microRNA-96-5p靶向调控羊驼黑色素细胞中MITF基因的表达

doi:10.11843/j.issn.0366-6964.2020.007

Abstract

Abstract: The study aimed to predict and verify the key miRNA regulating the expression of microphthalmia-associated transcription factor (MITF). The online softwares were used to predict miRNAs with a targeted relationship to MITF in this study. In the melanocytes of alpaca, the expression of MITF gene after overexpression and inhibition of miR-96-5p was detected by fluorescence quantitative PCR. MITF-specific antibody staining was performed on cells in which the expression of miR-96-5p was overexpressed and inhibited by immunocytochemical techniques. The results showed that miR-96-5p was a key miRNA regulating the expression of MITF. The results of fluorescence quantitative PCR and immunocytochemical detection showed that when miR-96-5p was overexpressed, MITF mRNA level had no significant change, but the expressions of its downstream genes were extremely significantly reduced (P<0.01). MITF protein and its downstream proteins levels were extremely significantly lower than those in the blank groups (P<0.01). When miR-96-5p was inhibited, MITF mRNA level had no significant change, but the expressions of its downstream genes were extremely significantly increased (P<0.01). MITF protein and its downstream proteins levels were extremely significantly higher than those in the blank groups (P<0.01). In conclusion, miR-96-5p can target MITF gene and play a regulatory role at the post-transcriptional level of MITF, inhibiting the transcription of MITF and the expressions of its downstream genes of TYR, TYRP1 and TYRP2.

Key words: miR-96-5p, MITF, prediction of miRNAs, validation of target gene

CLC Number:

S829.9

ZHANG Lihuan, MA Yueyue, LIU Wenyan, LAN Wutao, ZHU Zhiwei. microRNA-96-5p Targets MITF Gene in Alpaca Melanocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1229-1237.

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microRNA-96-5p Targets MITF Gene in Alpaca Melanocytes

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