MicroRNA-216b inhibits heat stress-induced cell apoptosis by targeting Fas in bovine mammary epithelial cells
MicroRNA-216b inhibits heat stress-induced cell apoptosis by targeting Fas in bovine mammary epithelial cells
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DOI：
10.1007/s12192-018-0899-9
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发表时间：
2018-09-01
影响因子：
3.8
通讯作者：
Lai, Songjia
中科院分区：
文献类型：
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作者：
Cai, Mingcheng;Hu, Yongsong;Lai, Songjia
夏季热应激影响泌乳奶牛产奶量和乳品质。牛乳腺上皮细胞（bMEC）在乳汁分泌中起关键作用，而microRNAs（miRNAs）调控bMEC的多种功能。已有研究证实miR-216 b通过抑制靶基因来调控多种肿瘤细胞的凋亡。因此，我们的目的是探索miR-216 b在热应激诱导的bMEC细胞凋亡中的潜在参与。首先，我们建立了热应激模型，发现热应激条件下bMEC的凋亡率显著增加。miR-216 b、Bax mRNA和caspase-3 mRNA表达上调。而Bcl-2 mRNA表达则呈差异性下调。过表达miR-216 b可显著下调caspase-3和Bax的mRNA和蛋白表达，上调Bcl-2的mRNA和蛋白表达。抑制miR-216 b可增加caspase-3和Bax的活性，抑制Bcl-2的表达。通过生物信息学分析和双荧光素酶报告基因分析，证实Fas是miR-216 b的靶基因。转染miRNA模拟物和抑制剂后，Fas活性分别显著抑制和增强。最后，通过小干扰RNA（siRNA）抑制Fas也抑制热应激诱导的细胞凋亡。综上所述，我们的结果表明，miR-216 b通过靶向Fas在bMEC中发挥热应激下的抗凋亡作用。
Heat stress affects milk yield and quality in lactating dairy cows in summer. Bovine mammary epithelial cells (bMECs) play a key role in milk secretion, and microRNAs (miRNAs) regulate numerous functions of bMEC. Previous reports have verified that miR-216b regulated cell apoptosis through repressing target genes in several cancer cells. So, our purpose was to explore the potential involvement of miR-216b in heat stress-induced cell apoptosis in bMECs. Firstly, the heat stress model was constructed and we found that apoptotic rates of bMECs significantly increased under heat stress. The expression of miR-216b, Bax mRNA, and caspase-3 mRNA was upregulated. However, Bcl-2 mRNA level was detected to differentially downregulated. Overexpression of miR-216b remarkably downregulated the expression of caspase-3 and Bax mRNA and protein, and the mRNA and protein level of Bcl-2 was increased. Inhibition of miR-216b increased the activity of caspase-3 and Bax, and the level of Bcl-2 was inhibited. Moreover, Fas was identified as a target gene of miR-216b through bioinformatic analysis and dual-luciferase reporter assay. Fas activity was significantly inhibited and enhanced respectively after transfecting miRNA mimics and inhibitor. Finally, inhibition of Fas via the small interfering RNA (siRNA) also inhibited cell apoptosis induced by heat stress. Taken together, our results indicated that miR-216b exerted as an anti-apoptotic effect under heat stress in bMECs by targeting Fas.