The role and molecular mechanism of CD73 in melanoma
10.3760/cma.j.cn114453-20210530-00235
- VernacularTitle:CD73在黑色素瘤中的作用及分子机制研究
- Author:
Zixu GAO
1
;
Lu WANG
;
Xinyi DENG
;
Ming REN
;
Chuanyuan WEI
;
Jianying GU
Author Information
1. 复旦大学附属中山医院整形外科，上海 200032
- Keywords:
Melanoma;
Cell proliferation;
Neoplasm invasiveness;
CD73
- From:
Chinese Journal of Plastic Surgery
2021;37(8):884-893
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the expression level of CD73 in cutaneous melanoma and its clinical significance, and to explore its role and molecular mechanism in the occurrence and development of melanoma.Methods:Expression level of the gene NT5E in melanoma was analyzed according to RNA sequencing data from the CCLE database. CD73 expression level was tested in 145 melanoma tissues and corresponding para-tumor tissues by immunohistochemistry tissue microarray. The samples came from 78 male and 67 female patients who received surgical resection and were pathologically diagnosed as melanoma in the department of plastic surgery in Zhongshan Hospital from January of 2008 to December of 2017. The relationship between CD73 expression level and clinicopathological features as well as prognosis was analyzed. Melanoma cell line was transfected with CD73 gene overexpression lentivirus and negative control vectors, respectively. The transfection efficiency was verified by qRT-PCR and Western blot. CCK-8, wound healing and Transwell assays were used to evaluate the proliferation, migration and invasion abilities of the overexpression group and control group. Subcutaneous xenograft tumor models were established in nude mice to assess the tumorigenicity of the two groups. Transcriptome sequencing, GO and KEGG analyses were performed. The expression levels of pAKT and pGSK3β were detected by Western blot. The measurement data were expressed as Mean ±SD. Independent sample t-test was used for comparison between the two groups, and One-way ANOVA was employed for multi-group comparisons. The chi-square test was used for the categorical variables. P<0.05 was considered as statistically significant difference. Results:The expression of CD73 in melanoma was higher than that in para-tumor tissues ( P< 0.05). Higher expression level of CD73 was associated with higher Clark grade ( P= 0.014) and clinical stage ( P= 0.040). The overall survival and disease-free survival of patients with high-CD73-expression were significantly lower than those of patients with low-CD73-expression. CD73 expression, Clark grade and clinical stage are independent risk factors for the poor prognosis of patients with melanoma. Overexpression of CD73 significantly enhanced the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo( P< 0.05). Transcriptome sequencing showed that CD73 was involved in several tumor related signaling pathways, among which the expression of pAKT and pGSK3β was elevated( P< 0.05). Conclusions:CD73 promotes the proliferation, migration and invasion of melanoma cells in vitro and the tumorigenicity in vivo through activating AKT signaling. CD73 is expected to become a new prognostic biomarker and therapeutic target for melanoma.
