Tsai Kuo-Wang, Liao Jia-Bin, Tseng Hui-Wen
Department of Research, Taipei Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, New Taipei City, Taiwan.
Department of Nursing, Cardinal Tien Junior College of Healthcare and Management, New Taipei City, Taiwan.
Cancer Cell Int. 2024 Nov 19;24(1):384. doi: 10.1186/s12935-024-03545-5.
Melanoma is an aggressive tumor with a high mortality rate. Metformin, a commonly prescribed diabetes medication, has shown promise in cancer prevention and treatment. Long noncoding RNAs (lncRNAs) are non-protein-coding RNA molecules that play a key role in tumor development by interacting with cellular chromatins. Despite the benefits of metformin, the anticancer mechanism underlying its effect on the regulation of lncRNAs in melanoma remains unclear.
We investigated the lncRNA profiles of human melanoma cells with and without metformin treatment using a next-generation sequencing approach (NGS). Utilizing public databases, we analyzed the expression levels and clinical impacts of LINC00094 and miR-1270 in melanoma. The expression levels of LINC00094 and miR-1270 were verified in human cell lines and clinical samples by real-time PCR and in situ hybridization. The biological roles of LINC00094 and miR-1270 in cell growth, proliferation, cell cycle, apoptosis, and motility were studied using in vitro assays.
We identify a novel long noncoding RNA, namely LINC00094, whose expression considerably decreased in melanoma cells after metformin treatment. In situ hybridization analysis revealed substantially higher expression of LINC00094 in cutaneous melanoma tissue compared with adjacent normal epidermis and normal control tissues (P < 0.001). In nondiabetic patients with melanoma, the overall survival of high LINC00094 expression group was shorter than the low LINC00094 expression group with borderline statistical significance (log-rank test, P = 0.057). Coexpression analysis of LINC00094 indicated its involvement in the mitochondrial respiratory pathway, with its knockdown suppressing genes associated with mitochondrial oxidative phosphorylation, glycolysis, antioxidant production, and metabolite levels. Functional analysis revealed that silencing-LINC00094 inhibited the proliferation, colony formation, invasion, and migration of melanoma cells. Cell cycle analysis following LINC00094 knockdown revealed G1 phase arrest with reduced cell cycle protein expression. Combined TargetScan and reporter assays revealed a direct link between miR-1270 and LINC00094. Ectopic miR-1270 expression inhibited melanoma cell growth and motility while inducing apoptosis. Finally, through in silico analysis, we identified two miR-1270 target genes, CD276 and centromere protein M (CENPM), which may be involved in the biological functions of LINC00094.
Overall, LINC00094 expression may regulate melanoma cell growth and motility by modulating the expression of miR-1270, and targeting genes of CD276 and CENPM indicating its therapeutic potential in melanoma treatment.
黑色素瘤是一种侵袭性肿瘤，死亡率很高。二甲双胍是一种常用的糖尿病药物，在癌症预防和治疗方面显示出前景。长链非编码RNA（lncRNA）是一类非蛋白质编码的RNA分子，通过与细胞染色质相互作用在肿瘤发展中起关键作用。尽管二甲双胍有诸多益处，但其对黑色素瘤中lncRNA调控的抗癌机制仍不清楚。
我们采用下一代测序方法（NGS）研究了经二甲双胍处理和未经处理的人黑色素瘤细胞的lncRNA谱。利用公共数据库，我们分析了LINC00094和miR - 1270在黑色素瘤中的表达水平及临床影响。通过实时PCR和原位杂交在人细胞系和临床样本中验证了LINC00094和miR - 1270的表达水平。使用体外实验研究了LINC00094和miR - 1270在细胞生长、增殖、细胞周期、凋亡和迁移中的生物学作用。
我们鉴定出一种新型长链非编码RNA，即LINC00094，其在二甲双胍处理后的黑色素瘤细胞中表达显著降低。原位杂交分析显示，与相邻正常表皮和正常对照组织相比，皮肤黑色素瘤组织中LINC00094的表达明显更高（P < 0.001）。在非糖尿病黑色素瘤患者中，高LINC00094表达组的总生存期短于低LINC00094表达组，具有临界统计学意义（对数秩检验，P = 0.057）。LINC00094的共表达分析表明其参与线粒体呼吸途径，其敲低可抑制与线粒体氧化磷酸化、糖酵解、抗氧化剂产生和代谢物水平相关的基因。功能分析表明，沉默LINC00094可抑制黑色素瘤细胞的增殖、集落形成、侵袭和迁移。LINC00094敲低后的细胞周期分析显示G1期阻滞，细胞周期蛋白表达降低。联合TargetScan和报告基因分析揭示了miR - 1270与LINC00094之间的直接联系。异位表达miR - 1270可抑制黑色素瘤细胞生长和迁移，同时诱导凋亡。最后，通过计算机分析，我们鉴定出两个miR - 1270靶基因，CD276和着丝粒蛋白M（CENPM），它们可能参与LINC00094的生物学功能。
总体而言，LINC00094的表达可能通过调节miR - 1270的表达来调控黑色素瘤细胞的生长和迁移，并且靶向CD276和CENPM基因表明其在黑色素瘤治疗中的潜在治疗价值。
