Aurora-A过表达通过激活NF-κBp65/ARPC4信号轴促进宫颈癌细胞的侵袭和转移

doi:10.12122/j.issn.1673-4254.2025.04.19

摘要/Abstract

摘要：

目的探索敲低肌动蛋白相关蛋白2/3复合体亚基4（ARPC4）对Aurora-A过表达诱导的宫颈癌细胞增殖、迁移、侵袭及上皮间质转化的影响以及Aurora-A表达调控ARPC4的分子机制。方法将pCDH-NC、pCDH-Aurora-A、pCDH-Aurora-A+shRNA-ARPC4、pCDH-Aurora-A质粒转染至Hela细胞中，并在第4组细胞中加入NF-κBp65抑制剂，RT-PCR检测转染效率。根据Aurora-A、ARPC4的表达情况及NF-κBp65通路的抑制状态将其分为4组：Vector组、Aurora-A过表达质粒组、Aurora-A过表达+ARPC4敲降组、Aurora-A过表达+NF-κBp65抑制剂组。EDU免疫荧光检测Hela细胞增殖情况；结晶紫染色检测Hela细胞细胞集落形成情况；划痕实验和Transwell实验分别检测Hela细胞迁移情况；Transwell基质胶检测Hela细胞侵袭情况；Western blotting检测Hela细胞上皮间充质化（EMT）情况、NF-κBp65及ARPC4的表达。结果 Aurora-A敲低的细胞中ARPC4表达下降，Aurora-A过表达的细胞中ARPC4表达上升（P<0.05）。过表达Aurora-A的宫颈癌细胞增殖、迁移及侵袭能力增强，而敲低ARPC4拮抗其作用（P<0.05）。Aurora-A过表达组NF-κBp65磷酸化水平增加，ARPC4表达水平增加（P<0.05）。Aurora-A可直接与NF-κBp65相互作用。与Aurora-A过表达组相比，Aurora-A过表达+NF-κBp65抑制剂组ARPC4表达下降（P<0.05）。结论 Aurora-A通过激活NF-κBp65信号通路上调ARPC4表达，促进宫颈癌细胞的迁移、侵袭和上皮间充质化的过程。

关键词: 宫颈癌, Aurora-A, 肌动蛋白相关蛋白2/3复合体亚基4, 增殖, 侵袭, 转移

Abstract:

Objective To investigate the regulatory effects of Aurora-A in regulating proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of cervical cancer cells and the role of actin-related protein 2/3 complex subunit 4 (ARPC4) in mediating its effects. Methods The plasmids pCDH-NC, pCDH-Aurora-A, and shRNA-ARPC4 were used for inducing Aurora-A overexpression or ARPC4 knockdown in HeLa cells. The cells were divided into vector group, Aurora-A overexpression group, Aurora-A overexpression+ARPC4 knockdown group, and Aurora-A overexpression+NF‑κBp65 inhibitor group and transfected with the corresponding plasmids. The proliferation, colony-forming ability, migration and invasion of the treated Hela cells was evaluated using EdU immunofluorescence assay, crystal violet staining, scratch assay, Transwell assay, and Matrigel assay. Western blotting was performed to detect the changes in cellular expressions of EMT-related proteins and expression levels of NF-κBp65 and ARPC4. Results The expression of ARPC4 was significantly decreased in HeLa cells with Aurora-A knockdown and increased in Aurora-A-overexpressing cells. Aurora-A overexpression obviously promoted proliferation, migration, and invasion abilities of HeLa cells, and these effects was significantly antagonized by ARPC4 knockdown. In Aurora-A-overexpressing cells, the phosphorylation level of NF-κBp65 and the expression level of ARPC4 were increased significantly, and application of the NF‑κBp65 inhibitor obviously lowered the expression level of ARPC4. Conclusion Aurora-A overexpression upregulates the expression of ARPC4 by activating the NF-κBp65 signaling pathway, thereby promoting migration, invasion and EMT of HeLa cells.

Key words: cervical cancer, Aurora-A, actin-related protein 2/3 complex subunit 4, proliferation, attack, transfer

岳雅清, 牟召霞, 王希波, 刘艳. Aurora-A过表达通过激活NF-κBp65/ARPC4信号轴促进宫颈癌细胞的侵袭和转移[J]. 南方医科大学学报, 2025, 45(4): 837-843.

Yaqing YUE, Zhaoxia MU, Xibo WANG, Yan LIU. Aurora-A overexpression promotes cervical cancer cell invasion and metastasis by activating the NF-κBp65/ARPC4 signaling axis[J]. Journal of Southern Medical University, 2025, 45(4): 837-843.

图/表 7

图1 敲低ARPC4拮抗Aurora-A过表达诱导的细胞增殖

Fig.1 ARPC4 knockdown antagonizes Aurora-A overexpression-induced enhancement of HeLa cell proliferation. A: EDU immunofluorescence staining for detecting cell proliferation in each group (scale bar=200 μm). B: Statistical chart of cell proliferation. *P<0.05.

图2 敲低ARPC4拮抗Aurora-A过表达诱导的细胞集落形成

Fig.2 ARPC4 knockdown antagonizes the effect of Aurora-A overexpression for promoting cell colony formation in HeLa cells. A: Colony formation in each group (crystal violet staining, scale bar=200 μm). B: Statistical chart of colony formation. *P<0.05 .

图3 敲低ARPC4拮抗Aurora-A过表达诱导的细胞迁移

Fig.3 ARPC4 knockdown antagonizes Aurora-A overexpression-induced enhancement of cell migration. A: Scratch assay for assessing migration ability of HeLa cells in each group (scale bar=200 μm). B: Transwell assay for assessing cell migration in each group (Crystal violet staining, scale bar=200 μm). C: Statistical chart of scratch assay in each group; D: Statistical chart of Transwell assay for each group. *P<0.05.

图4 敲低ARPC4拮抗Aurora-A过表达诱导的细胞侵袭

Fig.4 ARPC4 knockdown antagonizes the effect of Aurora-A overexpression for promoting cell invasion. A: Crystal violet staining for observing cell invasion in each group (scale bar=200 μm); B: Statistical chart of the invasive ability of each group. *P<0.05.

图5 敲低ARPC4拮抗Aurora-A过表达诱导的宫颈癌细胞EMT

Fig.5 ARPC4 knockdown antagonizes the effect of Aurora-A overexpression for promoting epithelial-mesenchymal transition of HeLa cells.

图6 过表达Aurora-A对NF-κBp65-ARPC4信号通路的影响

Fig.6 Effect of Aurora-A overexpression on NF-κBp65-ARPC4 signaling pathway in HeLa cells. A: Western blotting for detecting NF-κBp65-ARPC4 signaling pathway proteins in HeLa cells. B: Co-immunoprecipitation result showing interaction between Aurora-A and NF-κBp65.

图7 Aurora-A通过激活NF-κBp65信号通路上调ARPC4表达

Fig.7 Aurora-A upregulates ARPC4 expression by activating the NF‑κBp65 signaling pathway. The changes in ARPC4, NF‑κBp65, and p-NF‑κBp65 expressions were detected using Western blotting.

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