CLIC1對膠質母細胞瘤細胞系凋亡影響的實驗研究

【摘要】 目的 研究CLIC1對膠質母細胞瘤（GBM）細胞系U251、T98凋亡的影響。方法 根據癌癥基因組圖譜（TCGA）數據庫數據分析CLIC1相關臨床表達差異、生存曲線等。設計兩條慢病毒序列，穩定敲低U251、T98細胞系中CLIC1表達。平板克隆形成實驗、CCK8細胞增殖實驗評估對照組、敲低組GBM細胞增殖能力。使用流式檢測對照組、敲低組細胞凋亡率。使用免疫熒光、Western Blot檢測對照組、敲低組凋亡相關蛋白變化。裸鼠異種移植模型建立，觀察CLIC1對裸鼠顱內腫瘤生長的影響、腫瘤大小、凋亡及增殖蛋白染色強度，Western Blot檢測鼠腦凋亡蛋白表達量。結果 泛癌分析表明，CLIC1在廣泛腫瘤中較正常組織高表達，包括膠質瘤，且在膠質瘤中CLIC1表達隨級別呈遞增趨勢，高表達CLIC1的患者整體生存期縮短。敲低CLIC1可抑制GBM細胞克隆形成，降低細胞活力，且能增加GBM細胞系凋亡率，增加凋亡蛋白BAX、Cleaved-Caspase3表達，減少抗凋亡蛋白BCL2表達。體內實驗結果表明，CLIC1在維持腫瘤進展中發揮重要作用。結論 敲低CLIC1可抑制GBM細胞增殖，促進GBM細胞凋亡和延緩顱內進展。

【關鍵詞】 膠質母細胞瘤；氯離子胞內通道 1；細胞凋亡

【中圖分類號】 R739.41" 【文獻標志碼】 A" 【文章編號】 1672-7770（2025）01-0047-08

Experimental study on the effect of CLIC1 on apoptosis in glioblastoma cell lines

Abstract： Objective To investigate the effect of CLIC1 on apoptosis in glioblastoma（GBM） cell lines U251 and T98. Methods The clinical expression differences and survival curves related to CLIC1 were analyzed using data from The Cancer Genome Atlas（TCGA） database. Two lentiviral sequences were designed to stably knock down CLIC1 expression in U251 and T98 cell lines. A plate clone formation assay and CCK-8 cell proliferation assay were conducted to evaluate the proliferation ability of GBM cells in control and knockdown groups. The apoptosis rate in these groups was assessed via flow cytometry. Immunofluorescence and Western blotting were employed to detect changes in apoptosis-related protein expression. A nude mouse xenograft model was established to study the impact of CLIC1 on intracranial tumor growth. The tumor size， the intensity of apoptotic and proliferative protein expression were observed. Western blotting was used to detect apoptotic protein expression in the mouse brain." Results Pan-cancer analysis revealed that CLIC1 was significantly overexpressed in various tumors， including gliomas， compared to normal tissues. In gliomas， CLIC1 expression increased with tumor grade， and high CLIC1 expression correlated with shorter overall survival in patients. Knockdown of CLIC1 inhibited GBM cell clone formation， reduced cell viability， and increased the apoptosis rate. Additionally， knockdown enhanced the" expression of pro-apoptotic proteins BAX and Cleaved-Caspase3， while decreasing the anti-apoptotic protein BCL2. In vivo experiments demonstrated that CLIC1 played a critical role in sustaining tumor"progression. Conclusions Knockdown of CLIC1 inhibits GBM cell proliferation， promotes GBM cell apoptosis， and delays intracranial progression.

Key words： glioblastoma; CLIC1; cell apoptosis

膠質母細胞瘤（glioblastoma，GBM）是最常見的原發性惡性腫瘤，大約每10萬人中有4人患病，可占全部原發惡性中樞系統腫瘤的50%［1］。GBM具有高病死率、高致殘率、高復發率，GBM患者目前采用的標準治療方案是以手術治療為主，術后輔以放化療［2］。目前最常用的化療方案是熟知的Stupp方案，但是對于O6-甲基鳥嘌呤-DNA甲基轉移酶（O6-methylguanine-DNA methyltransferase，MGMT）未甲基化的GBM患者，整體生存率并沒有明顯的延長［1］。……