lnc9141-a and -b Play a Different Role in Bovine Myoblast Proliferation, Apoptosis, and Differentiation

Meng Zhang; Bo Li; Jian Wang; Sihuan Zhang; Hui Li; Lin Ma; Wei Guo; Chuzhao Lei; Hong Chen; Xianyong Lan

doi:10.1016/j.omtn.2019.09.013

lnc9141-a and -b Play a Different Role in Bovine Myoblast Proliferation, Apoptosis, and Differentiation

Mol Ther Nucleic Acids. 2019 Dec 6:18:554-566. doi: 10.1016/j.omtn.2019.09.013. Epub 2019 Sep 23.

Authors

Meng Zhang¹, Bo Li¹, Jian Wang¹, Sihuan Zhang¹, Hui Li¹, Lin Ma¹, Wei Guo², Chuzhao Lei¹, Hong Chen¹, Xianyong Lan³

Affiliations

¹ Shaanxi Key Laboratory of Animal Genetics, Breeding & Reproduction, Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.
² Animal Science Wyoming INBRE Program, University of Wyoming, University Avenue, Laramie, WY 82071, USA.
³ Shaanxi Key Laboratory of Animal Genetics, Breeding & Reproduction, Shaanxi Key Laboratory of Molecular Biology for Agriculture, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China. Electronic address: lan342@126.com.

Abstract

Previously, our transcriptome sequencing revealed that lnc9141 was differentially expressed in muscles of fetal bovine, calf, and adult bovine, which is considered to provide the basis for raising the muscle mass. In this study, we identified lnc9141 characters. lnc9141 has different transcription start sites and 3' alternative splicing sites of exon 1, producing lnc9141-a and lnc9141-b transcripts that were highly expressed in the heart and lung. Moreover, neither lnc9141-a nor lnc9141-b had the ability to encode proteins. The functions of lnc9141-a and lnc9141-b were explored by cell cycle, 5-ethynyl-2'-deoxyuridine (EdU), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results showed that lnc9141-a or lnc9141-b overexpression decreased the number of myoblasts in the S phase and increased the proportion of cells in the G₀/G₁ phase. Furthermore, overexpressing lnc9141-a and lnc9141-b respectively downregulated the expression of Cyclin D1. However, lnc9141-a or lnc9141-b interference was found to increase the number of S-phase myoblasts, and upregulate Cyclin D1 and Cyclin E expression. Through Annexin V-FITC/propidium iodide (PI) double staining and the expression of apoptosis marker genes (Bax, Bcl2, and Caspase-3), it was found that lnc9141-b could regulate the expression of Bax gene. Meantime, high expression of lnc9141-b could decrease MyHC expression. In addition, the intergenic region between lnc9141 and IRX5 was 2.3 kb, with a head-to-head orientation. The study also revealed the core regions of the lnc9141 and IRX5 promoter. Our study demonstrated that both lnc9141-a and -b expression inhibited bovine myoblast proliferation. However, lnc9141-b regulated Bax and MyHC expression. The regulatory mechanism of lnc9141-a and lnc9141-b needs to be further explored.

Keywords: bovine; different transcripts; lnc9141; long non-coding RNA; myoblasts.