Lin Xuyang, Han Xiao, Zhou Wubi, Gong Xiaoxia, Zhou Yu, Wang Qilong, Zhang Chengwan
Department of Stomatology, The Affiliated Huai'an No. 1 People's Hospital of Nanjing Medical University Huai'an 223001, Jiangsu, China.
State Key Laboratory Cultivation Base of Research, Prevention and Treatment for Oral Diseases, Nanjing Medical University Nanjing 210000, Jiangsu, China.
Am J Cancer Res. 2024 Nov 15;14(11):5486-5503. doi: 10.62347/IDCP2061. eCollection 2024.
Collagen, a primary protein component of the extracellular matrix (ECM), undergoes a notable series of alterations concomitant with the growth of the tumor. Procollagen-lysine,2-oxoglutarate 5-dioxygenase 3 (PLOD3) is involved in the synthesis of collagen and has been associated with a variety of cancers. However, it is unclear how PLOD3 functions in esophageal squamous cell carcinoma (ESCC).
Differentially expressed genes between ESCC and adjacent normal tissues were identified using proteomic and transcriptomic analyses. These genes were then subjected to survival analysis to identify prognostic markers. Immune cell infiltration in the two subgroups was evaluated. Spearman's correlation analysis was performed to examine the correlation between PLOD3 and RBM15 expression in TCGA-ESCC database. shRNA-mediated approach was used to knockdown RBM15 in ESCC cells. The effects of RBM15 knockdown on PLOD3 expression were assessed by real-time PCR and Western blot. Moreover, COX algorithm was employed to construct a prognostic signature.
PLOD3 was found to be highly expressed in ESCC patients and correlated with a favorable prognosis. Immune cell infiltration estimation indicated tumor-infiltrating CD4+ T cell was increased in PLOD3-high group. Correlation analysis revealed that PLOD3 was associated with RBM15 and was closely related to CD4+ T cell infiltration. Moreover, loss-of-function approaches showed that depletion of RBM15 attenuated PLOD3 expression in ESCC cells. Following univariate and multivariate Cox regression analyses, PLOD3 and RBM15 were identified as a two-gene prognostic signature for ESCC.
RBM15 enhances tumor-infiltrating CD4+ T Cell abundance in ESCC by regulating PLOD3. Two new independent prognostic factors, PLOD3 and RBM15, may be useful in predicting the prognosis of ESCC.
胶原蛋白是细胞外基质（ECM）的主要蛋白质成分，伴随肿瘤生长会发生一系列显著变化。前胶原赖氨酸2-氧戊二酸5-双加氧酶3（PLOD3）参与胶原蛋白的合成，并与多种癌症相关。然而，PLOD3在食管鳞状细胞癌（ESCC）中的作用尚不清楚。
通过蛋白质组学和转录组学分析确定ESCC与相邻正常组织之间的差异表达基因。然后对这些基因进行生存分析以确定预后标志物。评估两个亚组中的免疫细胞浸润情况。在TCGA-ESCC数据库中进行Spearman相关性分析，以检验PLOD3与RBM15表达之间的相关性。采用shRNA介导的方法敲低ESCC细胞中的RBM15。通过实时PCR和蛋白质印迹评估RBM15敲低对PLOD3表达的影响。此外，采用COX算法构建预后特征。
发现PLOD3在ESCC患者中高表达，并与良好预后相关。免疫细胞浸润估计表明，PLOD3高表达组中肿瘤浸润性CD4 + T细胞增加。相关性分析显示，PLOD3与RBM15相关，且与CD4 + T细胞浸润密切相关。此外，功能丧失方法表明，RBM15的缺失减弱了ESCC细胞中PLOD3的表达。经过单变量和多变量Cox回归分析，PLOD3和RBM15被确定为ESCC的双基因预后特征。
RBM15通过调节PLOD3增强ESCC中肿瘤浸润性CD4 + T细胞丰度。两个新的独立预后因素PLOD3和RBM15可能有助于预测ESCC的预后。
