应用cDNA表达芯片研究复方黄连对CNE1鼻咽癌移植瘤基因表达的影响

Effect of Coptis Chinensis compound on the gene expression in transplanted tumor tissue in nasopharyngeal carcinoma cell line of CNE1 by cDNA microarray

目的 :研究复方黄连对鼻咽癌裸鼠移植瘤基因表达的影响。方法 :培养的鼻咽癌细胞系CNE1细胞接种Balb/C裸鼠 ,待移植瘤形成后灌喂复方黄连 ,然后从移植瘤组织提取RNA ,经逆转录标记后的cDNA作为探针 ,与高通量cDNA表达芯片杂交并用激光共聚焦扫描仪分析基因表达。通过逆转录 聚合酶链式反应 (RT PCR)半定量技术进一步分析差异表达基因的表达。结果 :复方黄连处理移植瘤裸鼠 30d后 ,移植瘤明显减小 ,其抑瘤率为 2 9.5 %。同时 ,移植瘤组织基因表达发生显著变化。用该复方处理的CNE1鼻咽癌移植瘤组织有 147条基因表达发生改变 ,包括 10 2条下调表达的基因和 45条上调表达的基因。RT PCR半定量技术分析证实有 3条基因为真正差异表达 ,它们分别是MAD3,H731和CHK1基因。其中 ,MAD3和H731基因表达上调 ,CHK1基因表达下调。结论 :复方黄连可抑制CNE1鼻咽癌裸鼠移植瘤的生长并可影响其基因的表达 ,它对CNE1鼻咽癌移植瘤的抑制作用与其调节移植瘤基因表达密切相关。

Objective To investigate the effect of Coptis Chinensis compound on the gene expression of transplanted tumor in nasopharyngeal carcinoma cell line of CNE1. Methods The cells were injected into Balb/C nude mice, and after the transplanted tumor was established , the nude mice with tumors were treated with Coptis chinensis compound. The nude mice were then killed, tumor tissues were isolated and the total RNA was extracted. Hybridization with a cDNA microarray was conducted using a probe of fluorescent-labeled cDNA mixture that was reversely transcribed from the RNA. The gene expression level was analyzed after scanning the fluorescent intensity by a laser scanner, and further confirmed by RT-PCR. Results After using Coptis Chinensis compound for 30 days, the size of transplanted tumors were remarkably reduced, and the inhibition rate of tumor growth was 29.5%. In the meantime, the expression of 147 genes in the tissue of implanted tumor treated with the compound altered:the expression of 102 genes was down-regulated, and that of 45 genes was up-regulated. Furthermore, the differential expression of 3 genes was verified by RT-PCR, MAD3 and H731 genes were up-regulated, and that of CHK1 was down-regulated. Conclusion Coptis Chinensis compound can affect the gene expression in the tissue of implanted tumor, which implies that the inhibition of Coptis Chinensis compound on the growth of the implanted tumor with CNE1 cells in nude mice might be associated with the control of gene expression.