摘要
目的利用Agilent全基因组寡核苷酸基因表达谱芯片探寻与卵巢癌转移和耐药相关的基因表达序列,初步探讨卵巢癌转移耐药相关的分子机制。方法通过全基因组基因芯片检测高转移性及耐药性的上皮性卵巢癌细胞系RMG-1-H、COC1/DDP和HO8910/PM与其各自对应的RMG-1-C、COC1和HO8910细胞系基因表达序列,探寻差异表达的基因并对其进行基因本体论分析、通路分析及交互作用网络分析,并用实时PCR和免疫组织化学方法验证芯片结果。结果有49个基因发生2倍以上的表达差异,主要涉及基因表达、生物聚合物的合成等方面,交互网络图预测出对相互作用发挥连接作用的21个基因。采用实时聚合酶链反应及免疫组织化学方法对差异表达基因GCET2、CFTR、FOXP1和GARS在细胞和组织中进行基因及蛋白水平验证,结果与芯片结果相一致。结论与卵巢癌转移和化疗耐药相关的基因表达谱序列的变化可以为卵巢癌转移和化疗耐药的相关分子机制研究提供理论基础。
Objective To investigate the genes associated with higher ability of metastasis and chemotherapic resistance in epithelial ovarian carcinoma（ EOC） by using Agilent whole genome oligonucleotide gene chip,with an attempt to further investigate the molecular mechanism of metastasis and chemotherapic resistance of EOC. Methods Oligonucleotide microarrays were used to determine whether gene expression profile might differentiate EOC cell lines（ RMG-1-C,COC1 and HO8910） from their sub-lines（ RMG-1-H,COC1 / DDP and HO8910 / PM） with higher ability of metastasis and chemotherapic resistance. Quantitative real-time polymerase chain reaction and immunohistochemical staining validated the microarray results. Results Gene expression pro-file identified 49 differentially expressed genes that showed≥2. 0 fold change. All these differentially expressed genes were involved mainly in gene expression and biopolymer biosynthesis. Interaction network analysis predicted21 genes participating in the regulatory connection. Highly differential expression of GCET2, CFTR, FOXP1 and GARS genes was validated by quantitative realtime polymerase chain reaction in all cell line samples,and the results were consistent with microarray findings. Conclusion The change in the metastasis and chemotherapic resistance-associated gene expression profiles may provide a theoretical basis for studies on the molecular mechanisms of metastasis and chemotherapic resistance in EOC.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2015年第6期662-673,共12页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(81072118
81172491
81101527)
高等学校博士科学点专项科研基金(20112104110016
20112104120019)
中国医科大学盛京医院自由研究者计划项目(200807)~~
关键词
基因表达序列
转移相关基因
化疗耐药相关基因
微阵列
上皮性卵巢癌
FOXP1
gene expression profile
metastasis-associated gene
drug-resistance-associated genes
microarray
epithelial ovarian carcinoma
FOXP1
