Application of chi-miR-1388-3p gene in melatonin-mediated regulation of cashmere growth in cashmere goats

Abstract

This invention relates to the field of biotechnology, and more particularly to the application of the chi-miR-1388-3p gene in the melatonin-mediated regulation of cashmere growth in cashmere goats. This invention discovers a microRNA (chi-miR-1388-3p) associated with the growth cycle of cashmere goat hair follicles, which can regulate the proliferation, cell cycle, or apoptosis of hair papilla cells in cashmere goat skin, thereby regulating cashmere goat hair follicle growth and cashmere growth. Furthermore, this invention is the first to discover that melatonin-mediated interference with chi-miR-1388-3p promotes the proliferation of hair papilla cells in cashmere goat skin, regulates the cell cycle, and inhibits apoptosis, providing valuable insights into the regulatory role of melatonin-mediated miRNAs in cashmere goat hair follicle growth.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to the application of the chi-miR-1388-3p gene in the melatonin-mediated regulation of cashmere growth in cashmere goats. Background Technology

[0002] The Inner Mongolian cashmere goat is a popular sheep breed, renowned worldwide for its exceptional fine fiber production. Cashmere, often referred to as "soft gold," is a high-quality textile material characterized by its long, fine fibers, high moisture absorption, and excellent warmth retention. Cashmere goats produce both wool and cashmere; the cashmere originates from secondary hair follicles (SHF) beneath the epidermis, while the coarse wool comes from primary hair follicles (Geng et al., 2013; Qiao et al., 2016; Zheng et al., 2019). Melatonin (MT) is an indole-like neurohormone secreted primarily at night by the pineal gland in a circadian rhythm (Claustrat et al., 2015). Notably, a similar cyclical relationship exists between serum melatonin levels and hair follicle growth, influenced by light-dark cycles, seasonal patterns, environmental factors, and reproductive rhythms (Fischer et al., 2009). Numerous studies have shown that melatonin can promote the development of secondary hair follicles, thereby promoting cashmere growth and increasing cashmere production (Duan et al., 2017; Yang et al., 2021).

[0003] MicroRNAs (miRNAs) are short, endogenous non-coding RNA molecules found in eukaryotic cells, typically 18 to 25 nucleotides in length. They interact with the 3′UTR of target messenger RNA (mRNA), leading to translational repression or mRNA degradation (Bartel et al., 2009; Ambros et al., 2011). miRNAs play a crucial role in regulating the development and growth cycle of hair follicles in fur-bearing animals by modulating transcription factors and signaling cascades. Numerous studies have shown that miRNAs are involved in almost all fundamental biological and physiological processes, including hair follicle development and cycle. For example, miRNA-203 regulates the development of wool goat hair follicles by downregulating DDOST and NAE1 (Ma et al., 2021). miR-125b, miR-181a, and miR-200b are associated with wool curl and affect hair follicle growth directly or indirectly by targeting related genes (Lv et al., 2021). Furthermore, Oku-miR-205-5p regulates rabbit hair follicle growth and increases DPC apoptosis by altering the expression of genes and proteins involved in the PI3K / Akt, Wnt, Notch, and BMP signaling pathways, ultimately changing Rex rabbit hair density (Liu et al., 2020). Therefore, miRNAs play a crucial role in the regulation of hair follicle growth and development. Summary of the Invention

[0004] In a first aspect, the present invention provides the application of the chi-miR-1388-3p gene, or the miRNA encoded by said gene, or biological material containing said gene or said miRNA in regulating the proliferation, cell cycle, or apoptosis of hair papilla cells in cashmere goat skin; the nucleotide sequence of said chi-miR-1388-3p gene is shown in any of the following:

[0005] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0006] (2) The nucleotide sequence complementary to (1);

[0007] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0008] In a second aspect, the present invention provides the application of the chi-miR-1388-3p gene, or the miRNA encoded by said gene, or biological material containing said gene or said miRNA in regulating the growth of goat hair follicles; the nucleotide sequence of said chi-miR-1388-3p gene is shown in any of the following:

[0009] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0010] (2) The nucleotide sequence complementary to (1);

[0011] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0012] Thirdly, the present invention provides the application of the chi-miR-1388-3p gene, or the miRNA encoded by said gene, or biological material containing said gene or said miRNA in regulating cashmere growth in cashmere goats; the nucleotide sequence of said chi-miR-1388-3p gene is shown in any of the following:

