Wang Tianqing, Xin Chuan, Zhang Shiyu, Tian Xin, Hu Yuting, Wang Ying, Wang Jiongke, Ji Ning, Zeng Xin, Li Jing
State Key Laboratory of Oral Diseases & National Center for Stomatology, National Clinical Research Center for Oral Diseases, Research Unit of Oral Carcinogenesis and Management, Chinese Academy of Medical Sciences, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China.
Key Laboratory of Oral Biomedical Research of Zhejiang Province, School of Stomatology, Zhejiang University School of Medicine, Zhejiang Provincial Clinical Research Center for Oral Diseases, Stomatology Hospital, Cancer Center of Zhejiang University, Hangzhou 310006, China.
Cancers (Basel). 2023 Nov 9;15(22):5351. doi: 10.3390/cancers15225351.
Chemoresistance poses a significant challenge in the treatment of advanced head and neck squamous cell cancer (HNSCC). The role and mechanism of circular RNAs (circRNAs) in HNSCC chemoresistance remain understudied. We conducted circRNA microarray analysis to identify differentially expressed circRNAs in HNSCC. The expression of circRNAs from the tyrosylprotein sulfotransferase 2 (TPST2) gene and miRNAs was evaluated through qPCR, while the circular structure of circTPST2 was verified using Sanger sequencing and RNase R. Through Western blotting, biotin-labeled RNA pulldown, RNA immunoprecipitation, mass spectrometry, and rescue experiments, we discovered miR-770-5p and nucleolin as downstream targets of circTPST2. Functional tests, including CCK8 assays and flow cytometry, assessed the chemoresistance ability of circTPST2, miR-770-5p, and Nucleolin. Additionally, FISH assays determined the subcellular localization of circTPST2, miR-770-5p, and Nucleolin. IHC staining was employed to detect circTPST2 and Nucleolin expression in HNSCC patients. circTPST2 expression was inversely correlated with cisplatin sensitivity in HNSCC cell lines. Remarkably, high circTPST2 expression correlated with lower overall survival rates in chemotherapeutic HNSCC patients. Mechanistically, circTPST2 reduced chemosensitivity through sponge-like adsorption of miR-770-5p and upregulation of the downstream protein Nucleolin in HNSCC cells. The TCGA database revealed improved prognosis for patients with low circTPST2 expression after chemotherapy. Moreover, analysis of HNSCC cohorts demonstrated better prognosis for patients with low Nucleolin protein expression after chemotherapy. We unveil circTPST2 as a circRNA associated with chemoresistance in HNSCC, suggesting its potential as a marker for selecting chemotherapy regimens in HNSCC patients. Further exploration of the downstream targets of circTPST2 advanced our understanding and improved treatment strategies for HNSCC.
化疗耐药性是晚期头颈部鳞状细胞癌（HNSCC）治疗中的一个重大挑战。环状RNA（circRNA）在HNSCC化疗耐药中的作用和机制仍未得到充分研究。我们进行了circRNA微阵列分析，以鉴定HNSCC中差异表达的circRNA。通过qPCR评估了来自酪氨酰蛋白磺基转移酶2（TPST2）基因的circRNA和miRNA的表达，同时使用桑格测序和RNase R验证了circTPST2的环状结构。通过蛋白质免疫印迹、生物素标记的RNA下拉、RNA免疫沉淀、质谱分析和挽救实验，我们发现miR-770-5p和核仁素是circTPST2的下游靶点。包括CCK8分析和流式细胞术在内的功能测试评估了circTPST2、miR-770-5p和核仁素的化疗耐药能力。此外，荧光原位杂交分析确定了circTPST2、miR-770-5p和核仁素的亚细胞定位。免疫组化染色用于检测HNSCC患者中circTPST2和核仁素的表达。circTPST2的表达与HNSCC细胞系中的顺铂敏感性呈负相关。值得注意的是，在接受化疗的HNSCC患者中，circTPST2高表达与较低的总生存率相关。机制上，circTPST2通过海绵样吸附miR-770-5p并上调HNSCC细胞中的下游蛋白核仁素来降低化疗敏感性。TCGA数据库显示，化疗后circTPST2低表达的患者预后较好。此外，对HNSCC队列的分析表明，化疗后核仁素蛋白低表达的患者预后较好。我们揭示了circTPST2是一种与HNSCC化疗耐药相关的circRNA，表明其作为HNSCC患者化疗方案选择标志物的潜力。对circTPST2下游靶点的进一步探索增进了我们对HNSCC的理解，并改善了治疗策略。
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