Department of Immunopathology, Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh 160012, India.
Department of Obstetrics & Gynecology, PGIMER, Chandigarh, India.
Gynecol Oncol. 2019 Oct;155(1):151-160. doi: 10.1016/j.ygyno.2019.07.023. Epub 2019 Jul 31.
Toll-like receptors constitute an important component of innate immune mechanism. HPV is a known etiological factor of cervical cancer and is known to interfere with the expression of TLRs and downstream signaling pathway. It remains poorly understood whether HPV modulates the expression of TLRs. Hence, understanding HPV mediated immune alterations might aid in identifying novel therapeutic targets. The aim was to study the relative gene expression of TLRs & downstream signaling pathway in cervical carcinoma.
Cervical squamous cell carcinoma (CSCC) and normal cervical tissues were obtained. Subsequent to HPV genotyping, mRNA expression profiling using PCR Array was performed. Protein expression of relevant genes with western blot was studied. Levels of cytokines in cervicovaginal washes were estimated using a Luminex multiplex platform.
All cases of cervical cancer were HR-HPV positive and predominant subtype was HPV16 (71.1%). Significant TLR4 upregulation and TLR2,7 downregulation were observed in HR-HPV infected cervix. TLR4,7 demonstrated low expression in CSCC. Molecules from cancer allied pathways; RELA, AKT, CDKN2A, and MDM2 demonstrated upregulation in CSCC. Protein expression data corroborated with gene expression profile. A diminished level of Th1 cytokines TNF-α, IFN-ɣ, IL-17, and IL-12 was observed in CSCC. Significantly increased levels of IL-1β, IL-6 and IL-2 were detected in HR-HPV infected cervix. Kaplan Meier curve demonstrated high TLR4 and low TLR7 expression was associated with poor prognosis.
The study demonstrates the HPV mediated dampening of the innate immune response in CSCC and provides support for exploring potential TLR2, 7 agonists as an adjunct therapy in CSCC patients.
Toll 样受体是先天免疫机制的重要组成部分。HPV 是宫颈癌的已知病因因子，已知其会干扰 TLRs 的表达及其下游信号通路。HPV 是否调节 TLRs 的表达仍知之甚少。因此，了解 HPV 介导的免疫改变可能有助于确定新的治疗靶点。本研究旨在研究宫颈癌中 TLRs 及下游信号通路的相对基因表达。
获取宫颈鳞状细胞癌（CSCC）和正常宫颈组织。进行 HPV 基因分型后，使用 PCR 阵列进行 mRNA 表达谱分析。通过 Western blot 研究相关基因的蛋白表达。使用 Luminex 多重平台估计宫颈阴道冲洗液中的细胞因子水平。
所有宫颈癌病例均为 HR-HPV 阳性，主要亚型为 HPV16（71.1%）。在 HR-HPV 感染的宫颈中观察到 TLR4 上调和 TLR2、7 下调。TLR4、7 在 CSCC 中表达水平较低。癌症相关途径的分子；RELA、AKT、CDKN2A 和 MDM2 在 CSCC 中表现出上调。蛋白表达数据与基因表达谱相符。在 CSCC 中观察到 Th1 细胞因子 TNF-α、IFN-γ、IL-17 和 IL-12 的水平降低。在 HR-HPV 感染的宫颈中检测到 IL-1β、IL-6 和 IL-2 的水平显著升高。Kaplan-Meier 曲线表明 TLR4 高表达和 TLR7 低表达与预后不良相关。
本研究表明 HPV 介导的 CSCC 先天免疫反应减弱，并为探索 TLR2、7 激动剂作为 CSCC 患者辅助治疗的潜力提供了支持。
