Xie Jing, Zheng Yanyan, Xu Xiaomin, Sun Congcong, Lv Mingfen
Department of Dermatology, The Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou People's Hospital, Wenzhou 325000, People's Republic of China.
Department of Neurology, The Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou People's Hospital, Wenzhou 325000, People's Republic of China.
Onco Targets Ther. 2020 Jun 29;13:6203-6211. doi: 10.2147/OTT.S249736. eCollection 2020.
Melanoma is a very malignant skin cancer with high mortality and unsatisfactory prognosis. Many long noncoding RNAs (lncRNAs) have been reported to be aberrantly expressed in melanoma. How lncRNA regulates melanoma progression is poorly defined. LncRNA CAR10 has been shown to regulate the progression of several cancers and its role in melanoma remains unclear. This study aims to determine the role and mechanism of lncRNA CAR10 in the regulation of melanoma progression.
qRT-PCR was utilized to analyze CAR10 in melanoma human tissues and cell lines while Kaplan-Meier curve was used to examine the survival rate. CCK8 assay and EdU assay were used to assess cell proliferation when Transwell assay was conducted to determine migration and invasion. And tumor xenograft assay was performed to evaluate tumor growth in vivo. Additionally, luciferase assay and RNA pulldown assay were performed to analyze the interactions among CAR10, miR-125b-5p and RAB3D.
LncRNA CAR10 was upregulated in melanoma tissues and cell lines. Upregulation of CAR10 predicted a poor prognosis in patients with melanoma. CAR10 knockdown suppressed proliferation, migration and invasion of melanoma cells in vitro. CAR10 silencing attenuated tumor growth in vivo. CAR10 inhibited miR-125b-5p activity to upregulate RAB3D expression. And miR-125b-5p/RAB3D signaling is crucial for CAR10-dependent melanoma progression.
Our work suggests that lncRNA CAR10 promotes melanoma growth and metastasis through modulating miR-125b-5p/RAB3D axis.
黑色素瘤是一种恶性程度很高的皮肤癌，死亡率高且预后不理想。许多长链非编码RNA（lncRNA）已被报道在黑色素瘤中异常表达。lncRNA如何调节黑色素瘤的进展尚不清楚。lncRNA CAR10已被证明可调节多种癌症的进展，其在黑色素瘤中的作用仍不清楚。本研究旨在确定lncRNA CAR10在调节黑色素瘤进展中的作用和机制。
采用qRT-PCR分析黑色素瘤人体组织和细胞系中的CAR10，同时用Kaplan-Meier曲线检测生存率。用CCK8法和EdU法评估细胞增殖，用Transwell法检测迁移和侵袭。并进行肿瘤异种移植试验以评估体内肿瘤生长。此外，进行荧光素酶试验和RNA下拉试验以分析CAR10、miR-125b-5p和RAB3D之间的相互作用。
lncRNA CAR10在黑色素瘤组织和细胞系中上调。CAR10的上调预示着黑色素瘤患者预后不良。敲低CAR10可抑制黑色素瘤细胞在体外的增殖、迁移和侵袭。沉默CAR10可减弱体内肿瘤生长。CAR10抑制miR-125b-5p活性以上调RAB3D表达。并且miR-125b-5p/RAB3D信号通路对CAR10依赖的黑色素瘤进展至关重要。
我们的工作表明，lncRNA CAR10通过调节miR-125b-5p/RAB3D轴促进黑色素瘤的生长和转移。
