Liu Yun, Li Lei, Wang Jia-Yao, Gao Fei, Lin Xia, Lin Shi-Shuai, Qiu Zhi-Yang, Liang Zun-Hong
Department of Plastic and Cosmetic Surgery, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, 570311, Hainan Province, People's Republic of China.
Department of Burn Plastic Surgery, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, No.19, Xiuhua Road, Xiuying District, Haikou, 570311, Hainan Province, People's Republic of China.
Mol Cell Biochem. 2023 Apr;478(4):707-719. doi: 10.1007/s11010-022-04538-6. Epub 2022 Aug 29.
Keloid is a common dermis tumor, occurring repeatedly, affecting the quality of patients' life. Long non-coding RNAs (lncRNAs) have crucial regulatory capacities in skin scarring formation and subsequent scar carcinogenesis. The intention of this study was to investigate the mechanism and function of GNAS antisense-1 (GNAS-AS1) in keloids. Clinical samples were collected to evaluate the expression of GNAS-AS1, RUNX2, and miR-188-5p by qRT-PCR. The proliferation, migration, and invasion of HKF cells were detected by CCK-8, wound healing, and Transwell assays. The expression levels of mRNA and protein were examined through qRT-PCR and Western blot assay. Luciferase reporter assay was used to identify the binding relationship among GNAS-AS1, miR-188-5p, and Runt-related transcription factor 2 (RUNX2). GNAS-AS1 and RUNX2 expressions were remarkably enhanced, and miR-188-5p expression was decreased in keloid clinical tissues and HKF cells. GNAS-AS1 overexpression promoted cells proliferation, migration, and invasion, while GNAS-AS1 knockdown had the opposite trend. Furthermore, overexpression of GNAS-AS1 reversed the inhibitory effect of 5-FU on cell proliferation, migration, and invasion. MiR-188-5p inhibition or RUNX2 overexpression could enhance the proliferation, migration, and invasion of HKF cells. GNAS-AS1 targeted miR-188-5p to regulate RUNX2 expression. In addition, the inhibition effects of GNAS-AS1 knockdown on HKF cells could be reversed by inhibition of miR-188-5p or overexpression of RUNX2, while RUNX2 overexpression eliminated the suppressive efficaciousness of miR-188-5p mimics on HKF cells growth. GNAS-AS1 knockdown could regulate the miR-188-5p/RUNX2 signaling axis to inhibit the growth and migration in keloid cells. It is suggested that GNAS-AS1 may become a new target for the prevention and treatment of keloid.
瘢痕疙瘩是一种常见的真皮肿瘤，易反复发生，影响患者生活质量。长链非编码RNA（lncRNA）在皮肤瘢痕形成及随后的瘢痕癌变过程中具有关键的调控能力。本研究旨在探究GNAS反义RNA-1（GNAS-AS1）在瘢痕疙瘩中的作用机制和功能。收集临床样本，采用qRT-PCR法评估GNAS-AS1、RUNX2和miR-188-5p的表达。通过CCK-8、伤口愈合试验和Transwell试验检测人正常皮肤成纤维细胞（HKF细胞）的增殖、迁移和侵袭能力。通过qRT-PCR和蛋白质免疫印迹法检测mRNA和蛋白质的表达水平。采用荧光素酶报告基因检测法确定GNAS-AS1、miR-188-5p和 runt相关转录因子2（RUNX2）之间的结合关系。在瘢痕疙瘩临床组织和HKF细胞中，GNAS-AS1和RUNX2的表达显著增强，而miR-188-5p的表达降低。过表达GNAS-AS1可促进细胞增殖、迁移和侵袭，而敲低GNAS-AS1则产生相反的作用。此外，过表达GNAS-AS1可逆转5-氟尿嘧啶对细胞增殖、迁移和侵袭的抑制作用。抑制miR-188-5p或过表达RUNX2可增强HKF细胞的增殖、迁移和侵袭能力。GNAS-AS1靶向miR-188-5p以调控RUNX2的表达。此外，抑制miR-188-5p或过表达RUNX2可逆转敲低GNAS-AS1对HKF细胞的抑制作用，而过表达RUNX2可消除miR-188-5p模拟物对HKF细胞生长的抑制作用。敲低GNAS-AS1可调节miR-188-5p/RUNX2信号轴，抑制瘢痕疙瘩细胞的生长和迁移。提示GNAS-AS1可能成为预防和治疗瘢痕疙瘩的新靶点。
