miR-92a对奶山羊乳腺上皮细胞增殖及凋亡的调控分析

doi:10.11843/j.issn.0366-6964.2020.01.016

Abstract

Abstract: miRNAs are important factors in the regulation of breast tissue development and lactation function. In this study, according to existing miRNA expression profiles of breast tissues in different lactation stages of guanzhong dairy goats, miR-92a expressed differently in different lactation periods was taken as the research object to study the regulatory effect of miR-92a on the proliferation and apoptosis of goat mammary epithelial cells(GMEC),and explore its pontential regulatory genes. The expression of miR-92a in breast tissues at different lactation stages was detected by fluorescence quantitative PCR, MTT assay, EdU assay and flow cytometry, and the regulatory effect of miR-92a on the proliferation and apoptosis of GMECs and cell cycle was detected at the cell level. RNA-seq was used to analyze the differentially expressed genes in GMECs overexpressing miR-92a. qRT-PCR results showed that the expression level of miR-92a in breast tissues of dairy goats in the early lactation stage was significantly higher than that in the middle lactation stage (P<0.01), suggesting that miR-92a may play an important role in regulating lactation traits of dairy goats. After miR-92a was overexpressed in GMEC, the test results showed that compared with the NC group, the number of EdU positive cells in the miR-92a overexpressed group was significantly reduced (P<0.01), the number of cells in the S phase was significantly decreased, and the number of cells in the G1 phase was significantly increased, while the number of apoptotic cells in the miR-92a group was significantly increased (P<0.05).Transcriptome sequencing technology constructed the mRNA library of miR-92a overexpression, and found 54 genes were down-regulated, meanwhile 160 genes were up-regulated. Analysis of GO terms and KEGG pathway showed that differentially expressed genes involved in regulating various biological functions of mammary epithelial cells. The above results indicate that miR-92a promotes the apoptosis of mammary epithelial cells and inhibits cells proliferation. The sequencing results further prove that miR-92a plays an important role in the regulation of breast development and lactation performance in dairy goats.

Key words: miR-92a, dairy goat, mammary epithelial cells, RNA-Seq

CLC Number:

BAO Lijuan, LIU Yuhan, MA Yi, AN Xiaopeng, ZHANG Yue, ZHANG Meng, WANG Jiangang, DU Bin, LI Guang, CAO Binyun. The Regulatory of MiR-92a on Proliferation and Apoptosis of Dairy Goat Mammary Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(1): 137-149.

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The Regulatory of MiR-92a on Proliferation and Apoptosis of Dairy Goat Mammary Epithelial Cells

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