LncRNA ZNF649-AS1 promotes trastuzumab resistance and TAM-dependent PD-L1 expression in breast cancer by regulating EXOC7 alternative splicing

IF 3.8 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Archives of biochemistry and biophysics Pub Date : 2024-08-17 DOI:10.1016/j.abb.2024.110128

Huaying Dong , Jing Han , Xiang Chen , Hening Sun , Mingli Han , Wei Wang

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Abstract

Background

Trastuzumab resistance is a serious clinical problem in the treatment of HER2-positive breast cancer (BC). The lncRNA ZNF649-AS1 was previously found to promote HER2-positive BC trastuzumab resistance. The study aims to explore the molecular mechanism of ZNF649-AS1 in HER2-positive BC trastuzumab resistance.

Methods

Tumor tissue and peripheral blood samples were collected from 20 HER2-positive BC patients with trastuzumab-resistant and non-resistant, respectively. Trastuzumab-resistant BC cell lines SKBR-3-TR and BT474-TR were established. RIP was employed to confirm the binding of ZNF649-AS1, PRPF8 and exocyst complex component 7 (EXOC7). RNA expression of EXOC7-L (Full length of EXOC7) and EXOC7-S (Spliceosome of EXOC7) were detected using agarose gel electrophoresis. Expressions of macrophage markers CD68⁺ CD206⁺ were measured by flow cytometry.

Results

ZNF649-AS1 expression was upregulated in HER2-positive BC trastuzumab resistance. ZNF649-AS1 downregulation inhibited trastuzumab resistance in HER2-positive BC. ZNF649-AS1 regulated EXOC7 alternative splicing by binding with PRPF8. EXOC7-S knockdown suppressed trastuzumab resistance and TAM-dependent PD-L1 expression in HER2-positive BC. EXOC7-S overexpression abolished the effects of ZNF649-AS1 knockdown on trastuzumab resistance and TAM-dependent PD-L1 expression in HER2-positive BC.

Conclusion

ZNF649-AS1 promoted trastuzumab resistance and TAM-dependent PD-L1 expression in HER2-positive BC via promoting alternative splicing of EXOC7 by PRPF8.

Abstract Image

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LncRNA ZNF649-AS1通过调节EXOC7的替代剪接促进乳腺癌中曲妥珠单抗的耐药性和TAM依赖性PD-L1的表达。

背景曲妥珠单抗耐药是治疗HER2阳性乳腺癌（BC）的一个严重临床问题。以前曾发现lncRNA ZNF649-AS1可促进HER2阳性BC曲妥珠单抗耐药。本研究旨在探讨ZNF649-AS1在HER2阳性BC曲妥珠单抗耐药中的分子机制：方法：收集20例HER2阳性BC曲妥珠单抗耐药和非耐药患者的肿瘤组织和外周血样本。建立了曲妥珠单抗耐药的BC细胞系SKBR-3-TR和BT474-TR。采用 RIP 法确认 ZNF649-AS1、PRPF8 和外囊复合体成分 7（EXOC7）的结合。琼脂糖凝胶电泳检测了 EXOC7-L（EXOC7 全长）和 EXOC7-S（EXOC7 的剪接体）的 RNA 表达。流式细胞术检测了巨噬细胞标志物 CD68+ CD206+ 的表达：结果：ZNF649-AS1在HER2阳性的曲妥珠单抗耐药的BC中表达上调。下调ZNF649-AS1可抑制HER2阳性BC曲妥珠单抗耐药。ZNF649-AS1通过与PRPF8结合调控EXOC7的替代剪接。EXOC7-S敲除抑制了HER2阳性BC的曲妥珠单抗抗性和TAM依赖性PD-L1表达。在HER2阳性BC中，EXOC7-S过表达可消除ZNF649-AS1敲除对曲妥珠单抗抗性和TAM依赖性PD-L1表达的影响：结论：ZNF649-AS1通过PRPF8促进EXOC7的替代剪接，促进HER2阳性BC的曲妥珠单抗耐药和TAM依赖性PD-L1表达。

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来源期刊

Archives of biochemistry and biophysics 生物-生化与分子生物学

CiteScore

7.40

自引率

0.00%

发文量

245

审稿时长

26 days

期刊介绍： Archives of Biochemistry and Biophysics publishes quality original articles and reviews in the developing areas of biochemistry and biophysics. Research Areas Include: • Enzyme and protein structure, function, regulation. Folding, turnover, and post-translational processing • Biological oxidations, free radical reactions, redox signaling, oxygenases, P450 reactions • Signal transduction, receptors, membrane transport, intracellular signals. Cellular and integrated metabolism.