[0013] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0014] (2) The nucleotide sequence complementary to (1);

[0015] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0016] Fourthly, the present invention provides the application of the chi-miR-1388-3p gene, or the miRNA encoded by said gene, or biological material containing said gene or said miRNA in molecular breeding or breed improvement of cashmere goats; the nucleotide sequence of said chi-miR-1388-3p gene is shown in any of the following:

[0017] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0018] (2) The nucleotide sequence complementary to (1);

[0019] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0020] Preferably, the biological material is recombinant DNA, expression cassette, transposon, plasmid vector, phage vector, viral vector, engineered bacteria, or transgenic cell line.

[0021] Preferably, the application includes: promoting the proliferation of hair papilla cells in cashmere goat skin after inhibiting the chi-miR-1388-3p gene, or increasing the ratio of S-phase to G2 / M-phase cells in cashmere goat skin, or inhibiting apoptosis of hair papilla cells in cashmere goat skin.

[0022] Preferably, the application includes: overexpressing the chi-miR-1388-3p gene to inhibit the proliferation of hair papilla cells in cashmere goat skin, or to reduce the ratio of S-phase to G2 / M-phase cells in cashmere goat skin, or to promote apoptosis of hair papilla cells in cashmere goat skin.

[0023] Preferably, melatonin is used to suppress the chi-miR-1388-3p gene.

[0024] Preferably, all of the above applications are melatonin-mediated.

[0025] Fifthly, this invention provides the application of melatonin in inhibiting the chi-miR-1388-3p gene;

[0026] The nucleotide sequence of the chi-miR-1388-3p gene is shown in any of the following:

[0027] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0028] (2) The nucleotide sequence complementary to (1);

[0029] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0030] In a sixth aspect, the present invention provides a method for regulating the proliferation, cell cycle or apoptosis of hair papilla cells in cashmere goat skin, comprising: controlling the expression of the chi-miR-1388-3p gene in hair papilla cells in cashmere goat skin;

[0031] The nucleotide sequence of the chi-miR-1388-3p gene is shown in any of the following:

[0032] (1) The nucleotide sequence shown in SEQ ID NO.1;

[0033] (2) The nucleotide sequence complementary to (1);

[0034] (3) A nucleotide sequence with one or more nucleotides replaced, deleted and / or added to the nucleotide sequence shown in SEQ ID NO.1 and having the same function.

[0035] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0036] This invention identifies and analyzes miRNAs in the melatonin-implanted skin tissue of Inner Mongolian cashmere goats in the early growth stage using transcriptome sequencing technology. Through bioinformatics analysis, differentially expressed miRNAs that significantly affect cashmere growth and hair follicle development are screened out, and their functions are explored at the cellular level.

[0037] This invention discovers a microRNA (chi-miR-1388-3p) associated with the growth cycle of cashmere goat follicles, which can regulate the proliferation, cell cycle, or apoptosis of dermal papilla cells in cashmere goat skin, thereby regulating cashmere goat follicle growth and cashmere growth. Furthermore, this invention is the first to discover that melatonin-mediated interference with chi-miR-1388-3p promotes the proliferation of dermal papilla cells in cashmere goat skin, regulates the cell cycle, and inhibits apoptosis, providing valuable insights into the regulatory role of melatonin-mediated miRNAs in cashmere goat follicle growth. Attached Figure Description

[0038] Figure 1 This is a statistical analysis of small RNA length in the melatonin implantation group (VPT1)(A) and the control group (VPC1)(B).

[0039] Figure 2 This is an analysis of differentially expressed miRNAs, where A is a Venn diagram of differentially expressed miRNAs and B is a volcano diagram of differentially expressed miRNAs.

[0040] Figure 3 This is a GO enrichment analysis of differentially expressed miRNA target genes in the control group and the implantation group.

[0041] Figure 4 This is a KEGG enrichment analysis of differentially expressed miRNA target genes in the control group and the implantation group, focusing on signaling pathways.

[0042] Figure 5 A is a graph showing the transfection efficiency of dermal papilla cells after transfection with MT+chi-miR-1388-3p(hi), chi-miR-1388-3p(hi), melatonin (MT), and control (NC);

[0043] Figure 5 B represents the transcriptional levels of the chi-miR-1388-3p gene detected by qRT-PCR in different treatment groups.

