LncRNA-UCA1、HMGB1/RAGE在非小细胞肺癌中的表达和意义

Expression and significance of lncRNA-UCA1，HMGB1/RAGE in non-small cell lung cancer

摘要:

目的：明确长链非编码RNA-尿路上皮癌胚抗原1（long non-coding RNA-human urothelial carcinoma associated 1，LncRNA-UCA）、高迁移率族蛋白 B1（high mobility group box-B1，HMGB1）、晚期糖基化终末产物受体（receptor for advanced glycation end-products，RAGE）、核因子-κB（nuclear factor kappa-B，NF-κB）p65在人体非小细胞肺癌（non-small cell lung cancer，NSCLC）组织及癌旁正常组织中的表达，探究其与NSCLC临床特点、病理类型等的关系。方法：选取2018年5月至2018年8月青岛大学附属医院收治的40例NSCLC患者，采集肺癌组织及癌旁正常组织标本各40例。应用免疫组化法检测10例患者肺癌组织及正常组织中的HMGB1、RAGE定位及表达量。应用Western blot检测12例NSCLC患者的肺癌组织及正常组织中HMGB1、RAGE、NF-κB p65蛋白的表达情况。采用qRT-PCR测定40例NSCLC患者肺癌组织及正常组织中lncRNA-UCA1、HMGB1、RAGE、NF-κB p65的mRNA表达量。结果：①免疫组化及Western blot检测显示，HMGB1（1 459.72±366.67）、NF-κB p65（479.52±265.07）在NSCLC中的表达量较正常组织（847.60±313.66、135.53±109.61）明显升高（t=5.470，P=0.000；t=4.063，P=0.002），RAGE（127.49±43.91）在 NSCLC中的表达较正常组织（257.61±64.84）明显降低（t=-11.461，P=0.000）。②qRT-PCR 检测显示，LncRNA-UCA1（4.85±2.31）、HMGB1（4.64±2.10）、NF-κB p65（4.22±2.10）在NSCLC中的表达较正常组织（1.00±0.00、1.00±0.00、1.00±0.00）明显升高（t=10.521，P=0.000；t=10.949，P=0.000；t=8.273，P=0.000），RAGE（0.41±0.19）在NSCLC中的表达较正常组织（1.00±0.00）降低（t=-19.754，P=0.000）。③LncRNA-UCA1、HMGB1、NF-κB p65的表达量在腺癌组织（2.93±1.40、3.06±1.36、2.17±0.87）高于鳞癌组织（6.77±2.22、6.16±2.20、5.96±2.90）（t=3.722，P=0.001；t=3.114，P=0.004；t=3.348，P=0.002），大于3 cm肿瘤组织（6.70±2.14、6.28±1.80、6.07±2.17）高于不足3 cm肿瘤组织（3.11±1.40、3.21±1.43、2.28±0.86）（t=-5.367，P=0.000；t=-5.181，P=0.000；t=-5.623，P=0.000），有淋巴结转移癌组织（6.38±2.25、6.04±2.25、5.72±2.81）高于无淋巴结转移癌组织（2.70±1.30、3.23±1.38、2.24±0.92）（t=2.377，P=0.023；t=2.404，P=0.021；t=2.410，P=0.021），而RAGE的表达量在腺癌组织（0.54±0.18）低于鳞癌组织（0.28±0.12）（t=-3.049，P=0.004），大于3 cm肿瘤组织（0.28±0.11）低于不足3 cm肿瘤组织（0.54±0.18）（t=3.870，P=0.000），有淋巴结转移癌组织（0.27±0.11）低于无淋巴结转移癌组织（0.52±0.18）（t=-2.541，P=0.015）。④Pearson相关性分析表明，在NSCLC患者的癌组织中，HMGB1与lncRNA-UCA1、NF-κB p65表达呈正相关（r=0.754，P=0.000；r=0.704，P=0.000），RAGE与HMGB1、NF-κB p65表达呈负相关（r=-0.689，P=0.000；r=0.704，P=0.000）。结论：lncRNA-UCA1的高表达与NSCLC的发生发展有关；lncRNA-UCA1可能在影响HMGB1/RAGE表达及NF-κB通路中发挥作用。

