| 摘要: |
| 目的 探究环状RNA circNEIL3在乳腺癌中的表达、生物学功能及其对细胞周期相关蛋白表达的影响。方法 收集50例经病理证实的乳腺癌患者的临床病理资料及乳腺癌组织和癌旁正常组织样本。采用环状RNA测序分析3对乳腺癌组织与癌旁正常组织的差异表达环状RNA，利用RNA酶消化实验和核质分离实验验证差异表达环状RNA circNEIL3的特征。采用qPCR检测circNEIL3在乳腺癌组织和细胞中的表达，分析circNEIL3表达与患者临床病理资料的关系。采用CCK-8实验和集落形成实验检测circNEIL3对BT-549和MCF-7细胞增殖的影响，运用流式细胞术检测细胞凋亡及细胞周期变化，采用蛋白质印迹法检测细胞周期蛋白D1（CCND1）、细胞周期蛋白依赖性激酶（CDK）4、细胞周期蛋白E1（CCNE1）和CDK2蛋白的表达水平。结果 乳腺癌组织与癌旁正常组织中差异表达的circNEIL3不易被RNA酶R降解，其主要分布于乳腺癌细胞的细胞质中。与癌旁正常乳腺组织及细胞相比，乳腺癌组织和细胞中circNEIL3的表达水平均增高（均P＜0.05）。肿瘤最大径＞2 cm和高TNM 分期（Ⅲ期）的乳腺癌患者多表现为circNEIL3 高表达（均P＜0.05）。circNEIL3过表达可增强乳腺癌细胞增殖活性，并上调CCND1、CDK4、CCNE1和CDK2的蛋白表达水平（均P＜0.05）。敲低circNEIL3可诱导乳腺癌细胞凋亡（P＜0.01），导致细胞周期阻滞于G1期（P＜0.01）。结论 乳腺癌组织和细胞中高表达的circNEIL3可通过调控细胞周期相关蛋白的表达促进乳腺癌细胞增殖。 |
| 关键词: 乳腺肿瘤 circNEIL3 细胞增殖 细胞凋亡 细胞周期 |
| DOI：10.16781/j.CN31-2187/R.20220230 |
| 投稿时间：2022-03-18修订日期：2022-07-01 |
| 基金项目: |
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| High expression of circular RNA circNEIL3 promotes proliferation of breast cancer cells |
| CHEN Hang1,2,LI Molin3,CHEN Yi2* |
(1. Department of Cell Biology and Genetics, School of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China;
2. Department of Surgery (Ⅱ), The Central Hospital of Wanyuan, Dazhou 636350, Sichuan, China;
3. Laboratory of Bone Development and Regeneration, Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, China
*Corresponding author) |
| Abstract: |
| Objective To explore the expression and function of circular RNA (circRNA) circNEIL3 in breast cancer (BC) and its influence on the expression levels of cell cycle-related proteins. Methods Clinical and pathological data from 50 patients with pathologically confirmed BC, along with samples of their BC tissues and para-cancer tissues were collected. circRNA sequencing was used to analyze the differentially expressed circRNAs in 3 pairs of BC tissues and para-cancer tissues. The characteristic of differentially expressed circRNA circNEIL3 was verified by ribonuclease (RNase) digestion and cytoplasmic/nuclear fractionation experiments. The expression of circNEIL3 in BC tissues and cells was detected by quantitative polymerase chain reaction. The relationship between circNEIL3 expression and the patients’ clinicopathological data was also analyzed. The effect of circNEIL3 on proliferation of BT-549 and MCF-7 cells was identified by cell counting kit 8 and colony-formation assays. The changes of cell apoptosis and cell cycle were detected by flow cytometry. The expression levels of cyclin D1 (CCND1), cyclin-dependent kinase (CDK) 4, cyclin E1 (CCNE1) and CDK2 were measured by Western blotting. Results circNEIL3 differentially expressed in BC tissues and adjacent tissues was not easily degraded by RNase R and mainly located in the cytoplasm of BC cells. Compared with normal breast tissues and cells, the expression levels of circNEIL3 in BC tissues and cells were significantly up-regulated (all P＜0.05). circNEIL3 was highly expressed in BC patients with tumor maximum diameter＞2 cm and at high TNM stage (stage Ⅲ) (both P＜0.05). Overexpression of circNEIL3 enhanced proliferation of BC cells and up-regulated the levels of CCND1, CDK4, CCNE1 and CDK2 (all P＜0.05). Knockdown of circNEIL3 induced cell apoptosis (P＜0.01), resulting in cell cycle arrest at G1 phase (P＜0.01). Conclusion High expression of circNEIL3 in BC tissues and cells can promote the proliferation of BC cells by up-regulating the expression levels of cycle-related proteins. |
| Key words: breast neoplasms circNEIL3 cell proliferation apoptosis cell cycle |
