Research Paper|Volume 15, Issue 24|pp 14733—14748

Anoikis regulator GLI2 promotes NC cell immunity escape by TGF-β-mediated non-classic hedgehog signaling in colorectal cancer: based on artificial intelligence and big data analysis

Zhang Shanshan¹, Ding Fanfei², Sun Xuan¹, Lu Huina¹, Zhang Ye³, Li Jiayu³, Zhao Shuo¹, Pan Xue¹, Pu Yingye¹, Jin Chengjun¹, Pan Hang¹, Li Li¹

¹Laboratory Department of Changhai Hospital, First Affiliated Hospital of Naval Military Medical University, Shanghai, China
²Clinical Laboratory of PLA Naval Medical Center, Shanghai, China
³Medical Imaging Department of Changhai Hospital, First Affiliated Hospital of Naval Military Medical University, Shanghai, China

* Equal contribution

Received: July 5, 2023Accepted: October 18, 2023Published: December 29, 2023

https://doi.org/10.18632/aging.205283

Copyright: © 2023 Shanshan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Background: Anoikis is a speed-limited procedure to inhibit tumor metastasis during epithelial-mesenchymal transition (EMT). Previous studies have explored anoikis-related genes (ARG) in predicting prognosis and distinguishing tumoral immunity in many types of cancer. However, the role of ARGs in regulating NK cell exhaustion (NKE) and in predicting chemotherapy sensitivity is not clear. Therefore, it is necessary to work on it.

Methods: Gene expression profiles and clinical features are collected from TCGA and GEO, and data analysis is performed in R4.2.0.

Results: The ARGs-based no-supervised learning algorithm identifies three ARG subgroups, amongst which the prognosis is different. WCGNA and Artificial intelligence (AI) are applied to construct an NKE-related drug sensitivity stratification and prognosis identification model in digestive system cancer. Pathways association analysis screens out GLI2 is a key gene in regulating NKE by non-classic Hedgehog signaling (GLI2/TGF-β/IL6). In vitro experiments show that down-regulation of GLI2 enhances the CAPE-mediated cell toxicity and accompanies with down-regulation of PD-L1, tumor-derive IL6, and snial1 whereas the expression of cleaved caspas3, cleaved caspase4, cleaved PARP, and E-cadherin are up-regulated in colorectal cancer. Co-culture experiments show that GLI2- decreased colorectal tumor cells lead to down-regulation of TIM-3 and PD1 in NK cells, which are restored by TGF-bate active protein powder. Besides, the Elisa assay shows that GLI2-decreased colorectal tumor cells lead to up-regulation of IFN-gamma in NK cells.