[1]杨加宁,张立然.雷帕霉素抑制mTOR激活自噬并调控铁死亡降低宫颈癌细胞增殖、侵袭及迁移能力的实验研究[J].现代检验医学杂志,2025,40(03):42-46.[doi:10.3969/j.issn.1671-7414.2025.03.008]
　YANG Jianing,ZHANG Liran.Experimental Study of Rapamycin Inhibiting mTOR Activation Autophagy and Regulating Ferroptosis to Reduce the Proliferation, Invasion and Migration of Cervical Cancer Cells[J].Journal of Modern Laboratory Medicine,2025,40(03):42-46.[doi:10.3969/j.issn.1671-7414.2025.03.008]
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雷帕霉素抑制mTOR激活自噬并调控铁死亡降低宫颈癌细胞增殖、侵袭及迁移能力的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年03期
页码:
42-46
栏目:
论著
出版日期:
2025-05-15

文章信息/Info

Title:
Experimental Study of Rapamycin Inhibiting mTOR Activation Autophagy and Regulating Ferroptosis to Reduce the Proliferation, Invasion and Migration of Cervical Cancer Cells
文章编号:
1671-7414（2025）03-042-05
作者:
杨加宁，张立然
（黑龙江省中医药科学院妇科，哈尔滨 150080）
Author(s):
YANG Jianing，ZHANG Liran
（Department of Gynecology，Heilongjiang University of Traditional Chinese Medicine, Harbin 150080，China）
关键词:
宫颈癌; 哺乳动物雷帕霉素靶蛋白; 自噬; 铁死亡; 增殖; 迁移; 侵袭
分类号:
R737.33；R730.43
DOI:
10.3969/j.issn.1671-7414.2025.03.008
文献标志码:
A
摘要:
目的探究雷帕霉素抑制哺乳动物雷帕霉素靶蛋白（mTOR）激活自噬并调控铁死亡对宫颈癌细胞增殖、侵袭及迁移能力的影响及可能机制。方法培养正常宫颈上皮细胞H8，宫颈癌细胞Caski，将Caski细胞分组为宫颈癌组、雷帕霉素组与Erastin组。Westernblotting检测细胞中mTOR，Beclin1，微管相关蛋白轻链3II（LC3Ⅱ）、溶质载体家族7成员11（SLC7A11）和谷胱甘肽过氧化物酶4（GPX4）蛋白水平。RT-qPCR检测细胞中mTOR，Beclin1，LC3Ⅱ，SLC7A11和GPX4mRNA水平。相应试剂盒检测细胞中活性氧（ROS）、谷胱甘肽（GSH）和Fe2+水平。平板克隆检测细胞克隆能力；划痕实验检测细胞迁移能力；Transwell实验检测细胞侵袭能力。结果与H8细胞相比，Caski细胞中mTOR，SLC7A11和GPX4蛋白表达增加（t=10.58，36.66，14.68），Beclin1和LC3Ⅱ蛋白表达减少（t=23.00，9.50），差异具有统计学意义（均P<0.05）。与宫颈癌组相比，雷帕霉素组mTOR蛋白及mRNA表达减少（t=25.00，12.50），Beclin1，LC3Ⅱ蛋白及mRNA表达增加（t=6.84～30.31），差异具有统计学意义（均P<0.05）；GSH水平减少（t=9.15），ROS及Fe2+水平增加（t=7.64，6.81），细胞增殖能力、迁移能力及侵袭能力降低（t=19.03，8.69，23.00），差异具有统计学意义（均P<0.05）；Erastin组Caski细胞的增殖能力、迁移能力及侵袭能力均降低，差异具有统计学意义（t=25.34，4.72，6.43，均P<0.05）。结论雷帕霉素可通过抑制mTOR激活自噬并调控SLC7A11/GPX4通路介导的铁死亡降低宫颈癌细胞的增殖、侵袭及迁移能力。
Abstract:
Objective To explore the effect and possible mechanism of rapamycin inhibiting mammalian target of rapamycin (mTOR) activation autophagy and regulating iron death on proliferation, invasion and migration of cervical cancer cells. Methods Normal cervical epithelial cells H8 and cervical cancer cells Caski were cultured and divided into H8 and Caski. Caski cells were further cultured and divided into cervical cancer, rapamycin and Erastin groups. Western blotting detected the protein levels of mTOR, Beclin1, microtubule-associated protein light chain 3 II (LC3Ⅱ), recombinant solute carrier family 7, member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) in the cells. The mRNA levels of mTOR, Beclin1, LC3Ⅱ, SLC7A11 and GPX4 were detected by RT-qPCR. The kit detected reactive oxygen species (ROS), glutathione (GSH) and Fe2+ levels in cells. Plate cloning was used to detect the cloning ability of cells. Cell migration ability was detected by scratch test. Transwell assay was used to detect cell invasion ability. Results Compared with H8 cells, the protein expressions of mTOR, SLC7A11, GPX4 were increased in Caski cells (t=10.58, 36.66, 14.68). Beclin1, LC3 Ⅱ decreased protein expression (t=23.00, 9.50), and the differences were statistically significant (all P<0.05). Compared with the cervical cancer group, the expressions of mTOR protein and mRNA in the rapamycin group were decreased (t=25.00, 12.50), the expressions of Beclin1, LC3Ⅱ protein and mRNA were increased (t=6.84 ～ 30.31), and the differences were statistically significant (all P<0.05). The levels of GSH were decreased (t=9.15), ROS and Fe2+ were increased (t=7.64, 6.81), and the cell proliferation, migration and invasion ability were decreased (t=19.03, 8.69, 23.00), and the differences were statistically significant (all P<0.05). The proliferation ability，migration ability, and invasion ability of Caski cells in the Erastin group were decreased, and the differences were statistically significant (t=25.34, 4.72, 6.43, all P<0.05). Conclusion Rapamycin can reduce the proliferation, invasion and migration of cervical cancer cells by inhibiting mTOR activation of autophagy and regulating ferroptosis mediated by SLC7A11/GPX4 pathway.

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备注/Memo

备注/Memo:
基金项目：黑龙江省中医药科研项目（项目编号：ZHY2023-082）。
作者简介：杨加宁（1999-），女，硕士，研究方向：中医、妇科、针灸，E-mail：wenwenwang890909@163.com。
更新日期/Last Update: 2025-05-15