IL-24对人宫颈癌HeLa细胞生物学行为的影响及其机制

doi:10.13481/j.1671-587X.20260417

摘要/Abstract

摘要：

目的探讨白细胞介素24（IL-24）调控V-Rel网状内皮增生病毒癌基因同源物B（RelB）对宫颈癌HeLa细胞生物学行为的影响，并阐明其作用机制。方法将HeLa细胞分为对照组［pcDNA3.1（+）组，转染pcDNA3.1（+）质粒］和IL-24过表达组［pcDNA3.1（+）-IL-24组，转染pcDNA3.1（+）-IL-24质粒］。采用实时荧光定量PCR（RT-qPCR）和Western blotting法检测2组细胞转染效率，细胞计数试剂盒8（CCK-8）法检测各组细胞增殖活性，流式细胞术检测2组细胞凋亡率，Transwell小室法检测2组细胞迁移率和细胞侵袭率，转录组测序（RNA-seq）法和生物信息学分析2组细胞差异表达基因，RT-qPCR和免疫荧光染色法检测IL-24过表达后HeLa细胞中RelB表达水平。验证实验，将HeLa细胞分为pcDNA3.1（+）组、pcDNA3.1（+）-IL-24组、HPV18 E6过表达组［pcDNA3.1（+）-IL-24+E6组，共转染pcDNA3.1（+）-IL-24质粒和pcDNA3.1（+）-HPV18 E6质粒］、HPV18 E7过表达组［pcDNA3.1（+）-IL-24+E7组，共转染pcDNA3.1（+）-IL-24质粒和pcDNA3.1（+）-HPV18 E7质粒］及HPV18 E6/E7过表达组［pcDNA3.1（+）-IL24+E6+E7组，共转染pcDNA3.1（+）-IL-24质粒、pcDNA3.1（+）-HPV18 E6质粒和pcDNA3.1（+）-HPV18 E7质粒］。RT-qPCR法检测人乳头瘤病毒18（HPV18）E6和HPV18 E7 mRNA表达水平及转染后HeLa细胞中RelB mRNA表达水平，Western blotting法检测HeLa细胞中RelB蛋白表达水平。结果与pcDNA3.1（+）组比较，pcDNA3.1（+）-IL-24组HeLa细胞中IL-24 mRNA和蛋白表达水平明显升高（P<0.01）；HeLa细胞增殖活性降低（P<0.001），细胞迁移率降低（P<0.05），细胞侵袭率降低（P<0.01），细胞凋亡率升高（P<0.01）。RNA-seq法共发现952个差异表达基因及与肿瘤发生发展相关的核因子κB（NF-κB）、环磷酸腺苷（cAMP）和磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）等多条信号通路。蛋白质-蛋白质相互作用（PPI）网络分析，RelB处于网络的核心调控区域。与pcDNA3.1（+）组比较，pcDNA3.1（+）-IL-24组HeLa细胞中HPV18 E6和HPV18 E7 mRNA表达水平降低（P<0.01），细胞中RelB mRNA表达水平明显降低（P<0.01）；过表达HPV18 E6和E7后，pcDNA3.1（+）-IL-24组HeLa细胞中RelB mRNA和蛋白表达水平升高（P<0.05）。结论 IL-24过表达可抑制HeLa细胞增殖、迁移和侵袭，并促进细胞凋亡，其机制可能与抑制HPV18 E6和E7表达、下调RelB蛋白表达水平有关联。

关键词: 宫颈肿瘤, 白细胞介素24, 转录组测序, V-Rel网状内皮增生病毒癌基因同源物B, 人乳头瘤病毒

Abstract:

