巨囊性病液状蛋白-15在肺癌中的表达研究

Study on the expression of gross cystic disease fluid protein-15 in lung cancer

目的探讨巨囊性病液状蛋白-15(GCDFP-15)在肺癌中的表达情况及作用。方法利用癌症基因组图谱(TCGA)数据库分析肺癌组织及癌旁组织中GCDFP-15的表达差异;不同临床分期肺癌患者的GCDFP-15的表达差异及对GCDFP-15高/低表达组的肺癌患者进行生存期比较。采用酶联免疫吸附试验(ELISA)检测肺癌患者及对照组血浆中的GCDFP-15水平;采用定量逆转录PCR(RT-qPCR)和Western blot分别检测4株肺癌细胞系与1株支气管上皮细胞系中的GCDFP-15 mRNA和蛋白表达量;ELISA实验检测以上5株细胞系培养液上清中的GCDFP-15浓度,综合分析GCDFP-15在肺癌中的表达情况。结果TCGA数据库显示在肺腺癌和鳞癌中的GCDFP-15的表达、不同临床分期肺癌患者的GCDFP-15表达、低表达组与高表达组的中位生存期比较,差异无统计学意义(P>0.05)。肺癌组患者血浆中的GCDFP-15浓度高于正常对照组,差异有统计学意义(P<0.05)。4个肺癌细胞株的GCDFP-15 mRNA均高于支气管上皮细胞,差异有统计学意义(P<0.05)。Western Blot结果显示这5株细胞均未检测到GCDFP-15蛋白,在细胞培养液上清中检测到GCDFP-15,且4株肺癌细胞系中,A549和H620培养基上清中的GCDFP-15表达均高于16HBE,差异有统计学意义(P<0.05)。结论肺癌患者血浆中的GCDFP-15蛋白高于正常对照组,4株肺癌细胞系的GCDFP-15 mRNA均高于支气管上皮细胞系,2株肺癌细胞系培养液上清中的GCDFP-15蛋白高于支气管上皮细胞系,结合TCGA数据库分析的结果,推测GCDFP-15可能是肺癌的一个肿瘤标志物。

Objective To investigate the expression and role of gross cystic disease fluid protein-15(GCDFP-15)in lung cancer.Methods The cancer genome atlas(TCGA)database was used to analyze the differences in the expression of GCDFP-15 in lung cancer tissues and adjacent tissues;the differences in the expression of GCDFP-15 in lung cancer patients with different clinical stages and then to compare the survival time of lung cancer patients in the GCDFP-15 high/low expression group.GCDFP-15 levels in plasma of lung cancer patients and control group were detected by enzyme-linked immunosorbent assay(ELISA).Quantitative reverse transcription PCR(RT-qPCR)and Western blot were used to detect the expression of GCDFP-15 mRNA and protein in 4 lung cancer cell lines and 1 bronchial epithelial cell line,respectively.The concentration of GCDFP-15 in the supernatant of culture medium of the above 5 cell lines was detected by ELISA,and the expression of GCDFP-15 in lung cancer was comprehensively analyzed.Results TCGA database showed that there were no statistically significant differences in the expression of GCDFP-15 in lung adenocarcinoma and squamous cell carcinoma,the expression of GCDFP-15 in lung cancer patients with different clinical stages,and the median survival between low expression group and high expression group(P>0.05).The plasma concentration of GCDFP-15 in lung cancer group was higher than that in normal control group,and the difference was statistically significant(P<0.05).The GCDFP-15 mRNA of 4 lung cancer cell lines was higher than that of bronchial epithelial cells,and the difference was statistically significant(P<0.05).Western Blot results showed that GCDFP-15 protein was not detected in all the 5 cell lines,and GCDFP-15 was detected in the supernatant of cell culture medium,and the expression of GCDFP-15 in the supernatant of A549 and H620 medium was high er than that of 16HBE in all the 4 lung cancer cell lines,the difference was statistically significant(P<0.05).Conclusion GCDFP-15 in plasma of lung cancer patients was higher than that of normal control group,GCDFP-15 mRNA in 4 lung cancer cell lines was significantly higher than that of bronchial epithelial cell lines,GCDFP-15 in supernatant of culture medium of 2 lung cancer cell lines was higher than that of bronchial epithelial cell lines.Combined with the results of TCGA database analysis,it is speculated that GCDFP-15 may be a tumor marker of lung cancer.