LINC01296 通过调控 miR-324-3p/MAPK1 轴促进皮肤恶性黑色素瘤的增殖。

LINC01296 promotes proliferation of cutaneous malignant melanoma by regulating miR-324-3p/MAPK1 axis.

机构信息

Department of General Surgery, Gaoyou People’s Hospital, Yangzhou 225600, China.

Department of Dermatology, The First People’s Hospital of Lianyungang, Lianyungang Clinical Medical College of Nanjing Medical University, Lianyungang 222002, China.

出版信息

Aging (Albany NY). 2022 Dec 2;15(8):2877-2890. doi: 10.18632/aging.204413.

DOI:10.18632/aging.204413

PMID:36462499

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10188354/

Abstract

OBJECTIVE

To investigate the functions and potential molecular mechanism of LINC01296 regarding the progression of cutaneous malignant melanoma (CMM) by the regulation of miR-324-3p/MAPK1 axis.

METHODS

The candidate differential lncRNAs of CMM were selected from GEPIA database, and quantitative real-time PCR (qRT-PCR) was utilized to assess the expression level of LINC01296 in human CMM tissues and cell lines. Cell proliferation assay, Colony formation assay, Ethynyl-2'-deoxyuridine (EDU) assay and tumorigenicity assays in nude mice were performed to examine the functions of LINC01296. Bioinformatics analysis, luciferase reporter assay and rescue experiments were also gained an insight into the underlying mechanisms of LINC01296 in CMM cell lines by miR-324-3p/MAPK1 axis.

RESULTS

In this study, the up-regulation of LINC01296 was found in CMM tissues and cell lines. Functionally, the over-expression of LINC01296 promoted the proliferation in CMM cell lines. In addition, immunochemistry analysis confirmed that the levels of MAPK1 and Ki-67 in sh-LINC01296-xenografted tumors was weaker than that in sh-NC-xenografted tumors. Then, bioinformatics analysis confirmed that LINC01296 interacted with miR-324-3p. Further investigations showed that MAPK1, which collected from the potential related genes of LINC01296, was the conjugated mRNA of miR-324-3p by luciferase reporter assay. Finally, the rescue experiments suggested the positive regulatory association among LINC01296 and MAPK1, which showed that MAPK1 could reverse the promoting-effect of LINC01296 in CMM cells .

CONCLUSIONS

Therefore, our findings provided insight into the mechanisms of LINC01296 via miR-324-3p/MAPK1 axis in CMM, and revealed an alternative target for the diagnosis and treatment of CMM.

摘要

目的

通过调节 miR-324-3p/MAPK1 轴，研究 LINC01296 在皮肤恶性黑色素瘤（CMM）进展中的功能和潜在分子机制。

方法

从 GEPIA 数据库中选择候选差异 lncRNA，并用实时定量 PCR（qRT-PCR）评估 LINC01296 在人 CMM 组织和细胞系中的表达水平。细胞增殖实验、集落形成实验、Ethynyl-2'-deoxyuridine (EDU) 实验和裸鼠肿瘤生成实验用于研究 LINC01296 的功能。通过 miR-324-3p/MAPK1 轴进行生物信息学分析、荧光素酶报告实验和挽救实验，探讨 LINC01296 在 CMM 细胞系中的潜在机制。

结果

本研究发现 LINC01296 在 CMM 组织和细胞系中上调。功能上，LINC01296 的过表达促进了 CMM 细胞系的增殖。此外，免疫组织化学分析证实，sh-LINC01296-异种移植瘤中 MAPK1 和 Ki-67 的水平弱于 sh-NC-异种移植瘤。然后，生物信息学分析证实 LINC01296 与 miR-324-3p 相互作用。进一步的研究表明，MAPK1 是从 LINC01296 的潜在相关基因中收集到的，通过荧光素酶报告实验证实其是 miR-324-3p 的结合 mRNA。最后，挽救实验表明 LINC01296 和 MAPK1 之间存在正调控关联，表明 MAPK1 可以逆转 LINC01296 在 CMM 细胞中的促进作用。