RRM2在非小细胞肺癌中生物学行为的研究

Biological Behavior of RRM2 in Non-Small Cell Lung Cancer

摘要

目的 探讨核糖核苷酸还原酶调节亚基M2（RRM2）在非小细胞肺癌（NSCLC）中的表达和生物学行为。方法 从癌症基因组图谱（TCGA）数据库中获取NSCLC患者的基因表达数据和临床数据,分析RRM2基因在正常组织和NSCLC组织中的表达,利用NSCLC患者的临床数据绘制生存曲线;实时荧光定量PCR（qPCR）检测RRM2基因在细胞系中的表达情况以及验证RRM2基因在细胞系中的沉默和过表达效率;蛋白质免疫印迹法检测RRM2基因在细胞系中的蛋白表达,以及验证其在细胞系中沉默和过表达效率;Cell Counting Kit-8（CCK-8）、流式细胞术和Transwell实验检测细胞的增值、凋亡、侵袭和迁移能力。结果 RRM2在肺鳞状细胞癌（LUSC）和肺腺癌（LUAD）患者组织中比在正常组织中显著升高（P<0.05）;K-M曲线结果显示,在肺癌中RRM2高表达患者较低表达患者的总生存期显著降低（P<0.001）;人支气管上皮细胞（HBE1）RRM2表达水平显著低于NSCLC的CALU3和H1299细胞系的表达水平（P<0.05）;与对照组和si-NC组相比,si-RRM2-1组培养48 h和72 h后OD450值、凋亡率和增值迁移能力下降（P<0.05）;与对照组和pc-NC组相比,pc-RRM2-2组培养48 h和72 h后OD450值和增值迁移能力增加（P<0.05）。结论 沉默RRM2可以抑制NSCLC细胞的过表达效率、增殖活性、迁移和侵袭的能力。反之,过表达RRM2可以增强NSCLC细胞的过表达效率、增殖活性、迁移和侵袭的能力。

Abstract

Objective To explore the expression and biological behavior of Ribonucleotide Reductase Regulatory Subunit M2 (RRM2) in Non-Small Cell Lung Carcinoma (NSCLC). Methods Gene expression profiles and clinical data of NSCLC patients were extracted from The Cancer Genome Atlas (TCGA) database. The expression of RRM2 gene in normal tissues and NSCLC tissues was analyzed, and survival curves werewere plotted by utilizing the clinical data of the NSCLC patients. Quantitative Real-Time PCR (qPCR) was performed to determine RRM2 expression levels and verify silencing and overexpression efficiency in cell lines. Western Blot was applied to detect the protein expression of the RRM2 gene in cell lines and to validate the efficiency of its silencing and overexpression in cell lines. Cell Counting Kit-8 (CCK-8), flow cytometry and Transwell assays were conducted to detect cellproliferation, apoptosis, invasion, and migration abilities. Results RRM2 expression was significantly elevated in Lung Squamous Cell Carcinoma (LUSC) and Lung Adenocarcinoma (LUAD) tissues compared to normal controls (P<0.05).The results of the Kaplan-Meier (K-M) curve showed that the overall survival (OS) of patients with high RRM2 expression in lung cancer was significantly lower than that of patients with low expression (P<0.001). The expression level of RRM2 in human bronchial epithelial cells (HBE1) was significantly lower than that in NSCLC cell lines CALU3 and H1299(P<0.05). Compared with the control group and si-NC group, the OD450 values, apoptosis rates and proliferation/migration abilities in the si-RRM2-1 group decreased after 48 and 72 hours of culture (P<0.05). Conversely, the pc-RRM2-2 group exhibited increased OD450 values and proliferation/migration capacities versus control and pc-NC groups (P<0.05). Conclusion RRM2 silencing can inhibit the overexpression efficiency, proliferation, migration and invasion of NSCLC cells. Conversely, RRM2 overexpression can enhance the overexpression efficiency, proliferation activity, migration and invasion ability of NSCLC cells.

关键词

非小细胞肺癌 / RRM2 / 生物学行为

Key words

Non-small cell Lung carcinoma / RRM2 / Biological behavior

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基金资助

蚌埠医学院自然科学重点项目（2022byzd031）