南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (10): 1440-1451.doi: 10.12122/j.issn.1673-4254.2022.10.02

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CircPCSK5在胃癌中高表达并促进胃癌细胞的增殖、侵袭和上皮间质转化

左学良，蔡 娟，陈志强，李艳娜，张斗峰   

  1. 皖南医学院第一附属医院（弋矶山医院）胃肠外科，重大疾病非编码RNA转化研究安徽普通高校重点实验室，肿瘤内科，安徽 芜湖 241001；南京医科大学第一附属医院肝胆中心，江苏 南京 210029
  • 出版日期:2022-10-20 发布日期:2022-11-01

CircPCSK5 is highly expressed in gastric cancer and promotes cancer cell proliferation, invasion and epithelial-mesenchymal transition

ZUO Xueliang, CAI Juan, CHEN Zhiqiang, LI Yanna, ZHANG Doufeng   

  1. Department of Gastrointestinal Surgery, Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institution, Department of Oncology, First Affiliated Hospital (Yijishan Hospital) of Wannan Medical College, Wuhu 241001, China; Hepatobiliary Center, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Online:2022-10-20 Published:2022-11-01

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摘要： 目的 探讨circPCSK5在胃癌中的表达情况和对胃癌细胞增殖、侵袭和上皮间质转化的影响。方法 通过高通量测序构建胃癌组织和癌旁正常胃粘膜组织的circRNA差异表达谱。选取在皖南医学院第一附属医院行胃癌根治术的患者共62例作为研究对象，利用RT-qPCR检测circPCSK5在胃癌组织和细胞系中的表达情况。采用χ2检验分析circPCSK5表达水平与胃癌临床病理资料相关性，利用Kaplan?Meier法绘制患者的总生存和无病生存曲线，Cox比例风险回归模型分析影响胃癌患者预后的独立危险因素。通过RNase R和actinomycin D实验检测circPCSK5的稳定性。利用荧光原位杂交和细胞核质分离实验检测circPCSK5的亚细胞定位。通过CCK-8和EdU实验检测circPCSK5对胃癌细胞增殖能力的影响，通过transwell实验检测circPCSK5对胃癌细胞迁移和侵袭能力的影响。利用Western blot和RT-qPCR检测敲低或过表达circPCSK5后胃癌细胞上皮-间质转化标记物的表达变化。结果 在胃癌组织和细胞中，circPCSK5的表达明显升高（P<0.001，P<0.01）。CircPCSK5表达水平与患者的肿瘤大小、脉管侵犯、淋巴结转移和AJCC分期存在明显正相关性（P<0.05）。CircPCSK5高表达患者的总生存和无病生存时间明显低于circPCSK5低表达患者（P<0.001），且circPCSK5高表达是影响胃癌患者预后的独立危险因素（P<0.05）。敲低circPCSK5表达能明显抑制HGC27细胞的增殖、迁移和侵袭能力（P<0.01），并导致上皮间质转化标记物E-cadherin表达升高，N-cadherin和Vimentin表达降低（P<0.01）。过表达circPCSK5能促进MKN45细胞的增殖、迁移和侵袭能力（P<0.01），降低E-cadherin表达，升高N-cadherin和Vimentin表达（P<0.01）。结论 CircPCSK5在胃癌中高表达，并能促进胃癌细胞的增殖、侵袭和上皮间质转化，可作为胃癌预后预测指标和潜在的分子治疗靶点。

关键词: 环状RNA；胃癌；预后；增殖；侵袭；上皮间质转化

Abstract: Objective To investigate the expression of circPCSK5 in gastric cancer (GC) and its role in regulation of theproliferation, invasion and epithelial-mesenchymal transition (EMT) of GC cells. Methods High-throughput sequencing was performed in 3 pairs of GC and adjacent gastric mucosa tissues to obtain the differential expression profile of circRNA. The expression of circPCSK5 was detected in 62 patients undergoing radical surgery for GC using RT-qPCR, and the correlation between circPCSK5 expression level and clinicopathological data of the patients was analyzed. The overall survival and disease- free survival of the patients were assessed with Kaplan-Meier survival analysis, and the independent risk factors affecting the patients' prognosis were analyzed using Cox proportional hazards regression model. The stability and subcellular localization of circPCSK5 were assessed using RNase R and actinomycin D assays, fluorescence in situ hybridization and nucleocytoplasmic separation assay. CCK-8 assay, EdU assay and Transwell assay were employed to examine the changes in proliferation, migration and invasion of GC cells with circPCSK5 knockdown or overexpression; Western blotting and RT-qPCR assays were used to detect the expression levels of EMT markers in the transfected cells. Results The expression of circPCSK5 was significantly upregulated in GC tissues and cells (P<0.001, P<0.01). The expression level of circPCSK5 was positively correlated with tumor size, vascular invasion, lymph node metastasis and AJCC stage of GC (P<0.05). The overall survival and disease-free survival were significantly lower in GC patients with high circPCSK5 expression than in those with low circPCSK5 expression (P<0.001). High circPCSK5 expression was an independent risk factor for a poor prognosis of GC patients (P<0.05). Knockdown of circPCSK5 significantly inhibited the proliferation, migration and invasion of HGC27 cells (P<0.01), increased the expressions of E-cadherin, and decreased the expression of N-cadherin and vimentin (P<0.01). CircPCSK5 overexpression promoted the proliferation, migration and invasion of MKN45 cells (P<0.01), reduced E-cadherin expression and increased N-cadherin and vimentin expressions (P<0.01). Conclusion CircPCSK5 is highly expressed in GC and promotes the proliferation, invasion and EMT of GC cells, suggesting its potential as a prognostic biomarker and therapeutic target for GC.

Key words: circular RNA; gastric cancer; prognosis; proliferation; invasion; epithelial-mesenchymal transition