circ_0001821靶向miR-203调控皮肤鳞状细胞癌A431细胞的增殖、凋 亡、迁移和侵袭

目的：探讨circ_0001821通过靶向miR-203调控皮肤鳞状细胞癌（cutaneous squamous cell carcinoma，CSCC）A431细胞 增殖、凋亡、迁移及侵袭的分子机制。方法：选取2018年2月至2019年8月海南医学院第二附属医院收治的39例CSCC患者的癌 及配对癌旁组织，以及人CSCC细胞系A431，用qPCR法检测CSCC癌和癌旁组织中circ_0001821的表达水平。利用脂质体转染技 术，分别将si-circ_0001821、si-NC、miR-203 mimic、miR-NC、si-circ_0001821与anti-miR-NC、si-circ_0001821与anti-miR-203转染至 A431细胞，用qPCR法检测转染细胞中circ_0001821和miR-203的表达水平，MTT法、流式细胞术和Transwell实验分别检测A431 细胞的增殖、凋亡、迁移及侵袭能力，WB法检测细胞中增殖、凋亡、迁移及侵袭相关蛋白的表达水平。通过Circinteractome数据库 预测circ_0001821与miR-203存在结合位点，双荧光素酶报告基因实验验证circ_0001821与miR-203的靶向关系。结果：CSCC组 织中circ_0001821的表达水平显著高于癌旁组织（P<0.01）。转染 si-circ_0001821 可显著降低细胞中circ_0001821的表达水平（P<0.01），降低细胞增殖、迁移和侵袭能力（均P<0.01），提高细胞凋亡率（P<0.01）。双荧光素酶报告基因实验证实，circ_0001821可 靶向结合miR-203。转染miR-203 mimic可显著降低A431细胞的增殖、迁移和侵袭能力（均P<0.01），提高细胞凋亡率（P<0.01）。共转染si-circ_0001821与anti-miR-203 可明显逆转下调circ_0001821 表达对 A431 细胞增殖、迁移、侵袭及凋亡的调控作用。结论： circ_0001821通过靶向miR-203调控CSCC细胞A431的增殖、迁移、侵袭能力及细胞凋亡。

circ_0001821 regulates the proliferation, apoptosis, migration and invasion of cutaneous squamous cell carcinoma A431 cells by targeting miR-203

Objective: To explore the mechanism of circ_0001821 regulating the proliferation, apoptosis, migration and invasion of cutaneous squamous cell carcinoma (CSCC) A431 cells by targeting miR-203. Methods: The cancer and para-cancerous tissues of 39 patients with CSCC treated in the Second Affiliated Hospital of Hainan Medical University from February 2018 to August 2019, as well as the human CSCC cell line A431, were collected for this study. The qPCR method was used to detect the expression level of circ_0001821 in CSCC tissues and para-cancerous tissues. Using liposome transfection technology, si-circ_0001821, si-NC, miR-203 mimic, miR-NC, si-circ_0001821+anti-miR-NC and si-circ_0001821+anti-miR-203 were transfected into A431 cells, respectively. qPCR was used to detect the expression of circ_0001821 and miR-203 in the transfected cells. MTT method, Flow cytometry and Transwell chamber assay were used to detect the proliferation, apoptosis, migration and invasion of transfected A431 cells. WB was used to detect the expression of proteins related to proliferation, apoptosis, migration and invasion. Circinteractome database was utilized to predict the binding site between circ_0001821 and miR-203, which was further validated by Dual-luciferase reporter gene assay. Results: Compared with para-cancerous tissues, the expression level of circ_0001821 in CSCC tissues was significantly increased (P<0.01). Transfection of si-circ_0001821 could significantly reduce the expression of circ_0001821 (P<0.01), increase the apoptosis rate (P<0.01), and reduce the proliferation, migration and invasion abilities of A431 cells (all P<0.01). Dual-luciferase report gene assay confirmed that circ_0001821 could targetedly bind with miR-203. Transfection of miR-203 mimics could significantly reduce the proliferation, migration and invasion abilities but increase the apoptosis rate of A431 cells (all P<0.01). . Co-transfection of si-circ_0001821 and anti-miR-203 could significantly reverse the effects of si-circ_0001821 transfection on the proliferation, migration, invasion and apoptosis of A431 cells. Conclusion: circ_0001821 regulates the proliferation, migration, invasion and apoptosis of CSCC A431 cells by targeting miR-203.