Abstract:

Objective：To clarify the expression of long non-coding RNA-Human urothelial carcinoma associated 1（LncRNA-UCA），high mobility group box-B1（HMGB1），receptor for advanced glycation end-products（RAGE） and nuclear factor kappa-B（NF-κB） p65 in human non-small cell lung cancer（NSCLC） tissues and normal tissues，and to explore its relationship with the clinical fea－tures and pathological types of NSCLC. Methods：Forty patients with NSCLC admitted to our hospital from May 2018 to August 2018 were enrolled. Cancer tissues and adjacent normal tissues of 40 patients were respectively collected. The expression of HMGB1 and RAGE in cancer tissues and normal tissues 10 patients were detected by immunohistochemistry. The expression of HMGB1，RAGE，NF-κB p65 protein in cancer tissues and normal tissues of 12 patients was detected by Western blot. The mRNA expression of LncRNA-UCA1，HMGB1，RAGE and NF-κB p65 in cancer tissues and normal tissues of 40 patients were determined by quantitative real-time polymerase chain reaction（qRT-PCR）. Results：Immunohistochemistry and Western blot analysis showed that expression of HMGB1（1459.72±366.67） and NF-κB p65（479.52±265.07） in the NSCLC tissues was significantly higher than that in the adjacent normal tissues（847.60±313.66，135.53±109.61）（t=5.470，P=0.000；t=4.063，P=0.002），while expression of RAGE（127.49±43.91） in the NSCLC tissues was significantly lower than that in the adjacent normal tissues（257.61±64.84）（t=-11.461，P=0.000）. The qRT-PCR detection showed that expression of LncRNA-UCA1（4.85±2.31），HMGB1（4.64±2.10） and NF-κB p65（4.22±2.10） in the NSCLC tissues was significantly higher than that in the adjacent normal tissues（1.00±0.00），while expression of RAGE（0.41±0.19） in the NSCLC tissues was significantly lower than that in the adjacent normal tissues（1.00±0.00）（t=-19.754，P=0.000）. The expression of LncRNA-UCA1，HMGB1 and NF-κB p65 in the adenocarcinoma（2.93±1.40，3.06±1.36，2.17±0.87） was higher than that in the squamous cell carcinoma（6.77±2.22，6.16±2.20，5.96±2.90）（t=3.722，P=0.001；t=3.114，P=0.004；t=3.348，P=0.002），in the cancer tissue more than 3 cm（6.70±2.14，6.28±1.80，6.07±2.17） was higher than that in tissues less than 3 cm（3.11±1.40，3.21±1.43，2.28±0.86）（t=-5.367，P=0.000；t=-5.181，P=0.000；t=-5.623，P=0.000），and in cancer tissues with lymph node metastasis（6.38±2.25，6.04±2.25，5.72±2.81） was higher than that in cancer tissues without lymph node metas－tasis（2.70±1.30，3.23±1.38，2.24±0.92）（t=2.377，P=0.023；t=2.404，P=0.021；t=2.410，P=0.021）. The expression of RAGE in adenocarcinoma tissues（0.54±0.18） was lower than that in squamous cell carcinoma tissues（0.28±0.12）（t=-3.049，P=0.004），in tumor tissues more than 3 cm（0.28±0.11） was lower than that in tumor tissues less than 3 cm（0.54±0.18）（t=3.870，P=0.000），and in cancer tissues with lymph node metastasis（0.27±0.11） was lower than that in cancer tissues without lymph node metastasis（0.52±0.18）（t=-2.541，P=0.015）. Pearson correlation analysis showed that in cancer tissues of patients with NSCLC，HMGB1 was positively correlated with expression of LncRNA-UCA1 and NF-κB p65（r=0.754，P=0.000；r=0.704，P=0.000），while RAGE was negatively correlated with expression of HMGB1 and NF-κB p65（r=-0.689，P=0.000；r=0.704，P=0.000）. Conclusion：The high expression of LncRNA-UCA1 is related to the occurrence and development of NSCLC；LncRNA-UCA1 may play a role in influencing HMGB1/RAGE expression and NF-κB pathway.