Long Non-coding RNA LINC-PINT Suppresses Cell Proliferation and Migration of Melanoma via Recruiting EZH2.

Yangfan Xu,Fang Li,Huixue Wang,Renbing Jia,Jie Yang,Xiaoyu He,Shengfang Ge,Jing Ruan,Ludwig M Heindl,Xianqun Fan,Jie Yu

doi:10.3389/fcell.2019.00350

Yangfan Xu, Fang Li + Show 9 more

Open Access

https://doi.org/10.3389/fcell.2019.00350

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Abstract

Long non-coding RNAs (lncRNAs) have been identified as crucial regulators in many human cancers. Many lncRNAs show aberrant expression in cancer, and some of them play critical roles in tumor proliferation, invasion, and metastasis. However, the regulatory functions of lncRNAs in melanoma progression remain to be elucidated. We utilized the Real-time PCR methodology to determine the expression of LINC-PINT in melanoma cell lines. To evaluate the effect of LINC-PINT on tumorigenesis of melanoma, we used Cell Counting Kit-8 (CCK8) and colony formation assay. Flow cytometry assay was used to detect the function of LINC-PINT on cell cycle status. PINT-interacting proteins were identified by chromatin isolation using RNA purification (ChIRP). Microarray assay and bioinformatics analysis were used to find the potential target genes of LINC-PINT and the status of LINC-PINT target gene candidate was verified using chromatin immunoprecipitation assay (ChIP). LINC-PINT plays a role in suppressing the tumorigenicity of melanoma, which was further determined by xenograft model assay. LINC-PINT was significantly downregulated in melanoma tissues and cell lines. The overexpression of LINC-PINT in tumor cells resulted in significant tumor growth reduction and migration inhibition in A375, Mum2B and CRMM1 cells. Results based on the in vivo xenograft model were further consistent with the in vitro findings that LINC-PINT impeded growth and metastasis of melanoma cells. Microarray assay and bioinformatics analysis indicated that CDK1, CCNA2, AURKA, and PCNA were potential targets of LINC-PINT. In conclusion, LINC-PINT inhibits the tumorigenicity of melanoma through recruiting EZH2 to the promoter of its target genes, leading to H3K27 trimethylation and epigenetic silencing of target genes. LINC-PINT may serve as a novel diagnostic and therapeutic target for melanoma.

Highlights

Melanoma is a malignant tumor that initiates from pigment-producing cells called melanocytes and progresses in a step-wise fashion (Bastian, 2014)
We found that LINCPINT was one of the most downregulated Long non-coding RNAs (lncRNAs) in melanoma cells (Figure 1A)
LINC-PINT expression level was significantly low in melanoma tissues compared with adjacent normal tissues (Figure 1B)

Summary

Introduction

Melanoma is a malignant tumor that initiates from pigment-producing cells called melanocytes and progresses in a step-wise fashion (Bastian, 2014). Melanoma is exceedingly aggressive, which is based on. The Tumor-Suppressive Role of LINC-PINT in Melanoma the high metastatic potential of melanoma cells. Despite recent progresses in melanoma targeted therapies, this malignancy still could not be efficiently managed (Zingg et al, 2015). Novel therapeutic strategies have been developed over the past few decades (Hersey and Gallagher, 2012; Niezgoda et al, 2015), metastatic melanoma is associated with a poor prognosis (Tsao et al, 2012). Better therapies for melanoma are in urgent need to be established

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