桥粒芯糖蛋白3基因在鼻咽癌中表达的初步研究

Expression of desmoglein 3 in nasopharyngeal carcinoma:research of 22 cases

目的从纯化鼻咽组织全基因组表达谱数据中筛选出鼻咽癌发病相关的候选靶基因桥粒芯糖蛋白3(DSG3),利用半定量逆转录(sqRT)-PCR 进一步研究 DSG3基因在纯化鼻咽部组织中反义 RNA(aRNA)中的表达。方法采用 RNA 保护技术保存组织标本(22例鼻咽癌患者及12例鼻咽部正常或存在炎性黏膜上及组织者),显微切割获得鼻咽癌组织标本中的癌细胞、鼻咽部非癌组织标本中的黏膜上皮细胞,提取每一例纯化组织标本 RNA,线性扩增获得足量 aRNA,通过 sqRT-PCR进一步研究候选靶基因 DSG3在两种不同鼻咽纯化细胞 aRNA 中的表达。结果在纯化鼻咽癌组织和鼻咽部非癌组织全基因组表达谱中,DSG3基因表达值分别为2.06±1.60和0.48±0.23,表达相差4.24倍,两组比较差异有统计学意义(df=16,t=2.145,P=0.048)。sqRT-PCR 检测纯化鼻咽癌组织及非癌组织 aRNA 中 DSG3基因的表达,分别为3.54±2.69和0.95±0.23,两组差异亦有统计学的意义(df=32,t=3.307,P=0.002)。结论利用全基因组芯片构建基因表达谱结合 sqRT-PCR 等实验研究,可筛选出与鼻咽癌发病相关的候选靶基因,DSG3基因高表达是鼻咽癌发生发展过程中的重要促瘤因素。

Objective To investigate the expression of desmoglein 3 （DSG3） , a candidate target gene in the antisense RNA （aRNA） from the purified nasopharyngeal tissues in nasopharyngeal carcinoma （NPC）. Methods Specimens of nasopharyngeal tissues were harvested from 22 NPC patients, aged 44 ± 11 （ NPC group） , and 12 normal persons or patients with nasopharyngeal infectious diseases, aged 46± 14. Microdissectlon technique was used to get homogenous tissue cells from which total RNA was isolated （control group）, aRNA was amplified from the total RNA by ＂in vitro transcription＂ （IVT）. The expression of DSG3 gene was identified using these aRNA by seml-quantitative reverse transcriptase polymerase chain reaction （sqRT-PCR）. Results The average expression level of DSG3 in the NPC group was 3. 536 ± 2. 689, significantly higher than that of the control group （0.95 ± 0.23, df= 32, t = 3. 307, P = 0. 002 ）. The expression level of DSG3 in the whole expression profiling of the NPC group was 1.06 ± 1. 60 , significantly higher than that of the control group （0.48 ± 0. 23, df = 16, t = 2. 145, P = 0. 048）. Conclusion The whole genome expression profiling detected by sqRT-PCR can be used to shift the marker genes from biopsy tissue samples. DSG3 may be a tumor candidate gene in NPC.