Zhang Qi, Feng Yingfa, Feng Jiangang, Zhang Jinming, Huang Lili
Department of Ultrasound, Hebei Chest Hospital, Shijiazhuang, Hebei, China.
Department of Orthopedics, The Fourth Hospital of Hebei Medical University, No. 12, Jiankang Road, Shijiazhuang, 050011 Hebei, China.
Open Life Sci. 2021 May 22;16(1):482-494. doi: 10.1515/biol-2021-0030. eCollection 2021.
Circular RNAs play crucial roles in tumor occurrence and progression. This research aimed to explore the role and potential mechanism of hsa_circ_0013359 (circ_0013359) in melanoma.
The levels of circ_0013359, microRNA-136-5p (miR-136-5p), and member RAS oncogene family (RAB9A) in melanoma tissues and cells were detected using quantitative reverse transcriptase-polymerase chain reaction or western blot. Cell proliferation, apoptosis, cell cycle, cell migration, and invasion were evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2--tetrazolium bromide assay, colony formation assay, flow cytometry, and transwell assay. Glycolysis was determined by detecting glucose consumption, lactate production, and extracellular acidification rate. The levels of hexokinase 2 and lactate dehydrogenase A were examined by western blot. The targeting relationship between miR-136-5p and circ_0013359 or RAB9A was confirmed by dual-luciferase reporter assay. Xenograft experiments were used to analyze tumor growth .
Circ_0013359 and RAB9A levels were increased, while the miR-136-5p level was reduced in melanoma tissues and cells. Circ_0013359 knockdown inhibited proliferation, migration, invasion, and glycolysis and promoted apoptosis and cycle arrest in A875 and SK-MEL-1 cells. Circ_0013359 sponged miR-136-5p to regulate melanoma progression. In addition, miR-136-5p suppressed melanoma progression by targeting RAB9A. Besides, circ_0013359 silencing inhibited tumor growth .
Depletion of circ_0013359 hindered melanoma progression by regulating miR-136-5p/RAB9A axis.
环状RNA在肿瘤的发生和发展中起关键作用。本研究旨在探讨hsa_circ_0013359(circ_0013359)在黑色素瘤中的作用及潜在机制。
采用定量逆转录-聚合酶链反应或蛋白质免疫印迹法检测黑色素瘤组织和细胞中circ_0013359、微小RNA-136-5p(miR-136-5p)和RAS癌基因家族成员(RAB9A)的水平。通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-溴化四氮唑蓝法、集落形成试验、流式细胞术和Transwell试验评估细胞增殖、凋亡、细胞周期、细胞迁移和侵袭。通过检测葡萄糖消耗、乳酸生成和细胞外酸化率来测定糖酵解。通过蛋白质免疫印迹法检测己糖激酶2和乳酸脱氢酶A的水平。通过双荧光素酶报告基因试验证实miR-136-5p与circ_0013359或RAB9A之间的靶向关系。采用异种移植实验分析肿瘤生长情况。
黑色素瘤组织和细胞中circ_0013359和RAB9A水平升高,而miR-136-5p水平降低。敲低circ_0013359可抑制A875和SK-MEL-1细胞的增殖、迁移、侵袭和糖酵解,并促进细胞凋亡和细胞周期阻滞。Circ_0013359通过吸附miR-136-5p来调节黑色素瘤进展。此外,miR-136-5p通过靶向RAB9A抑制黑色素瘤进展。此外,沉默circ_0013359可抑制肿瘤生长。
circ_0013359的缺失通过调节miR-136-5p/RAB9A轴阻碍黑色素瘤进展。