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Abstract：
OBJECTIVE: LncRNA HCG18 has been reported to act as a tumor promoter in gastric cancer, hepatocellular carcinoma and nasopharyngeal carcinoma. However, the role of HCG18 in melanoma is still not clear. In this study, we detected the expression and molecular function of HCG18 in melanoma.
METHODS: The expression of HCG18 in melanoma cell lines and 50 pairs of melanoma and corresponding non-cancer tissues was detected by RT-qPCR. The relationship between HCG18 and clinicopathology was analyzed. We used HCG18 overexpressing melanoma cell lines A375 and M14, and si-HCG18 to knock down HCG18 expression. CCK-8, clone formation, Transwell assay and FCM were used to explore the effect of HCG18 knockdown on cell proliferation, migration, invasion and apoptosis in melanoma cells. Bioinformatics software was used to predict the downstream miRNA regulated by HCG18, and the downstream target genes regulated by miR-324-5p. Dual-luciferase reporter assay and RNA pull-down assay were used to verify whether miR-324-5p was related to the predicted sequence of HCG18.
RESULTS: HCG18 was highly expressed in melanoma tissues and cells. Besides, we found that HCG18 was closely correlated with thickness, TNM stage and metastasis. Functional experiments discovered that HCG18 knockdown restrained cell proliferation, migration and invasion, while promoted cell apoptosis in melanoma cells. HCG18 was confirmed to be a sponge of miR-324-5p, and CDK16 might be a downstream gene of miR-324-5p. HCG18 was found to reverse the effect of miR-324-5p by upregulating CDK16 expression in melanoma cell proliferation, apoptosis, migration and invasion in vitro.
CONCLUSIONS: This study indicated that HCG18 played an essential role in the pathogenesis of melanoma and suggested that HCG18 might be a potential target for the treatment and diagnosis of melanoma.
METHODS: The expression of HCG18 in melanoma cell lines and 50 pairs of melanoma and corresponding non-cancer tissues was detected by RT-qPCR. The relationship between HCG18 and clinicopathology was analyzed. We used HCG18 overexpressing melanoma cell lines A375 and M14, and si-HCG18 to knock down HCG18 expression. CCK-8, clone formation, Transwell assay and FCM were used to explore the effect of HCG18 knockdown on cell proliferation, migration, invasion and apoptosis in melanoma cells. Bioinformatics software was used to predict the downstream miRNA regulated by HCG18, and the downstream target genes regulated by miR-324-5p. Dual-luciferase reporter assay and RNA pull-down assay were used to verify whether miR-324-5p was related to the predicted sequence of HCG18.
RESULTS: HCG18 was highly expressed in melanoma tissues and cells. Besides, we found that HCG18 was closely correlated with thickness, TNM stage and metastasis. Functional experiments discovered that HCG18 knockdown restrained cell proliferation, migration and invasion, while promoted cell apoptosis in melanoma cells. HCG18 was confirmed to be a sponge of miR-324-5p, and CDK16 might be a downstream gene of miR-324-5p. HCG18 was found to reverse the effect of miR-324-5p by upregulating CDK16 expression in melanoma cell proliferation, apoptosis, migration and invasion in vitro.
CONCLUSIONS: This study indicated that HCG18 played an essential role in the pathogenesis of melanoma and suggested that HCG18 might be a potential target for the treatment and diagnosis of melanoma.
摘要：
目的：据报道，LncRNAHCG18可作为胃癌的肿瘤启动子，肝细胞癌和鼻咽癌。然而,HCG18在黑色素瘤中的作用尚不清楚。在这项研究中,我们检测了HCG18在黑色素瘤中的表达和分子功能。
方法：通过RT-qPCR检测HCG18在黑色素瘤细胞系和50对黑色素瘤及相应的非癌组织中的表达。分析HCG18与临床病理的关系。我们使用过表达HCG18的黑素瘤细胞系A375和M14以及si-HCG18来敲低HCG18表达。CCK-8，克隆形成，用Transwell法和FCM法探讨HCG18基因敲低对细胞增殖的影响,迁移,黑色素瘤细胞的侵袭和凋亡。应用生物信息学软件预测HCG18调控的下游miRNA和miR-324-5p调控的下游靶基因。使用双荧光素酶报告基因测定和RNA下拉测定来验证miR-324-5p是否与HCG18的预测序列相关。
结果：HCG18在黑色素瘤组织和细胞中高表达。此外,我们发现HCG18与厚度密切相关，TNM分期和转移。功能实验发现HCG18敲低抑制细胞增殖,移民和入侵，同时促进黑色素瘤细胞凋亡。HCG18被证实是miR-324-5p的海绵，CDK16可能是miR-324-5p的下游基因。HCG18被发现通过上调黑色素瘤细胞增殖中的CDK16表达来逆转miR-324-5p的作用,凋亡，体外迁移和侵袭。
结论：这项研究表明HCG18在黑色素瘤的发病机制中起着至关重要的作用，提示HCG18可能是黑色素瘤治疗和诊断的潜在靶标。
方法：通过RT-qPCR检测HCG18在黑色素瘤细胞系和50对黑色素瘤及相应的非癌组织中的表达。分析HCG18与临床病理的关系。我们使用过表达HCG18的黑素瘤细胞系A375和M14以及si-HCG18来敲低HCG18表达。CCK-8，克隆形成，用Transwell法和FCM法探讨HCG18基因敲低对细胞增殖的影响,迁移,黑色素瘤细胞的侵袭和凋亡。应用生物信息学软件预测HCG18调控的下游miRNA和miR-324-5p调控的下游靶基因。使用双荧光素酶报告基因测定和RNA下拉测定来验证miR-324-5p是否与HCG18的预测序列相关。
结果：HCG18在黑色素瘤组织和细胞中高表达。此外,我们发现HCG18与厚度密切相关，TNM分期和转移。功能实验发现HCG18敲低抑制细胞增殖,移民和入侵，同时促进黑色素瘤细胞凋亡。HCG18被证实是miR-324-5p的海绵，CDK16可能是miR-324-5p的下游基因。HCG18被发现通过上调黑色素瘤细胞增殖中的CDK16表达来逆转miR-324-5p的作用,凋亡，体外迁移和侵袭。
结论：这项研究表明HCG18在黑色素瘤的发病机制中起着至关重要的作用，提示HCG18可能是黑色素瘤治疗和诊断的潜在靶标。
