阴道微环境中IL-10通过JAK1/TYK2/STAT3通路对宫颈病变进展机制的研究

The mechanism of IL-10 in the vaginal microenvironment affecting the progression of cervical lesions through JAK1/TYK2/STAT3 pathway

目的:探讨IL-10在不同宫颈病变患者阴道灌洗液及宫颈组织中的表达,检测上调IL-10对正常宫颈上皮细胞增殖、迁移、侵袭功能及JAK1/TYK2/STAT3通路因子表达水平的影响,研究IL-10影响宫颈病变的可能机制。方法:ELISA法及免疫组化法检测正常宫颈及宫颈病变患者阴道灌洗液和宫颈组织中IL-10的表达情况。分别将IL-10过表达质粒(pcDNA3.1-IL-10)及空载体质粒转染至HcerEpic细胞,采用qRT-PCR技术检测转染后mRNA水平的变化,通过CCK-8实验、平板划痕实验和Transwell小室实验分析IL-10对正常宫颈细胞增殖、迁移和侵袭的影响,Western blot法检测通路因子JAK1、TYK2、STAT3及下游因子MMP-9水平的变化。结果:IL-10在正常宫颈、低级别鳞状上皮内病变、高级别鳞状上皮内病变及宫颈鳞癌患者的阴道灌洗液及组织中表达水平逐步增高。过表达IL-10的正常宫颈上皮细胞较对照组增殖率上升,迁移及侵袭能力增强。IL-10过表达组中JAK1、TYK2、STAT3及MMP-9较对照组表达升高。加JAK1/TYK2抑制剂PF06700841后,宫颈上皮细胞增殖及侵袭转移能力较IL-10过表达组降低,STAT3、MMP-9表达水平下降。结论:阴道微环境中IL-10可能通过激活JAK1/TYK2/STAT3通路促进正常宫颈上皮细胞增殖,并增加其迁移及侵袭能力。

Objective:To investigate the expression of interleukin 10(IL-10)in patients with different cervical lesions in their vaginal lavage fluid and cervical tissues,and to explore its effect and the possible mechanisms on the proliferation,migration and invasion of normal cervical epithelial cells(HcerEpic).Methods:Enzyme-Linked Immunosorbent Assay(ELISA)and the immunohistochemical Elivision two-step was used to detect the expression of IL-10 in vaginal lavage fluid and cervical tissues of control group and patients with cervical lesions.IL-10 overexpression plasmid and empty plasmid were transfected into HcerEpic respectively,the expression level was detected using real-time fluorescent quantitative PCR.The proliferation was detected by CCK-8 test.Wound healing assay and Transwell migration assay were used to evaluate the migration of the cell lines.The expression levels of JAK1、TYK2、STAT3 and MMP-9 proteins were detected by Western blot.Results:The expression of IL-10 in vaginal lavage fluid and cervical tissues increased gradually with the severity of cervical lesions.CCK-8 assay showed that IL-10 promoted the growth of the cells in a time-dependent manner.Also,it effectively improved migration and invasion of the cells.At the same time,the cells showed significantly up-regulation of JAK1、TYK2、STAT3 and MMP-9 after IL-10 gene transfected into it.After adding JAK1/TYK2 inhibitor PF06700841 to the normal cervical epithelial cells,the proliferation,invasion and metastasis ability of the cells were decreased,and the expression levels of STAT3 and MMP-9 were decreased compared with IL-10 overexpression group.Conclusions:IL-10 can promote cervical cell growth,improve migration and invasion possibly by activating JAK1/TYK2/STAT3 pathway.