混合血统激酶4（MLK4）是丝氨酸/苏氨酸激酶混合血统激酶（MLKs）家族的成员。针对子宫颈鳞状细胞癌和子宫颈腺癌（CESC）中与免疫相关的靶标的报道很少，而MLK4在子宫颈癌中的作用仍需研究。使用包含306个CESC组织和3个癌旁组织样本的TCGA数据库分析了CESC中MLK4的表达，并使用Deseq2软件包（Benjamini-Hochberg校正后p值<0.05且log2倍数差≥|2|）评估了MLK4对免疫侵袭的影响。组织微阵列用于验证CESC患者中MLK4的表达，结果发现MLK4在CESC中明显过表达，并与WHO分级密切相关。多种分析算法揭示高表达的MLK4与CESC中免疫细胞浸润呈负相关。分析显示，MLK4表达与包括CD8+T细胞在内的多种免疫细胞浸润呈负相关，而MLK4 mRNA表达与免疫检查点PD-L1、CTLA4、LAG3呈正相关，与免疫促进基因CD86和CD80呈负相关。此外，采用离体实验研究了MLK4在C33A细胞中的生物学特性。EDU和Transwell实验证明，降低C33A细胞中MLK4表达导致细胞增殖和侵袭的减少。MLK4沉默导致炎性细胞因子IL-1β（p<0.05）、TNF-α（p<0.01）和IL-6（p<0.05）表达显著增加。细胞实验结果表明，敲低MLK4会抑制已建立的生化标志物CEA、AFP和HCG的表达。因此，可以推断MLK4可能对宫颈癌的发展和进展产生显著影响。 版权：© 2023 Gong等。本文为开放获取文章，根据创作公共许可证（Creative Commons Attribution License）可以在任何媒体中自由使用、分发和复制，只要原作者和来源得到适当的署名。

Mixed pedigree kinase 4 (MLK4) is a member of the serine/threonine kinases mixed pedigree kinase (MLKs) family. Few reports on immune-related targets in Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), and the role of MLK4 in cervical cancer remains to be studied. The expression of MLK4 in CESC was analyzed by TCGA database containing 306 CESC tissues and 3 peritumoral tissue samples, and the effect of MLK4 on immune invasion was evaluated using the Deseq2 package(Benjamini-Hochberg corrected p-value < 0.05 and log2 fold change ≥|2|). Tissue microarray was used to verify the expression of MLK4 in CESC patients, and it was found that MLK4 was significantly overexpressed in CESC, and significantly correlated with WHO grade. Multiple analysis algorithms revealed that the high expression of MLK4 was negatively correlated with immune cell infiltration in CESC. Analysis showed that MLK4 expression was negatively correlated with the infiltration of various immune cells including CD8+T cells, and MLK4 mRNA expression was positively correlated with immune checkpoints PD-L1,CTLA4, LAG3, and negatively correlated with immune promotion genes CD86 and CD80. Furthermore, vitro assays were performed to investigate the biological characteristics of MLK4 in C33A cells. The EDU and transwell assays demonstrated that the decrease in MLK4 expression in C33A cells resulted in a decrease in cell proliferation and invasion. The silencing of MLK4 resulted in a significant increase in the expression of inflammatory cytokines IL-1β(p<0.05), TNF-α(p<0.01), and IL-6 (p<0.05). The results of cell assays indicate that knocking down MLK4 would inhibit the expression of established biochemical markers CEA, AFP and HCG. Hence, it is plausible that MLK4 could potentially exert a significant influence on the development and progression of Cervical cancer.Copyright: © 2023 Gong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.