牦牛Smad4基因mRNA组织表达谱及其miRNAs初步研究

Preliminary Studies on Tissue Expression Profiles of Smad4 mRNA and Its miRNAs in Yak

本研究旨在了解牦牛Smad4基因mRNA组织表达谱,并探索研究可能靶定Smad4基因的miRNAs。利用RT-PCR技术对牦牛Smad4基因的mRNA在下丘脑、垂体、心、肝、脾、肺、肾、股二头肌、下颌淋巴结、卵巢、输卵管、子宫12种组织中的表达谱进行分析,并运用TargentScan和miRBase软件预测可能靶定牛Smad4基因的miRNAs,并分析其在牦牛上的组织表达谱。结果表明,Smad4基因mRNA在所检测的牦牛12种组织中均有表达,尤其是在生殖系统的卵巢、输卵管、子宫组织中广泛表达;预测到可能靶定牦牛Smad4基因的miRNA共20个,从中选择6个进行表达谱分析发现,bta-miR-106a和bta-miR-377在所检测的12种组织中均有表达,bta-miR-146a、bta-miR-224和bta-miR-1434-5p除了在输卵管中没有表达外,在其他组织均有表达,bta-miR-212只在卵巢中有表达。综上表明,Smad4基因在牦牛各组织中可能发挥重要作用,bta-miR-212可能仅在卵巢组织作用于靶定Smad4基因,并参与牦牛繁殖生理过程调控,其机制有待进一步研究证实。

The aim of this study was to investigate the mRNA expression profile of Smad 4 gene and to explore the possible miRNAs that target to Smad 4 gene in yak. The reverse transcription PCR（RT PCR） was used to detect the expression of Smad 4 gene mRNA in the 12 yak tissues including hypothalamus, pituitary, liver, heart, spleen, lung, kidney, biceps femoris, lymph nodes, ovary,oviduct and uterus. The possible miRNAs that target to Smad 4 gene were predicted by TargentScan and miRBase softwares, and their tissue expression profiles in yak were also ana- lyzed by RT-PCR. The results showed that Smad 4 mRNA was expressed in the 12 yak tissues, especially widely expressed in the ovary, oviduct and uterus of the reproductive system. It was predicted that there were 20 miRNAs that could target to the Smad 4 gene of yak, and 6 were selected for expression analysis. The results indicated that bta-miR-106a and bta-miR-377 were expressed in the 12 detected tissues, and bta-miR-146a, bta-miR-224 and bta-miR-1434-5p were expressed in other tissues except for the oviduct of yak, whereas bta-miR-212 was expressed only in the ovaries. The results suggest that Smad 4 gene may play important roles in the various tissues of yak, and bta-miR-212 may be a miRNA targeting to Smad 4 gene only in the ovary of yak, which possibly participates in reproductive physiological process in yak, needing further studies to clarify the mechanism.