Li B, Zhou C X, Pu Y Q, Qiu L, Mei W, Xiong W
College of Basic Medical Sciences, Dali University, Dali 671000, China Key Laboratory of Clinical Biochemical Testing in Yunnan Province Universities, Dali 671000, China.
College of Chemistry and Life Sciences, Chuxiong Normal University, Chuxiong 675000, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2023 Mar 20;41(3):168-176. doi: 10.3760/cma.j.cn121094-20220228-00100.
To investigate the expression of CD24 gene in human malignant pleural mesothelioma (MPM) cells and tissues, and evaluate its relationship with clinicopathological characteristics and clinical prognosis of MPM patients. In February 2021, UALCAN database was used to analyze the correlation between CD24 gene expression and clinicopathological characteristics in 87 cases of MPM patients. The TIMER 2.0 platform was used to explore the relationship between the expression of CD24 in MPM and tumor immune infiltrating cells. cBioportal online tool was used to analyze the correlation between CD24 and MPM tumor marker gene expression. RT-qPCR was used to analyze the expressions of CD24 gene in human normal pleural mesothelial cell lines LP9 and MPM cell lines NCI-H28 (epithelial type), NCI-H2052 (sarcoma type), and NCI-H2452 (biphasic mixed type). RT-qPCR was performed to detect the expressions of CD24 gene in 18 cases of MPM tissues and matched normal pleural tissues. The expression difference of CD24 protein in normal mesothelial tissue and MPM tissue was analyzed by immunohistochemistry. A Kaplan-Meier model was constructed to explore the influence of CD24 gene expression on the prognosis of MPM patients, and Cox regression analysis of prognostic factors in MPM patients was performed. The CD24 gene expression without TP53 mutation MPM patients was significantly higher than that of patients in TP53 mutation (<0.05). The expression of CD24 gene in MPM was positively correlated with B cells ((s)=0.37, <0.001). The expression of CD24 gene had a positive correlation with the expressions of thrombospondin 2 (THBS2) ((s)=0.26, <0.05), and had a negative correlation with the expression of epidermal growth factor containing fibulin like extracellular matrix protein 1 (EFEMP1), mesothelin (MSLN) and calbindin 2 (CALB2) ((s)=-0.31, -0.52, -0.43, <0.05). RT-qPCR showed that the expression level of CD24 gene in MPM cells (NCI-H28, NCI-H2052 and NCI-H2452) was significantly higher than that in normal pleural mesothelial LP9 cells. The expression level of CD24 gene in MPM tissues was significantly higher than that in matched normal pleural tissues (<0.05). Immunohistochemistry showed that the expressions of CD24 protein in epithelial and sarcoma MPM tissues were higher than those of matched normal pleural tissues. Compared with low expression of CD24 gene, MPM patients with high expression of CD24 gene had lower overall survival (=2.100, 95%: 1.336-3.424, <0.05) and disease-free survival (=1.800, 95%: 1.026-2.625, <0.05). Cox multivariate analysis showed that compared with the biphasic mixed type, the epithelial type was a protective factor for the prognosis of MPM patients (=0.321, 95%: 0.172-0.623, <0.001). Compared with low expression of CD24 gene, high expression of CD24 gene was an independent risk factor for the prognosis of MPM patients (=2.412, 95%: 1.291-4.492, =0.006) . CD24 gene and protein are highly expressed in MPM tissues, and the high expression of CD24 gene suggests poor prognosis in MPM patients.
探讨CD24基因在人恶性胸膜间皮瘤（MPM）细胞和组织中的表达，并评估其与MPM患者临床病理特征及临床预后的关系。2021年2月，利用UALCAN数据库分析87例MPM患者CD24基因表达与临床病理特征之间的相关性。使用TIMER 2.0平台探讨MPM中CD24表达与肿瘤免疫浸润细胞之间的关系。利用cBioportal在线工具分析CD24与MPM肿瘤标志物基因表达之间的相关性。采用RT-qPCR分析人正常胸膜间皮细胞系LP9及MPM细胞系NCI-H28（上皮型）、NCI-H2052（肉瘤型）和NCI-H2452（双相混合型）中CD24基因的表达。采用RT-qPCR检测18例MPM组织及配对正常胸膜组织中CD24基因的表达。通过免疫组化分析正常间皮组织和MPM组织中CD24蛋白的表达差异。构建Kaplan-Meier模型探讨CD24基因表达对MPM患者预后的影响，并对MPM患者的预后因素进行Cox回归分析。无TP53突变的MPM患者CD24基因表达显著高于TP53突变患者（<0.05）。MPM中CD24基因的表达与B细胞呈正相关（(s)=0.37，<0.001）。CD24基因的表达与血小板反应蛋白2（THBS2）的表达呈正相关（(s)=0.26，<0.05），与含表皮生长因子的纤连蛋白样细胞外基质蛋白1（EFEMP1）、间皮素（MSLN）和钙结合蛋白2（CALB2）的表达呈负相关（(s)=-0.31、-0.52、-0.43，<0.05）。RT-qPCR显示，MPM细胞（NCI-H28、NCI-H2052和NCI-H2452）中CD24基因的表达水平显著高于正常胸膜间皮LP9细胞。MPM组织中CD24基因的表达水平显著高于配对的正常胸膜组织（<0.05）。免疫组化显示，上皮型和肉瘤型MPM组织中CD24蛋白的表达高于配对的正常胸膜组织。与CD24基因低表达的MPM患者相比，CD24基因高表达的MPM患者总生存期较低（=2.100，95%：1.336 - 3.424，<0.05），无病生存期较低（=1.800，95%：1.026 - 2.625，<0.05）。Cox多因素分析显示，与双相混合型相比，上皮型是MPM患者预后的保护因素（=0.321，95%：0.172 - 0.623，<0.001）。与CD24基因低表达相比，CD24基因高表达是MPM患者预后的独立危险因素（=2.412，95%：1.291 - 4.492，=0.006）。CD24基因和蛋白在MPM组织中高表达，CD24基因高表达提示MPM患者预后不良。
