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目的探讨维吾尔族妇女宫颈癌组织特异的下调表达基因谱,建立基于下调表达基因的宫颈癌早期诊断方法。方法收集维吾尔族妇女宫颈病变新鲜组织标本72例,其中宫颈鳞癌(cervical squamous cell carcino-ma,CSCC)25例,宫颈内上皮瘤变(cervical intraepithelial neoplasia,CIN)22例,正常宫颈组织(normal cervix,NC)25例。选择20例组织标本(CSCC 7例,CINⅢ6例,NC 7例),提取组织RNA,通过人类全基因组表达谱芯片及生物信息分析,筛选宫颈癌及癌前病变(CIN)特异性下调表达候选基因。通过对CSCC与正常对照(NC)组的基因差异表达谱进行比较分析,选取10种差异表达倍数较高的候选基因,利用72例宫颈病变组织RNA,对10种下调表达候选基因的表达水平进行半定量RT-PCR鉴定,确定宫颈癌及癌前病变与该基因表达水平变化的关系。结果以2倍及以上表达差异为量化标准,有下调表达候选基因112种。72例宫颈病变组织RNA的半定量RT-PCR筛查分析,发现在CSCC与正常对照组之间,FOSB、DNASE1L3、SCARA5、EGR1、ABI3BP、FOS、KLF4、RHOB、IER2和ID4等10种下调表达基因的差异均有统计学意义(P<0.05)。DNASE1L3、EGR1、FOS、KLF4和IER2等5种基因的表达水平差异在CIN与正常对照组之间有统计学意义(P<0.05),但是除了FOS基因外,其余4种基因表达在CSCC与CIN组之间无差异(P>0.05)。结论维吾尔族妇女宫颈癌发病进程可能与一系列基因的下调表达密切相关,而DNASE1L3、EGR1、FOS、KLF4和IER2等5种基因的表达水平变化可能成为宫颈癌早期预警指标。
Objective To investigate the gene expression profile of cervical cancer in Uighur women and to establish an early diagnosis method of cervical cancer based on down-regulated genes. Methods 72 cases of fresh cervical tissue specimens from Uygur women were collected, including 25 cases of cervical squamous cell carcinomas (CSCC), 22 cases of cervical intraepithelial neoplasia (CIN), normal cervix cervix, NC) 25 cases. Tissue samples of 20 cases (7 cases of CSCC, 6 cases of CIN and 7 cases of NC) were selected for tissue RNA extraction. Human genome-wide cDNA microarray and bioinformatic analysis were used to screen for down-regulated expression of cervical cancer and precancerous lesions (CIN) . By comparing and analyzing the gene differential expression profiles of CSCC and normal control (NC) group, ten candidate genes with high differential expression ratio were selected, and the expression levels of ten down-regulated expression gene candidates were determined using 72 cases of cervical lesions tissue RNA Semi-quantitative RT-PCR identification, to determine the relationship between cervical cancer and precancerous lesions and the gene expression changes. The results of 2 times and above expression differences as a quantitative standard, there are down-regulated expression of 112 candidate genes. Semi-quantitative RT-PCR analysis of RNA in 72 cases of cervical lesions showed that down-regulated expression of FOSB, DNASE1L3, SCARA5, EGR1, ABI3BP, FOS, KLF4, RHOB, IER2 and ID4 Gene differences were statistically significant (P <0.05). The differences of the expression levels of five genes (DNASE1L3, EGR1, FOS, KLF4 and IER2) between CIN and normal control group were statistically significant (P <0.05). However, except for FOS gene, the other four genes expressed in CSCC and CIN No difference between groups (P> 0.05). Conclusion The incidence of cervical cancer in Uighur women may be closely related to the down-regulation of a series of genes. The changes of the expression of 5 genes, including DNASE1L3, EGR1, FOS, KLF4 and IER2, may be the early warning indicators for cervical cancer.