利用cDNA微阵列进行宫颈鳞癌的分子筛查(英文)

Molecular classification of cervical squamous cell carcinoma using cDNA microarrays

目的使用cDNA微阵列筛选浸润性宫颈鳞癌的淋巴结转移相关基因。方法应用包含18 432个基因的cDNA微阵列测定IB期宫颈鳞癌的全基因序列,包括已知功能的人类转录子和表达序列标签ESTs,分为正常组、淋巴转移组、无淋巴转移三组宫颈组织。为了证实不同的基因表达,选择3个基因进行了冰冻组织的RT-PCR检测和石蜡组织的免疫组化检测。结果经统计学分析,与无淋巴转移浸润性宫颈鳞癌组织比较,有淋巴转移的癌组织有677个基因大于2倍差异,其中上调494个(72.97%),下调183个(27.03%),表达序列标签EST为61个(9.01%),这些基因涉及代谢、发育、信号传导、分化、DNA结合转录和离子通道等。6倍差异基因14个,其中只有nel(chicken)like-2下调,其余为上调基因。RT-PCR和免疫组化的结果与cDNA微阵列结果一致。结论利用cDNA微阵列检测基因的表达状态可以预测宫颈鳞癌淋巴结转移和宫颈癌的预后情况。Cx43的低表达、ETV5和整合素alpha 2的高表达可能会成为评估浸润性宫颈鳞癌恶性程度的重要指标。这些分子有利于预测浸润性宫颈鳞癌的预后及其相应的分子治疗。

Objective To identify the new lymph node metastasis-associated molecular marker of invasive cervical carcinoma（ICC） by cDNA microarrays. Methods High-throughput cDNA microarrays containing 18 432 clones that correspond to either human transcripts with known function or anonymous expressed sequence tags（ESTs） were used to measure global patterns of gene expression in ICC of FIGO stage Ib compared with normal cervical tissue. The differentially expressed genes in cervical squamous with lymph node metastasis were investigated. To verify the differential genes in patient samples, several genes were selected to analyse by reverse transcription-PCR in frozen tumor tissue and by immunostaining in paraffin-embedded tissue section. Results In sample carcinoma tissue with lymph node metastases, all 677 genes were identified with differential expression, 494 of which showed increased expression（72.97 % ）, 183 decreased expression （27.03 % ）, and ESTs were 61 （9.01% ）, including genes of metabolism, development, signal transduction and differentiation. Thereinto, 14 6-fold differential genes were up-regulated except that nel （chicken）-like 2 was down-regulated. The results of RT-PCR and immunohistochemistry were in agreement with the microarray data. Conclusion The results demonstrate that gene expression profiling can be used as a predictor of lymph node metastasis and prognosis of cervical carcinoma. The prediction of cervical malignancy based on the lack of Cx43 and superabundance of ETV5 and integrin alpha 2 might serve as specific diagnostic biomarkers for ICC. These expression may be suitable for molecular classification of disease stages and prediction of treatment response in cervical carcinoma.