Objective To discuss the effect of interleukin-24 （IL24） on the biological behaviors of cervical cancer HeLa cells through regulation of V-Rel reticuloendotheliosis viral oncogene homolog B （RelB）， and to clarify the mechanism. Methods The HeLa cells were divided into control group ［pcDNA3.1（+） group， transfected with pcDNA3.1（+） plasmid］ and IL-24 overexpression group ［pcDNA3.1（+）-IL-24 group，transfected with pcDNA3.1（+）-IL-24 plasmid］. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the transfection efficiencies of the cells in two groups； cell counting kit-8 （CCK-8） method was used to detect the proliferation activities of the cells in two groups； flow cytometry was used to detect the apoptotic rates of the cells in two groups； Transwell chamber assay was used to detect the migration rates and invasion rates of the cells in two groups； transcriptome sequencing （RNA-seq） method and bioinformatics were used to explore the differentially expressed genes in two groups of IL-24； RT-qPCR and immunofluorescence staining methods were used to detect the expression level of RelB mRNA and protein in HeLa cells after IL-24 overexpression. In validation experiment， the HeLa cells were divided into pcDNA3.1（+） group， pcDNA3.1（+）-IL-24 group， HPV18 E6 overexpression group ［pcDNA3.1（+）- IL-24+E6 group， co-transfected with pcDNA3.1（+）-IL-24 plasmid and pcDNA3.1（+）-HPV18 E6 plasmid］， HPV18 E7 overexpression group ［pcDNA3.1（+）-IL-24+E7 group， co-transfected with pcDNA3.1（+）-IL-24 plasmid and pcDNA3.1（+）-HPV18 E7 plasmid］， and HPV18 E6/E7 overexpression group ［pcDNA3.1（+）-IL-24+E6+E7 group， co-transfected with pcDNA3.1（+）-IL-24 plasmid， pcDNA3.1（+）- HPV18 E6 plasmid， and pcDNA3.1（+）-HPV18 E7 plasmid］.RT-qPCR method was used to detect the expression levels of human papillomavirus（HPV18） E6 and HPV18 E7 mRNA and the expression level of RelB mRNA in the HeLa cells after transfection； Western blotting method was used to detect the expression level of RelB protein in the HeLa cells. Results Compared with pcDNA3.1（+） group， the expression levels of IL-24 mRNA and protein in the HeLa cells in pcDNA3.1（+）-IL-24 group were significantly increased （P<0.01）， the proliferation activity of the HeLa cells was decreased （P<0.001）， the migration rate of cells was decreased （P<0.05）， the invasion rate of cells was decreased （P<0.01）， and the apoptotic rate was increased （P<0.01）. The RNA-seq results showed that a total of 952 differentially expressed genes were identified， which were involved in multiple signaling pathways related to tumor occurrence and development， including nuclear factor-kappa B （NF-κB）， cyclic adenosine monophosphate （cAMP）， and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt）. The protein-protein interaction （PPI） network analysis results showed that RelB was located in core regulatory region of the network. Compared with pcDNA3.1（+） group， the expression levels of HPV18 E6， HPV18 E7 and RelB mRNA in the HeLa cells in pcDNA3.1（+）-IL-24 group were significantly decreased（P<0.01）. Furthermore， after overexpression of HPV18 E6 and E7 in the HeLa cells in pcDNA3.1（+）-IL-24 group， the expression level of RelB mRNA and protein was significantly increased （P<0.05）. Conclusion Overexpression of IL-24 can inhibit the proliferation， migration and invasion of the HeLa cells and promote the cell apoptosis. The mechanism may be related to the inhibition of HPV18 E6 and E7 expressions and the down-regulation of RelB protein expression level.

Key words: Cervical neoplasm, Interleukin-24, Transcriptome sequencing, V-Rel reticuloendotheliosis viral oncogene homolog B, Human papillomaviru

中图分类号:

R737.33

王微,王靖,张杰,宋倩,孙甜甜,袁洪涛. IL-24对人宫颈癌HeLa细胞生物学行为的影响及其机制[J]. 吉林大学学报(医学版), 2026, 52(4): 1051-1061.

Wei WANG,Jing WANG,Jie ZHANG,Qian SONG,Tiantian SUN,Hongtao YUAN. Effect of IL-24 on biological behaviors of human cervical cancer HeLa cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2026, 52(4): 1051-1061.

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Gene	Primer sequence
IL-24	F：5'-AGATCTATGAGTTGGGGAC-3' R：5'-GAATTCTCAGAGATGGTAG-3'
RelB	F：5'-TAACAACCTGGGCATCCAGT-3' R：5'-CGCAGCTCTGATGTGTTTGT-3'
HPV18 E6	F：5'-GCGACCCTACAAGCTACCTG-3' R：5'-GTTGGAGTCGTTCCTGTCGT-3'
HPV18 E7	F：5'-AACATTTACCAGCCCGACGA-3' R：5'-TCGTCTGCTGAGCTTTCTAC-3'
β-actin	F：5'-CCCGAGCCGTGTTTCCT-3' R：5'-GTCCCAGTTGGTGACGATGC-3'