hsa-miR-301a-3p 通过调控 RUNX3 抑制自然杀伤细胞对非小细胞肺癌细胞的杀伤作用。

Hsa-miR-301a-3p inhibited the killing effect of natural killer cells on non-small cell lung cancer cells by regulating RUNX3.

出版信息

Cancer Biomark. 2023;37(4):249-259. doi: 10.3233/CBM-220469.

DOI:10.3233/CBM-220469

PMID:37302028

Abstract

BACKGROUND

Non-small cell lung cancer (NSCLC) is the most commonly diagnosed solid tumor. Natural killer (NK) cell-based immunotherapy is a promising anti-tumor strategy in various cancers including NSCLC.

OBJECTIVE

We aimed to investigate the specific mechanisms that regulate the killing effect of NK cells to NSCLC cells.

METHODS

Reverse transcription-quantitative PCR (RT-qPCR) assay was applied to measure the levels of hsa-microRNA (miR)-301a-3p and Runt-related transcription factor 3 (RUNX3). Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of IFN-γ and TNF-α. Lactate dehydrogenase assay was applied to detect the killing effect of NK cells. Dualluciferase reporter assay and RNA immunoprecipitation (RIP) assay were carried out to confirm the regulatory relationship between hsa-miR-301a-3p and RUNX3.

RESULTS

A low expression of hsa-miR-301a-3p was observed in NK cells stimulated by IL-2. The levels of IFN-γ and TNF-α were increased in NK cells of the IL-2 group. Overexpression of hsa-miR-301a-3p reduced the levels of IFN-γ and TNF-α as well as the killing effect of NK cells. Furthermore, RUNX3 was identified to be a target of hsamiR-301a-3p. hsa-miR-301a-3p suppressed the cytotoxicity of NK cells to NSCLC cells by inhibiting the expression of RUNX3. We found hsa-miR-301a-3p promoted tumor growth by suppressing the killing effect of NK cells against NSCLC cells in vivo.

CONCLUSIONS

Hsa-miR-301a-3p suppressed the killing effect of NK cells on NSCLC cells by targeting RUNX3, which may provide promising strategies for NK cell-based antitumor therapies.

摘要

背景

非小细胞肺癌（NSCLC）是最常见的实体肿瘤。自然杀伤（NK）细胞为基础的免疫疗法是一种很有前途的抗肿瘤策略，在包括 NSCLC 在内的各种癌症中。

目的

我们旨在研究调节 NK 细胞对 NSCLC 细胞杀伤作用的具体机制。

方法

采用逆转录定量 PCR（RT-qPCR）法检测 hsa-微小 RNA（miR）-301a-3p 和 runt 相关转录因子 3（RUNX3）的水平。酶联免疫吸附试验（ELISA）用于检测 IFN-γ 和 TNF-α 的水平。乳酸脱氢酶试验用于检测 NK 细胞的杀伤作用。双荧光素酶报告基因实验和 RNA 免疫沉淀（RIP）实验用于证实 hsa-miR-301a-3p 与 RUNX3 之间的调控关系。

结果

IL-2 刺激的 NK 细胞中 hsa-miR-301a-3p 的表达水平较低。IL-2 组 NK 细胞中 IFN-γ 和 TNF-α 的水平增加。hsa-miR-301a-3p 的过表达降低了 NK 细胞中 IFN-γ 和 TNF-α 的水平以及 NK 细胞的杀伤作用。此外，RUNX3 被鉴定为 hsa-miR-301a-3p 的靶标。hsa-miR-301a-3p 通过抑制 RUNX3 的表达来抑制 NK 细胞对 NSCLC 细胞的细胞毒性。我们发现 hsa-miR-301a-3p 通过抑制 NK 细胞对 NSCLC 细胞的杀伤作用促进了肿瘤的生长。