overexpressing HeLa cells and increased in the cells with Golph3 knockdown (P<0.01). Treatment with 3-MA alone did not obviously affect HeLa cell apoptosis, but in cells with Golph3 knockdown, 3-MA significantly enhanced paclitaxel-induced apoptosis (P<0.01). Conclusion Up-regulation of Golph3 promotes autophagy and inhibits paclitaxel-induced apoptosis, whereas suppression of Golph3 inhibits autophagy and enhances paclitaxel- induced apoptosis in HeLa cells. Keywords: paclitaxel; chemotherapy; autophagy; Golgi phosphoprotein 3