[0044] Figure 6 The results show the effects of transfection with chi-miR-1388-3p(hi), MT+chi-miR-1388-3p(hi), melatonin (MT), and control (NC) on the proliferation of skin dermal papilla cells.

[0045] Figure 7 The results of flow cytometry analysis of dermal papilla cell cycle after transfection with chi-miR-1388-3p(hi), MT+chi-miR-1388-3p(hi), melatonin (MT), and control (NC) are as follows. Figure 7 Figure A shows the cell counting results at different cell cycles. Figure 7 B is a statistical graph of cell counts at different cell cycles.

[0046] Figure 8 The results show the effects of transfection with chi-miR-1388-3p(hi), MT+chi-miR-1388-3p(hi), melatonin (MT), and control (NC) on apoptosis of skin dermal papilla cells. Figure 8 A represents the results of flow cytometry analysis. Figure 8 B is a statistical chart of flow cytometry results. Detailed Implementation

[0047] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0048] Unless otherwise specified, all methods used in the examples were conventional or performed according to techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents and instruments used without specified manufacturers were all conventional products that could be purchased from legitimate channels.

[0049] Example 1: Transcriptome sequencing of skin tissue from early growth stage of Inner Mongolian cashmere goats induced by melatonin.

[0050] 1. We mixed the total RNA from goat skin tissues in each group (n=3) to reduce individual differences among goats, and constructed two miRNA pool libraries from the melatonin implantation group and the control group. The construction method for the melatonin implantation group and the control group of Inner Mongolian cashmere goats was as follows: Liu J, Mu Q, Liu Z, Wang Y, Liu J, Wu Z, Gong W, Lu Z, Zhao F, Zhang Y, Wang R, Su R, Li J, Xiao H, Zhao Y. Melatonin Regulates the Periodic Growth of Cashmereby Upregulating the Expression of Wnt10b and β-catenin in Inner Mongolia Cashmere Goats. Front Genet. 2021 Jul 9; 12:665834. Each pool contained 2 μg of RNA to generate two RNA pool samples, and sequencing libraries for the cashmere goat control group and the implantation group were constructed. miRNA sequencing library construction: Following the manufacturer's instructions, a miRNA sequencing library was constructed using the TruSeq Small RNA Sample Preparation Kit (Illumina, San Diego, CA, USA). Starting with total RNA, sRNA transcripts were ligated to the 3′ and 5′ ends with appropriate aptamers. cDNA was synthesized using reverse transcription, followed by stepwise PCR amplification of the product. Target DNA fragments were then separated using gel electrophoresis (PAGE), and the cDNA library was obtained after gel extraction and recovery. Single-end sequencing of the library (50 bp) was performed on an Illumina HiSeq 2500 platform.

[0051] Raw reads were obtained from sequencing, and then the following series of data quality assessments were performed on the obtained raw reads: (1) low-quality reads (reads with a quality value of sQ≤20 accounting for more than 30% of the total read length); (2) reads with a proportion of bases whose information could not be determined >10% were removed; (3) reads with 5' adapter contamination were removed; (4) reads without 3' adapter sequences and insert fragments were removed; (5) 3' adapter sequences were removed; (6) reads containing polyA / T / G / C were removed to purify the reads.

[0052] 2. Bowtie was used to align the screened small RNAs with goat reference sequences, and reads matching the goat reference sequences were compared with sequences within a specified range in miRbase. This yielded detailed information on the small RNAs in each sample, including matching known and potential miRNAs and their secondary structures, as well as the sequence, length, frequency of occurrence, and base distribution of each miRNA in each sample. Further statistical analysis of the length distribution of the high-quality data revealed that the lengths of the miRNAs in both groups ranged from 18 to 35 nt, with most miRNA sequences concentrated between 20 and 24 nt. The lengths of the mature miRNA sequences in both the implantation and control groups were mostly 22 nt. See below for details. Figure 1 distributed.

[0053] Clean reads were aligned to the Rfam database, and rRNA, tRNA, snRNA, and snoRNA sequences were removed based on annotations. Small RNAs were aligned to the exons and introns of goat genome mRNA sequences, and sRNAs derived from mRNA degradation fragments were removed.

