miR-342-3p 通过 HDAC7/PTEN 轴抑制 LCSC 致癌性和细胞干性。

MiR-342-3p inhibits LCSC oncogenicity and cell stemness through HDAC7/PTEN axis.

机构信息

Department of Infectious Disease (No. 3), Second Affiliated Hospital of Harbin Medical University, Harbin, 150040, Heilongjiang, People's Republic of China.

Department of Respiratory Oncology, Harbin Medical University Cancer Hospital, No. 150, Haping Road, Nangang District, Harbin, 150040, Heilongjiang, People's Republic of China.

出版信息

Inflamm Res. 2022 Jan;71(1):107-117. doi: 10.1007/s00011-021-01521-7. Epub 2021 Nov 29.

DOI:10.1007/s00011-021-01521-7

PMID:34842937

Abstract

OBJECTIVE

This study aims to explore the effects of miR-342-3p on liver cancer stem cells (LCSC) and related mechanism.

METHODS

LCSC were sorted using immunomagnetic beads and flow cytometry was used to determine CD133+ and CD133- sorted cells. The self-renewal ability and growth ability of LCSC were measured by tumor spheroid formation assay and soft agar colony formation assay. Protein and mRNA expressions of CD44, ALDH1, Bmi1, Sox2 and Oct4 were detected by western blot and quantitative PCR. The relationship between miR-342-3p and HDAC7 was analyzed by dual-luciferase assay. The acetylation level of H3 protein was measured by acetyl Lysine antibody.

RESULTS

miR-342-3p overexpression in LCSC lead to lower tumor volume, reduced tumor spheroid formation and agar colony formation rates, as well as lower mRNA and protein expressions of CD44, ALDH1, Bmi1, Sox2, and Oct4. Dual-luciferase reporter assay confirmed HDAC7 as a target gene of miR-342-3p. Inhibition of HDAC7 or overexpression of PTEN suppressed the carcinogenicity and stemness of LCSC. PTEN expression was increased in sh-HDAC7 group and decreased in pcDNA3.1-HDAC7 group. HDAC7 promoted H3 deacetylation and inhibited PTEN expression. Overexpression of HDAC7 or silencing of PTEN could reverse the inhibitory effect of overexpression of miR-342-3p on LCSC carcinogenicity and cell stemness.

CONCLUSION

MiR-342-3p inhibited LCSC oncogenicity and cell stemness by promoting PTEN and inhibiting HDAC7.

摘要

目的

本研究旨在探讨 miR-342-3p 对肝癌干细胞（LCSC）的影响及其相关机制。

方法

采用免疫磁珠法分选 LCSC，流式细胞术检测 CD133+和 CD133-分选细胞。采用肿瘤球形成实验和软琼脂集落形成实验检测 LCSC 的自我更新能力和生长能力。采用 Western blot 和定量 PCR 检测 CD44、ALDH1、Bmi1、Sox2 和 Oct4 的蛋白和 mRNA 表达。采用双荧光素酶报告实验分析 miR-342-3p 与 HDAC7 的关系。采用乙酰化赖氨酸抗体检测 H3 蛋白的乙酰化水平。

结果

miR-342-3p 在 LCSC 中的过表达导致肿瘤体积减小，肿瘤球形成和琼脂集落形成率降低，以及 CD44、ALDH1、Bmi1、Sox2 和 Oct4 的 mRNA 和蛋白表达降低。双荧光素酶报告实验证实 HDAC7 是 miR-342-3p 的靶基因。抑制 HDAC7 或过表达 PTEN 可抑制 LCSC 的致癌性和干细胞特性。在 sh-HDAC7 组中，PTEN 表达增加，在 pcDNA3.1-HDAC7 组中，PTEN 表达降低。HDAC7 促进 H3 去乙酰化并抑制 PTEN 表达。HDAC7 过表达或 PTEN 沉默可逆转 miR-342-3p 过表达对 LCSC 致癌性和细胞干性的抑制作用。