医学分子生物学杂志 ›› 2022, Vol. 19 ›› Issue (3): 219-224.doi: 10.3870/j.issn.1672-8009.2022.03.007

• 论著 • 上一篇    下一篇

舒芬太尼诱导宫颈癌细胞自噬和凋亡并抑制癌细胞恶性增殖

  

  1. 1河北北方学院附属第一医院放疗科 河北省张家口市, 075000  2北京中医药大学第三附属医院放疗科 北京市, 100029
  • 出版日期:2022-05-31 发布日期:2022-06-20
  • 基金资助:
    张家口市 2021 年市级科技计划自筹经费项目 (No. 2121192H)

Sufentanil Induces Autophagy and Apoptosis of Cervical Cancer Cells and Inhibits Their Proliferation

  1. 1Department of Radiotherapy, the First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei, 075000, China  2Department of Radiotherapy, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing, 100029, China 
  • Online:2022-05-31 Published:2022-06-20

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摘要： 目的 探讨舒芬太尼对宫颈癌细胞 SiHa 自噬、 凋亡及细胞增殖的影响。 方法 采用 CCK8 法检 测舒芬太尼梯度浓度 (3. 125、 6. 25、 12. 5、 25、 50、 100、 200、 400、 800 nmol / L) 作用下宫颈癌细胞 SiHa 细胞活力, 选取 4 个舒芬太尼作用浓度 (0、 25、 50、 100 nmol / L) 处理 SiHa 细胞 24 h, 采用流式细胞法、 克隆形成实验检测 SiHa 细胞凋亡及增殖情况, 免疫荧光法检测各组细胞 LC3 水平, 采用 Western 印迹检测 自噬相关蛋白 LC3Ⅱ/ LC3Ⅰ、 Beclin1、 ATG7 与增殖、 凋亡相关蛋白 P21、 Survivin 水平, 采用 RT-PCR 检测 细胞增殖相关基因 Ki67、 PCNA 表达水平。 结果 随着舒芬太尼处理浓度的增加, SiHa 细胞活力逐渐降低。 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞的 LC3Ⅱ/ LC3Ⅰ、 Beclin1、 ATG7 水平显著高于 0 nmol / L 舒芬太 尼处理, 免疫荧光检测结果显示 50、 100 nmol / L 舒芬太尼处理下 LC3 荧光信号高于 0 nmol / L 舒芬太尼处 理, 流式细胞仪检测结果显示 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞凋亡高于 0 nmol / L 舒芬太尼处理。 克隆形成实验结果显示 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞克隆形成率降低, Survivin 水平与 Ki67、 PCNA 表达水平降低, 而 P21 水平升高。 结论 舒芬太尼对宫颈癌细胞自噬、 凋亡有促进作用, 对细胞增 殖有抑制作用, 可能与调节细胞自噬、 凋亡、 增殖相关蛋白或基因水平有关。 

关键词: 宫颈癌, 舒芬太尼, 细胞自噬, 细胞凋亡, 细胞增殖

Abstract: Objective To explore the effects of sufentanil on autophagy, apoptosis and proliferation of cervical cancer cells SiHa. Methods SiHa cells were treated with sufentanil (3. 125, 6. 25, 12. 5, 25, 50, 100, 200, 400, 800 nmol / L ) . Cell viability was measured by CCK8. Four concentrations of sufentanil (0, 25, 50, 100 nmol / L) were then selected to treat SiHa cells for 24 h for the following experiments. The apoptosis and proliferation of SiHa cells were measured by flow cytometry and colony formation assay. The level of LC3 in each group was measured by immunofluorescence method. The levels of autophagy-related proteins (LC3Ⅱ/ LC3Ⅰ, Beclin1, ATG7 ), proliferation-and apoptosis-related proteins ( P21, Survivin ) were detected by Western blotting. The expression levels of cell proliferation-related genes (Ki67, PCNA) were detected by RT-PCR. Results The viability of SiHa cells decreased with the increased concentration of sufentanil and the effect was in a dose-dependent manner. The expression levels of LC3Ⅱ/ LC3 Ⅰ, Beclin1 and ATG7 in SiHa cells treated with 50 nmol / L and 100 nmol / L sufentanil were signif- icantly higher than those treated with 0 nmol / L sufentanil. The immunofluorescence method showed that the fluorescence signal of LC3 in SiHa cells treated with 50 and 100 nmol / L sufentanil was higher than that in cells treated with 0 nmol / L sufentanil. Flow cytometry showed that the apoptosis rate of SiHa cells treated with 50 and 100 nmol / L sufentanil was higher than that of cells treated with 0 nmol / L sufentanil. Clone formation assay showed that the clonal formation rate of SiHa cells was decreased after 50 and 100 nmol / L sufentanil treatment. The expression levels of Survivin, Ki67 and PCNA were decreased, and the expression level of P21 was increased. Conclusion Sufentanil can promote autophagy and apoptosis of cervical cancer cells, and inhibit their proliferation, which may be through the regulation of autophagy-, apoptosis- and proliferation-related proteins or genes.

Key words: cervical cancer, sufentanil, cell autophagy, apoptosis, cell proliferation

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