Screening differentially expressed genes from cyclin B1 downregulated by high-throughput gene chip

Xian-He Xie, Wan-Zun Lin, Wei-Li Zheng
2018 Journal of Hainan Medical University  
Objective: To study differentially expressed genes by silencing cyclin B1, and to sift out autophagy-related genes. Methods: Double thymidine deoxyribonucleoside blocking was used to synchronize nasopharyngeal carcinoma cell (CNE-2) to S phase, then flow cytometry was applied to test transfection efficiency. The mRNA and protein expression level of cyclin B1 was assessed by q-PCR and western blot, respectively. Differentially expressed genes were screened by high-throughput gene chip. Results:
more » ... ene chip. Results: Double thymidine deoxyribonucleoside (2.5 mmol/L) blocking was used to synchronize the cell cycle to S phase. The transfection efficiency of CNE-2 cells was 85.6%. Compared with negative group,cyclin B1-siRNA treated group significantly down-regulated mRNA expression of cyclin B1 (80%) and protein level (75.3%). Totally, 2 408 differentially expressed genes were found in CNE-2, including 1 245 up-regulated genes and 1 163 down-regulated genes. Moreover, PTEN, an autophagyrelated gene, was preliminarily sifted out. Conclusions: Cyclin B1-siRNA significantly downregulated the expression of cyclin B1 and yielded a total of 2 408 differentially expressed genes, including PETN (an autophagy-related gene).
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