Differentiaiiy Of MicroRNAs Expression Profiling Of Nasopharyngeal Carcinoma

[Objective] To obtain the differential miRNAs expression profile between nasopharyngeal carcinoma tissues and normal nasopharyngeal tissues by miRNA microarray technique, verify the reliability of the results by qRT-PCR technique, analyze the the expression level of difference miRNAs in tissuses of nasopharyngeal carcinoma and nasopharyngeal cells,analyze the signal pathways that relate to the differential expression profile of miRNAs by bioinformatics.[Methods] Nasopharyngeal carcinoma tissues and Mucosa of chronic rhinitis tissues were collected and RNAs were extracted to detect the total RNA purity. The differential miRNAs expression were obtained in nasopharyngeal carcinoma tissues and Mucosa of chronic rhinitis tissue by miRNA microarray. We verified the expression level of miR-205 and miR-34c-3p in nasopharyngeal carcinoma tissues, Mucosa of chronic rhinitis tissues, Immortalized human cells NP69, and nasopharyngeal carcinoma cell line 6-10 B, 5-8F, CNE1, CNE2 by qRT-PCR technique. and The results was compared with gene miRNA microarray. We analyzed the signal pathways that relate to the differential expression profile of mi RNAs by bioinformatics.[Results] 1. The differential miRNAs expression profile was obtained by miRNA microarray. It showed 133 differential mi RNAs, which of were 31 miRNAs downregulation miRNAs and 102 miRNAs upregulation in the nasopharyngeal tissues. Among of them, miR-205 was the most significantly upregulated and mi R-34c-3p was the most significantly downregulated. 2.The expression level of mi R-34c-3p and mi R-205 in nasopharyngeal carcinoma tissues were 5.458 and-3.098 times more than Mucosa of chronic rhinitis, and the results of qRT-PCR were consistent with the results of miRNA microarray. 3. The mi R-205 expression levels in different differentiation and metastasis of nasopharyngeal carcinoma cells were higher than those in NP69 cells, and the differences were statistically significant(p<0.01).The expression level of miR-205 in highly metastatic cell line 5-8F was higher than that in low metastatic cell line 6-10 B. The expression level of miR-205 inpoorly differentiated cell line CNE2 was higher than that in high differentiated cell line CNE1( p < 0.01). 4. The expression level of miR-34c-3p in different differentiation and metastasis of nasopharyngeal carcinoma was lower than that in NP69 cells, and the differences were statistically significant(p <0.01). The expression level of mi R-34c-3p was no significant difference between highly metastatic cell lines 5-8F and low metastatic cell line 6-10B(p> 0.05). However, the expression level of miR-34c-3p in poorly differentiated cell line CNE2 was higher than that inhigh cell lines CNE1(p <0.01). 5. Bioinformatics analysis found that signaling pathways relating to target genes regulated by differential miRNAs mainly included MAPK,Focal adhesion,Insulin,Neurotrophin,ErbB,Axon guidance. Among of them, MAPK signaling pathway is closely related to the development of NPC.[Conclusion] 1. The differential miRNAs expression profile between nasopharyngeal carcinoma tissues and normal nasopharyngeal tissues was obtained by miRNA microarray technique, that 31 miRNAs were downregulated, 102 miRNAs were upregulated. 2. The expression levels of miR-205 and miR-34c-3p in nasopharyngeal carcinoma tissues and cells by qRT-PCR were consistent with the results obtained by the miRNA microarray,and the miRNA microarray results are reliable. 3. miR-205 upregulation and miR-34c-3p downregulation may play an important role in the development of nasopharyngeal carcinoma. 4. The high expression of miR-205 was associated with differentiation and metastasis of nasopharyngeal carcinoma.The low expression of miR-34c-3p was associated with differentiationand was not associated to the metastasis of nasopharyngeal carcinoma.