BCLX前體mRNA剪接修飾單獨(dú)或聯(lián)合放療對(duì)人腦膠質(zhì)母細(xì)胞瘤細(xì)胞具有抗腫瘤作用

2024年2月，中國(guó)科學(xué)院近代物理研究所生物醫(yī)學(xué)研究中心，先進(jìn)能源科學(xué)技術(shù)廣東省重點(diǎn)實(shí)驗(yàn)室，中國(guó)科學(xué)院重離子輻射生物學(xué)與醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室，中國(guó)科學(xué)院大學(xué)生命科學(xué)學(xué)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(tuán)隊(duì)，在《CDD press》上發(fā)表論文，標(biāo)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(dú)或聯(lián)合放療對(duì)人腦膠質(zhì)母細(xì)胞瘤細(xì)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(gòu)體的失調(diào)是癌癥的一個(gè)重要標(biāo)志，并可能導(dǎo)致治療耐藥性。因此，靶向RNA剪接來(lái)重定向凋亡相關(guān)基因的異構(gòu)體表達(dá)可能會(huì)導(dǎo)致有希望的抗癌表型。膠質(zhì)母細(xì)胞瘤(GBM)是成人最常見(jiàn)的惡性腦腫瘤。在本研究中，我們通過(guò)RT-PCR和Western Blot分析發(fā)現(xiàn)，BCLX pre-mRNA在GBM細(xì)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(kāi)關(guān)寡核苷酸(SSOs)調(diào)節(jié)BCLX pre-mRNA剪接可以有效地提高促凋亡異構(gòu)體Bcl-xS，而降低抗凋亡的Bcl-xL。SSOs誘導(dǎo)Bcl-xS可激活GBM細(xì)胞凋亡和自噬。此外，我們發(fā)現(xiàn)電離輻射也可以調(diào)節(jié)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(dǎo)Bcl-xL/Bcl-xS的比值增加。通過(guò)剪接調(diào)控抑制Bcl-xL可顯著增強(qiáng)二維和三維GBM細(xì)胞的輻射敏感性。這些結(jié)果表明，通過(guò)剪接開(kāi)關(guān)寡核苷酸操縱BCLX前mrna選擇性剪接是一種單獨(dú)或聯(lián)合放療抑制膠質(zhì)母細(xì)胞瘤腫瘤發(fā)生的新方法。

該論文中，從人膠質(zhì)母細(xì)胞瘤細(xì)胞系A(chǔ)172、T98G、U251、U87、GSCs(人膠質(zhì)瘤干細(xì)胞樣細(xì)胞)和正常人星形膠質(zhì)細(xì)胞細(xì)胞系HA1800體外培養(yǎng)是使用Ausbian特級(jí)胎牛血清完成的。欲了解或購(gòu)買Ausbian特級(jí)胎牛血清可以聯(lián)系北京締一生物400-166-8600.