BCLX前體mRNA剪接修飾單獨或聯合放療對人腦膠質母細胞瘤細胞具有抗腫瘤作用

2024年2月，中國科學院近代物理研究所生物醫學研究中心，先進能源科學技術廣東省重點實驗室，中國科學院重離子輻射生物學與醫學重點實驗室，中國科學院大學生命科學學院

(Bio-Medical Research Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, China.

Advanced Energy Science and Technology Guangdong Laboratory, uizhou 516029, China.

Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences, Lanzhou 730000, China.

College of Life Sciences, University of Chinese Academy of Sciences, Beijing 101408, China. ） Cuixia Di研究團隊，在《CDD press》上發表論文，標題為：“Modification of BCLX pre-mRNA splicing has antitumor efficacy alone or in combination with radiotherapy in human glioblastoma cells”

“BCLX前體mRNA剪接修飾單獨或聯合放療對人腦膠質母細胞瘤細胞具有抗腫瘤作用”

Abstract

Dysregulation of anti-apoptotic and pro-apoptotic protein isoforms arising from aberrant splicing is a crucial hallmark of cancers and may contribute to therapeutic resistance. Thus, targeting RNA splicing to redirect isoform expression of apoptosis-related genes could lead to promising anti-cancer phenotypes. Glioblastoma (GBM) is the most common type of malignant brain tumor in adults. In this study, through RT-PCR and Western Blot analysis, we found that BCLX pre-mRNA is aberrantly spliced in GBM cells with a favored splicing of anti-apoptotic Bcl-xL. Modulation of BCLX pre-mRNA splicing using splice-switching oligonucleotides (SSOs) efficiently elevated the pro-apoptotic isoform Bcl-xS at the expense of the anti-apoptotic Bcl-xL. Induction of Bcl-xS by SSOs activated apoptosis and autophagy in GBM cells. In addition, we found that ionizing radiation could also modulate the alternative splicing of BCLX. In contrast to heavy (carbon) ion irradiation, low energy X-ray radiation-induced an increased ratio of Bcl-xL/Bcl-xS. Inhibiting Bcl-xL through splicing regulation can significantly enhance the radiation sensitivity of 2D and 3D GBM cells. These results suggested that manipulation of BCLX pre-mRNA alternative splicing by splice-switching oligonucleotides is a novel approach to inhibit glioblastoma tumorigenesis alone or in combination with radiotherapy.

摘要

由異常剪接引起的抗凋亡和促凋亡蛋白異構體的失調是癌癥的一個重要標志，并可能導致治療耐藥性。因此，靶向RNA剪接來重定向凋亡相關基因的異構體表達可能會導致有希望的抗癌表型。膠質母細胞瘤(GBM)是成人最常見的惡性腦腫瘤。在本研究中，我們通過RT-PCR和Western Blot分析發現，BCLX pre-mRNA在GBM細胞中存在異常剪接，其中抗凋亡的Bcl-xL剪接更有利。使用剪接開關寡核苷酸(SSOs)調節BCLX pre-mRNA剪接可以有效地提高促凋亡異構體Bcl-xS，而降低抗凋亡的Bcl-xL。SSOs誘導Bcl-xS可激活GBM細胞凋亡和自噬。此外，我們發現電離輻射也可以調節BCLX的選擇性剪接。與重(碳)離子輻照相比，低能x射線輻照誘導Bcl-xL/Bcl-xS的比值增加。通過剪接調控抑制Bcl-xL可顯著增強二維和三維GBM細胞的輻射敏感性。這些結果表明，通過剪接開關寡核苷酸操縱BCLX前mrna選擇性剪接是一種單獨或聯合放療抑制膠質母細胞瘤腫瘤發生的新方法。

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