基于转录组测序分析SOX18在湖羊毛囊毛乳头细胞中的功能

doi:10.11843/j.issn.0366-6964.2024.06.013

Abstract

Abstract:

The purpose of this study was to preliminary investigate the function of the SOX18 gene in Hu sheep hair follicle dermal papilla cells. In this study, immunofluorescence staining was used to identify cells isolated from the hair follicles of Hu sheep. Subsequently, an overexpression vector of the SOX18 gene was conducted, and its overexpression efficacy was assessed using qRT-PCR and Western blot. The dermal papilla cells successfully identified and well-grown were divided into two groups. One group was transfected with SOX18 overexpression vector, while the other was transfected with empty plasmid. Each group had 3 replicates. The total RNA was extracted from 6 samples using the Trizol method, followed by the construction of a cDNA library. The Illumina Novaseq6000 platform was used for sequencing. Subsequently, sample correlation and differentially expressed genes were analyzed. The differentially expressed genes were categorized based on GO and annotated using the KEGG pathway analysis to identify candidate genes and pathways associated with the regulation of the SOX18 gene. To confirm the accuracy of the transcriptome data, 9 differentially expressed genes were randomly chosen for qRT-PCR analysis. The immunofluorescence results revealed dermal papilla-related genes had high expression in isolated cells, indicating a high purity of the isolated dermal papilla cells. qRT-PCR and Western blot results showed that overexpression of the SOX18 gene could increase the mRNA and protein levels of SOX18 in dermal papilla cells, indicating that the SOX18 overexpression vector could be used for subsequent experiments. Transcriptome analysis results showed a good correlation between samples. A total of 157 genes were differentially expressed between the SOX18 overexpression group and the control group, of which 103 genes were up-regulated after SOX18 overexpression and 54 genes were down-regulated. qRT-PCR analysis showed high accuracy of transcriptome data. GO functional analysis showed that some differential genes were enriched in cell progression and hair follicle development. KEGG enrichment analysis showed that some differentially expressed genes were enriched in signaling pathways related to cell proliferation and hair follicle development. GO functional analysis and KEGG enrichment analysis indicated that SOX18 played a role in the proliferation of dermal papilla cells and the development of hair follicles. In this study, transcriptome sequencing analysis found that SOX18 may affect the proliferation of dermal papilla cells and the development of hair follicles by affecting genes and signaling pathways related to cell proliferation and hair follicle development. The study results provide a theoretical basis for understanding the molecular regulatory mechanism of the SOX18 gene in Hu sheep dermal papilla cells and hair follicles.

Key words: transcriptome sequencing, SOX18, hair follicles, dermal papilla cells, Hu sheep

CLC Number:

S826.2

Mingliang HE, Xiaoyang LÜ, Yongqing JIANG, Zhenghai SONG, Yeqing WANG, Huiguo YANG, Shanhe WANG, Wei SUN. Function Analysis of SOX18 in Hu Sheep Hair Follicle Dermal Papilla Cells Based on Transcriptome Sequencing[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2409-2420.

Figures/Tables 8

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References 26

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基因 Gene	登录号 Accession number	引物序列(5′→3′) Primer sequence	产物大小/bp Product size	退火温度/℃ Annealing temperature
AHR	XM_060414533.1	F: CGAATCCTTCAAAGAGGCATAG	403	60
		R: GCTCGGTCTTCGGTATGG
ANKRD1	NM_001252178.1	F: TACAGGGAAGAAGAACGG	338	60
		R: TAGGCACATCCACAGGTT
BMP4	XM_060418703.1	F: CCGTGAGGAGCTTCCACCA	191	60
		R: GTTTATACGATGAAAGCCCTGCTCC
CFB	XM_004018929.6	F: TGTCCTTCTGGCTTCTACCC	466	60
		R: AAGGCTCTGTTCCACTCCAA
GANC	XM_060419122.1	F: AGTCTACCTATCGGGCATTA	269	60
		R: ATCTCCTTTCCCACTTGC
HUNK	XM_004003742.6	F: TTGCCAGGAAGAAATACGG	189	60
		R: GCAGGAAGTTCACGGCTC
PTGFR	NM_001009789.2	F: TTTAGAAGTCAGCAGCAC	354	60
		R: ATCAGAAATAGCAGCAAC
PTGS2	NM_001009432.1	F: CCCGAACAGGATTCTATG	481	60
		R: GCTGGTCCTCGTTCAATA
SOX9	XM_027974011.3	F: TCAAGGGCTACGACTGGACG	366	60
		R: TGGCGTTGGGCGAGATGTGC
SOX18	XM_027976914.2	F: TGTGGGCGAAGGACGAGC	253	60
		R: GCCAAGCCTGGGAGGAGGAG
GAPDH	NM_001190390.1	F: TCTCAAGGGCATTCTAGGCTAC	151	60
		R: GCCGAATTCATTGTCGTACCAG

引物名称 Primer name	引物序列(5′→3′) Primer sequence	产物大小/bp Product size	退火温度/℃ Annealing temperature
OE-SOX18	F: CTAGCGTTTAAACTTAAGCTTATGCAGAGATCGCCGCTCG	1 176	62
	R: CCACACTGGACTAGTGGATCCCTATCCAGAGATGCAGGCGCTG

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Function Analysis of SOX18 in Hu Sheep Hair Follicle Dermal Papilla Cells Based on Transcriptome Sequencing

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