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摘要:
目的 研究葡萄糖胺基转移酶3(glycosyltransferase enzyme 3, GCNT3)在非小细胞肺癌(non-small cell lung cancer, NSCLC)组织及对应正常组织中的表达情况, 探讨其表达水平与NSCLC患者临床病理特征、总生存期(overall survival, OS)和无进展生存期(progression-free survival, PFS)的关系。 方法 分别应用实时定量反转录链式反应(qRT-PCR)和蛋白质印记技术(Western blot, WB)检测GCNT3在2017年3月至2017年7月天津医科大学肿瘤医院20例NSCLC患者癌组织及对应的正常组织中的表达情况; 此外, 收集2010年1月至2014年12月本院164例NSCLC患者的石蜡组织标本, 通过免疫组织化学技术(immunohistochemistry, IHC)评估样本中GCNT3表达, 分析GCNT3表达水平与临床病理学特征之间的关系, 探究GCNT3表达与NSCLC患者OS及PFS的关系, 通过细胞功能实验研究GCNT3对NSCLC细胞增殖能力、侵袭能力和迁移能力的影响。 结果 qRT-PCR与WB结果均显示GCNT3在NSCLC癌组织中的表达水平明显高于正常组织。IHC结果显示GCNT3表达水平与NSCLC患者的性别、吸烟史、组织学类型、病理分期和淋巴结转移相关; Kaplan-Meier分析显示GCNT3高表达NSCLC患者OS和PFS均差于GCNT3低表达患者(P < 0.05)。多因素Cox比例风险回归模型分析显示, GCNT3表达是NSCLC患者预后的独立因素(P < 0.05)。抑制GCNT3表达后, NSCLC细胞的增殖能力、侵袭能力和迁移能力明显减弱(P < 0.05)。 结论 GCNT3在NSCLC癌组织中高表达, GCNT3高表达的NSCLC患者OS和PFS较差, GCNT3有望成为评估NSCLC患者预后的标志物。 Abstract:Objective To investigate the expression of glycosyltransferase enzyme 3 (GCNT3) in non-small cell lung cancer (NSCLC) tissues and corresponding normal tissues, and to further explore the relationship between GCNT3 expression and clinicopathological features, overall survival (OS), and progression-free survival (PFS) in patients with NSCLC. Methods In this study, we used quantitative real-time polymerase chain reaction and Western blot to assess the mRNA and protein expression of GCNT3 in paired NSCLC and non-tumor tissues. In addition, 164 NSCLC patients were estimated for GCNT3 expression by immunohistochemistry, and the correlation between GCNT3 expression and clinicopathological features was evaluated. Further, the effects of GCNT3 on the proliferation, invasion, and migration abilities of NSCLC cells were studied. Results The mRNA and protein expression levels of GCNT3 in NSCLC tissues were both significantly higher than those in the corresponding non-tumor tissues. Among the 164 patients with NSCLC, high GCNT3 expression was associated with gender, smoking, histology, pathological stage, and lymph node metastasis. Kaplan-Meier analysis displayed significant differences in OS and PFS among the groups exhibiting differences in GCNT3 expression (P < 0.05). The NSCLC patients with increased GCNT3 expression showed poor OS and PFS. A multivariate analysis demonstrated that GCNT3 expression was as an independent prognostic factor for NSCLC (P < 0.05). Cell function experiments showed that the proliferation, invasion, and migration abilities of NSCLC cells were significantly attenuated after inhibition of GCNT3 expression (P < 0.05). Conclusions High expression of GCNT3 was associated with unfavorable OS and PFS in patients with NSCLC; GCNT3 might, therefore, act as a prognostic biomarker for NSCLC. Key words glycosyltransferase enzyme 3 (GCNT3), non-small cell lung cancer (NSCLC), immunohistochemistry, prognosis. -
图 2 免疫组织化学法检测NSCLC癌组织中GCNT3表达
A、B、C、D分别为GCNT3在正常肺组织及NSCLC组织中阴性(鳞癌)、弱阳性(腺癌)、强阳性(腺癌)表达(IHC×200);E、F、G、H分别为GCNT3在正常肺组织及NSCLC组织中阴性(鳞癌)、弱阳性(腺癌)、强阳性(鳞癌)表达(IHC×400)
图 4 转染shRNA后GCNT3的表达情况及细胞增殖、细胞迁移实验结果(shNC:未转染组; shRNA:转染组)
►A:转染后GCNT3在A549中的表达; B:抑制GCNT3表达对A549细胞增殖率的影响; C:抑制GCNT3表达对A549细胞侵袭力的影响; D:抑制GCNT3表达对A549细胞迁移能力的影响
表 1 GCNT3表达与NSCLC临床病理特征的关系
表 2 影响NSCLC患者预后的单因素分析
表 3 影响NSCLC患者预后的多因素分析
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