敲低RAD51AP1可通过抑制CD133阳性卵巢癌干细胞样细胞的自我更新来增强化学敏感性。

Knocking down RAD51AP1 enhances chemosensitivity by inhibiting the self-renewal of CD133 positive ovarian cancer stem-like cells.

作者信息

Zeng Si-Heng, Yan Zhi-Qiang, Ren Qing, Lin Li-Hui, Chen Zhen

机构信息

Department of Gynecology, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University, Shanghai, 200170, China.

Department of Gynecology, Hainan West Central Hospital, Danzhou, 571700, China.

出版信息

Discov Oncol. 2024 Sep 5;15(1):410. doi: 10.1007/s12672-024-01258-9.

DOI:10.1007/s12672-024-01258-9

PMID:39235706

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11377390/

Abstract

PURPOSE

This study was designed to investigate the function of RAD51AP1 in the self-renewal and chemosensitivity of CD133 positive (CD133) ovarian cancer (OC) stem-like cells.

METHODS

CD133 (CD133 positive) OVCAR4 and CD133 negative (CD133) OVCAR4 cells were separated from OVCAR4 by flow cytometry. Then, the separated CD133OVCAR4 cells were divided into the following groups: Vector group; RAD51AP1 group; siNC group; si-RAD51AP1 group. Next, sphere-formation assay and colony forming assay were used to evaluate the self-renewal and proliferation ability of cells; western blot to detect the expression of RAD51AP1, transforming growth factor beta 1 (TGF-β1) and SMAD4 proteins in tissues and cells; qRT-PCR to assess the mRNA levels of sex-determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (OCT4), NANOG and Kruppel-like factor 4 (KLF4).

RESULTS

The performance of CD133OVCAR4 cells was much better than that of CD133OVCAR4 cells in sphere-formation assay and colony forming assay. Besides, compared with adjacent group and CD133OVCAR4 cells, the expression level of RAD51AP1 increased significantly in OC group and CD133OVCAR4 cells. Moreover, the over-expression of RAD51AP1 promoted the self-renewal and proliferation of CD133OVCAR4 cells. On the contrary, knocking down the expression level of RAD51AP1 could inhibit the self-renewal and proliferation of CD133OVCAR4 cells and improve the sensitivity of cells to chemotherapy drugs.

CONCLUSION

The findings of this study showed that RAD51AP1 was highly expressed in OC tissue and CD133OVCAR4 cells, and regulated the self-renewal and chemosensitivity of tumor cells through the TGF-β1/SMAD4 signaling pathway.

摘要

目的

本研究旨在探讨RAD51AP1在CD133阳性（CD133⁺）卵巢癌（OC）干细胞自我更新及化学敏感性中的作用。

方法

采用流式细胞术从OVCAR4细胞中分离出CD133⁺OVCAR4和CD133⁻OVCAR4细胞。然后，将分离出的CD133⁺OVCAR4细胞分为以下几组：载体组；RAD51AP1组；siNC组；si-RAD51AP1组。接下来，采用成球实验和集落形成实验评估细胞的自我更新和增殖能力；蛋白质免疫印迹法检测组织和细胞中RAD51AP1、转化生长因子β1（TGF-β1）和SMAD4蛋白的表达；实时荧光定量聚合酶链反应（qRT-PCR）评估性别决定区Y盒2（SOX2）、八聚体结合转录因子4（OCT4）、NANOG和 Kruppel样因子4（KLF4）的mRNA水平。

结果

在成球实验和集落形成实验中，CD133⁺OVCAR4细胞的表现明显优于CD133⁻OVCAR4细胞。此外，与相邻组和CD133⁻OVCAR4细胞相比，RAD51AP1在OC组和CD133⁺OVCAR4细胞中的表达水平显著升高。而且，RAD51AP1的过表达促进了CD133⁺OVCAR4细胞的自我更新和增殖。相反，敲低RAD51AP1的表达水平可抑制CD133⁺OVCAR4细胞的自我更新和增殖，并提高细胞对化疗药物的敏感性。