CCL2在骨肉瘤组织中的表达及其临床意义

Clinical significance of CCL2 expression in osteosarcoma tissue

背景与目的:骨肉瘤是一种原发于骨组织的恶性肿瘤,主要发病于青少年,发生肺转移的患者预后较差。CC族趋化因子配体2(CC chemokine ligand 2,CCL2)及其受体CC族趋化因子受体2(CC chemokine ligand receptor 2,CCR2),与肿瘤的发生、发展密切相关。已经有学者证实,CCL2会促进乳腺癌的转移。探讨CCL2在骨肉瘤组织中的表达及其与预后的关系。方法:应用GEXSCOPE®组织解离液将11例骨肉瘤组织解离成单细胞悬液,应用美国10x Genomics公司试剂盒(Chromium Single Cell Gene Expression Kit、Gel Bead Kit及Multiplex Kit)建立单细胞RNA测序(single cell RNA sequence,scRNA-seq)文库,分析样本CCL2表达。收集2017年1月—2019年12月在上海交通大学附属第六人民医院治疗的骨肉瘤患者90例的石蜡组织,制备组织芯片,采用免疫组织化学方法检测CLL2的表达并评分,以所有结果的平均分为高表达和低表达的划分值,采用COX回归分析其与临床病理学特征及预后的关系。结果:scRNA-seq单细胞检测结果显示,与肿瘤原发灶相比,骨肉瘤肺转移的患者CCL2表达更高。组织芯片分析可评价的80例患者中CCL2高表达患者33例,低表达47例,CCL2的表达与临床分期和远处转移相关(P<0.05),与年龄、性别、肿瘤大小、病理学类型及肿瘤坏死率无明显相关性(P>0.05)。结论:CCL2可能是判断骨肉瘤转移及预后的指标之一。

Background and purpose:Osteosarcoma is a primary malignant tumor in bone tissue,mainly in adolescents.The prognosis of patients with lung metastasis is poor.CC chemokine ligand 2(CCL2)and tis receptor CC chemokine ligand receptor 2(CCR2)may take part in tumor.Some scholars have confirmed that CCL2 can promote the metastasis process of breast cancer and worsen the prognosis.The main purpose of this article was to study the expression of CCL2 in osteosarcoma tissue and its relationship with prognosis.Methods:GEXSCOPE®tissue dissociation solution was used to dissociate 11 osteosarcoma samples into single cell suspension.The single cell sequencing library(scRNA-seq)was established by Chromium Single Cell Gene Expression Kit,Gel Bead Kit and Multiplex Kit from 10x Genomics Company.The expression of CCL2 was analyzed by scRNA-seq.The CCL2 level in lesions from tissue microarray was detected in 90 osteosarcoma patients,who were diagnosed and treated in the 6th People’s Hospital Affiliated to Shanghai Jiao Tong University from January 2017 to December 2019,using immunohistochemical method.According to the median value of the CCL2 histochemical score,90 patients were divided into two groups,high expression and low expression.COX regression was used to analyze the CCL2 level and its relationship with clinicopathological characteristics and prognosis of patient.Results:The results of scRNA-seq indicated that CCL2 expression was higher in lung metastasis lesions than in primary tissues.The immunohistochemical results displayed that of the 80 cases,33 had high CCL2 expression level and 47 had low CCL2 expression level.CCL2 expression was associated with clinical stage and lung metastasis(P<0.05).There was no significant correlation among the CCL2 level and age,gender,tumor size,pathological type and tumor necrosis rate(P>0.05).Conclusion:CCL2 may be one of the poor indicators to predict the metastasis and prognosis of osteosarcoma.