Qiang Jing, Zhao Cheng, Shi Liu-Qing, Sun Si-Rui, Wang Hua-Kai, Liu Shi-Lei, Yang Zi-Yi, Dong Ping, Xiang Shan-Shan, Wang Jian-Dong, Shu Yi-Jun
Laboratory of General Surgery and Department of General Surgery, Xinhua Hospital affiliated with Shanghai Jiao Tong University School of Medicine, No. 1665 Kongjiang Road, Shanghai, 200092, China.
Shanghai Key Laboratory of Biliary Tract Disease Research, Shanghai, China, No. 1665 Kongjiang Road, Shanghai, 200092, China.
Gene Ther. 2024 Nov;31(11-12):594-606. doi: 10.1038/s41434-024-00488-4. Epub 2024 Sep 21.
Gallbladder cancer (GBC) is highly aggressive and has poor prognosis, with most patients only diagnosed at an advanced stage. Furthermore, treatment options are limited, and their effect is unsatisfactory. Bromodomain-containing protein (BRD) is an epigenetic regulator that plays a carcinogenic role in several tumors, including squamous cell lung cancer, acute myeloid leukemia, synovial sarcoma, and malignant rhabdomyosarcoma. However, the expression, biological function, and molecular mechanisms of action of BRD9 in GBC are still unknown. Kaplan-Meier analysis, qRT-PCR, and analysis of clinical features were used to assess the clinical significance of BRD9 in GBC. Cell Counting Kit-8 and colony formation assays were performed to determine the effects of BRD9 on cell growth. The functional role of BRD9 in GBC was explored using qRT-PCR, western blotting, siRNA, and CHIP-qPCR. mRNA sequencing was performed to explore the underlying mechanisms of BRD9, and a nude mouse model of GBC was established to explore the anti-tumor effects of the BRD9 inhibitor I-BRD9 in vivo. BRD9 expression was elevated in GBC tissues compared with adjacent non-tumor tissues, and high BRD9 expression was associated with poor prognosis in patients with GBC. BRD9 knockdown by siRNA significantly decreased cell growth. Targeting BRD9 with I-BRD9 inhibited the proliferation of GBC cells without significant toxic effects. Additionally, I-BRD9 treatment suppressed CST1 expression in GBC cell lines, thereby inhibiting the PI3K-AKT pathway. The transcription factor FOXP1 was found to interact with BRD9 to regulate CST1 expression. Collectively, these results suggest that BRD9 may be a promising biomarker and therapeutic target for GBC.
胆囊癌(GBC)具有高度侵袭性且预后较差,大多数患者在晚期才被诊断出来。此外,治疗选择有限,且效果不尽人意。含溴结构域蛋白(BRD)是一种表观遗传调节因子,在包括肺鳞状细胞癌、急性髓系白血病、滑膜肉瘤和恶性横纹肌肉瘤在内的多种肿瘤中发挥致癌作用。然而,BRD9在GBC中的表达、生物学功能及分子作用机制仍不清楚。采用Kaplan-Meier分析、qRT-PCR及临床特征分析来评估BRD9在GBC中的临床意义。进行细胞计数试剂盒-8及集落形成试验以确定BRD9对细胞生长的影响。利用qRT-PCR、蛋白质免疫印迹法、小干扰RNA(siRNA)及染色质免疫沉淀-定量聚合酶链反应(CHIP-qPCR)探究BRD9在GBC中的功能作用。进行信使核糖核酸(mRNA)测序以探究BRD9的潜在机制,并建立GBC裸鼠模型以探究BRD9抑制剂I-BRD9在体内的抗肿瘤作用。与癌旁非肿瘤组织相比,GBC组织中BRD9表达升高,且BRD9高表达与GBC患者的不良预后相关。通过siRNA敲低BRD9可显著降低细胞生长。用I-BRD9靶向BRD9可抑制GBC细胞增殖且无明显毒性作用。此外,I-BRD9处理可抑制GBC细胞系中 CST1的表达,从而抑制磷脂酰肌醇-3激酶-蛋白激酶B(PI3K-AKT)通路。发现转录因子叉头框蛋白P1(FOXP1)与BRD9相互作用以调节CST1表达。总体而言,这些结果表明BRD9可能是GBC一个有前景的生物标志物和治疗靶点。