miR-147a Regulates Ferroptosis and Affects Invasion and Metastasis of Cervical Cancer Cells
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摘要:
目的 探讨miR-147a通过影响铁死亡过程对宫颈癌Hela细胞增殖和转移的调控作用。 方法 将Hela细胞孵育及传代,制备Hela单细胞悬液进行转染,转染后分为Control组、miR-147a mimics NC组、miR-147a mimics组、miR-147a inhibitor NC组、miR-147a inhibitor组、miR-147a mimics+ Ferrostatin-1 (1 μM)组。采用MTT检测细胞增殖率、Transwell检测细胞侵袭能力,划痕实验检测细胞的愈合能力,流式细胞术检测活性氧(reactive oxygen species,ROS)的表达情况,试剂盒检测亚铁离子(Fe2+)、铁离子(Fe3+)、乳过氧化物酶(Lactoperoxidase,LPO)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)、谷胱甘肽(glutathione,GSH)的表达变化。 结果 miR-147a mimics组的细胞增殖率、细胞侵袭数量和细胞迁移距离较Control组降低(P < 0.05);miR-147a inhibitor组的细胞增殖率、细胞侵袭数量和细胞迁移距离较Control组升高(P < 0.05)。较Control组,miR-147a mimics组的Fe2+、LPO、ROS的表达升高,Fe3+、GSH-PX和GSH的表达降低(P < 0.05)。miR-147a inhibitor组的Fe2+、LPO、ROS的表达降低,Fe3+、GSH-PX和GSH的表达升高(P < 0.05)。miR-147a mimics+Ferrostatin-1组的细胞增殖率、细胞侵袭数量和细胞迁移距离较miR-147a mimics组升高(P < 0.05)。与miR-147a mimics组比较,miR-147a mimics+Ferrostatin-1组的Fe2+、LPO、ROS的表达水平降低,(P < 0.05);Fe3+、GSH-PX和GSH的表达水平升高(P < 0.05)。 结论 miR-147a可以促进铁死亡,抑制宫颈癌细胞的增殖和转移。 Abstract:Objective To investigate the regulatory role of miR-147a in affecting cell proliferation and metastasis of cervical cancer Hela cells through ferroptosis. Methods Hela cells were cultured and passaged, and then a Hela single-cell suspension was prepared and transfected. After transfection, the cells were divided into control group, miR-147a mimics NC group, miR-147a mimics group, miR-147a inhibitor NC group, miR-147a inhibitor group, and miR-147a mimics+Ferrostatin-1 (1 μM) group. MTT assay was used to detect cell proliferation, Transwell assay to assess cell invasion capacity, scratch test to evaluate cell healing ability, flow cytometry to detect reactive oxygen species (ROS) expression, and assay kits to measure expression changes of ferrous ions (Fe2+), ferric ions (Fe3+), lactoperoxidase (LPO), glutathione peroxidase (GSH-PX), and glutathione (GSH). Results Compared to the control group, the miR-147a mimics group showed reduced cell proliferation rate, cell invasion quantity, and cell migration distance (P < 0.05); the miR-147a inhibitor group exhibited increased cell proliferation rate, cell invasion quantity, and cell migration distance (P < 0.05). Compared to the control group, the miR-147a mimics group showed increased Fe2+, LPO, and ROS expression, with decreased Fe3+, GSH-PX, and GSH expression (P < 0.05). The miR-147a inhibitor group showed decreased Fe2+ , LPO, and ROS expression, with increased Fe3+, GSH-PX, and GSH expression(P < 0.05). The miR-147a mimics + Ferrostatin-1 group exhibited increased cell proliferation rate, cell invasion quantity, and cell migration distance compared to the miR-147a mimics group (P < 0.05). Compared to the miR-147a mimics group, the miR-147a mimics + Ferrostatin-1 group showed decreased Fe2+, LPO, and ROS expression levels (P < 0.05), with increased Fe3+, GSH-PX, and GSH expression levels (P < 0.05). Conclusion miR-147a can promote ferroptosis and inhibit the proliferation and metastasis of cervical cancer cells. -
Key words:
- Cervical cancer /
- miR-147a /
- Ferroptosis /
- Metastasis
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表 1 MTT检测Hela细胞增殖情况[($ \bar x \pm s $),n = 3]
Table 1. Detection of the proliferation of Hela cells by MTT[($ \bar x \pm s $),n = 3]
组别 Hela细胞增殖活性(%) Control组 100.00±0.02 miR-147a mimics NC组 99.95±4.55 miR-147a mimics组 85.36±3.76### miR-147a inhibitor NC组 100.12±1.37 miR-147a inhibitor组 114.39±2.25### F 37.782 P <0.001*** ***P < 0.05;与Control组比较,###P < 0.001。 表 2 不同分组细胞的迁移和侵袭情况[($ \bar x \pm s $),n = 3]
Table 2. Migration and invasion of cells in different groups[($ \bar x \pm s $),n = 3]
组别 细胞迁移距离(%) 细胞侵袭数 Control组 42.12±3.19 52.33±4.04 miR-147a mimics NC组 41.94±4.42 51.33±3.21 miR-147a mimics组 29.13±1.41### 32.67±3.05### miR-147a inhibitor NC组 43.46±7.35 52.00±5.57 miR-147a inhibitor组 55.32±1.27### 72.67±4.73### F 14.769 33.634 P <0.001*** <0.001*** ***P < 0.05;与Control组比较,###P < 0.001。 -
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