[0054] We utilized the hairpin structure of miRNA precursors, combined with miREvo and miRdeep2 software, to analyze and predict novel miRNAs. To summarize the unique annotation of each small RNA type, small RNAs were compared sequentially according to the following priority rule: known miRNA > rRNA > tRNA > snRNA > snoRNA > repeat > gene > novel miRNA.

[0055] 3. Known mRNAs and miRNAs were normalized using TPM (Transcripts Per Million Reads). Differential expression analysis was performed on the two groups using DEGSeq, and differentially expressed miRNAs were screened based on |log2(Fold Change)| ≥ 1 and P < 0.05. (See [link to documentation]). Figure 2 As shown.

[0056] 4. GOseq was used to perform functional enrichment analysis on the target genes of differentially expressed miRNAs in the skin tissues of the Inner Mongolia cashmere goat control and implantation groups. KOBAS software was used to perform KEGG signaling pathway enrichment analysis on differentially expressed genes based on the KEGG (Kyoto Encyclopedia of Genes and Genomes, KEGG) database. (See attached image.) Figure 3 and Figure 4 As shown.

[0057] GO functional analysis revealed that target genes are mainly involved in transmembrane transport, protein modification, and cellular protein modification processes in biological processes. Within cellular components, target genes are primarily involved in the cytoskeleton, cytoskeleton, and extracellular domains. In terms of molecular function, target genes are mainly involved in binding, protein binding, and molecular functional regulation. KEGG enrichment analysis of differentially expressed miRNA target genes showed significant enrichment in the MAPK signaling pathway, Notch signaling pathway, and Ras signaling pathway, which play important roles in hair follicle growth and cell cycle. Furthermore, a series of pathways related to cell proliferation, differentiation, and migration, including endocytosis, GnRH signaling pathway, and carbon metabolism, were also identified.

[0058] 5. Based on comprehensive analysis of gene expression levels, GO and KEGG analysis results, differentially expressed chi-miR-1388-3p was screened as a candidate gene, the sequence of which is shown in SEQ ID NO.1: AUCUCAGGUUCGUCAGCCCGCA.

[0059] Example 2: Effects of melatonin-mediated chi-miR-1388-3p on dermal papilla cell proliferation and apoptosis

[0060] 1. Isolation and culture of hair papilla cells (DPCs) from cashmere goats

[0061] In this laboratory, dermal papilla cells from the skin of Inner Mongolian cashmere goats were isolated. First, the scapular skin tissue was disinfected with 75% alcohol and then washed with PBS buffer. The tissue sections were then cut into 1mm pieces using sterile surgical scissors. 2 Small fragments were collected and placed in a culture dish containing 0.25% neutral protease, and cultured at 37°C for approximately 2 hours. The cells were dissected under a microscope, and the dermis and subcutaneous tissue were carefully removed using micro-forceps. The dermal papilla was extracted from the base of the hair follicle. The separated dermal papilla cells adhered to the culture dish and were cultured in DMEM medium containing 10% FBS and 1% penicillin / streptomycin at 37°C, with the medium changed every 2 to 3 days. The cells were passaged once when they began to emerge from the edge of the tissue fragment and the cell density reached 80-90%.

[0062] 2. Construction and packaging of lentiviral vectors

[0063] (1) Construction of lentiviral vectors: Lentiviral vectors for the negative control (NC) group and the HBLV-chi-miR-1388-3p-Null-ZsGreen-PURO (chi-miR-1388-3p[hi]) group were purchased from Shanghai Hanheng Biotechnology Co., Ltd.

[0064] (2) Packaging of lentiviruses: The constructed lentivirus vector and helper plasmids were extracted in large quantities, and viruses with an A260 / A280 value of about 1.75 and a concentration of >1 μg / μL were packaged and transfected.

[0065] (3) Filtration and preservation of lentivirus: Change the medium 6 h after transfection, and collect the virus supernatant at 48 h and 72 h respectively (collect the supernatant 48 h after transfection and add fresh medium). After collection, filter into 40 mL centrifuge tubes through a 0.45 μm filter, centrifuge for 120 min (4℃, 7200 rpm), resuspend the virus pellet in 500 μL of fresh medium, and store at -80℃.

[0066] 3. Lentiviral transfection of target cells

[0067] (1) Cell preparation: Healthy dermal papilla cells were seeded into 24-well cells at a seeding density of 2 × 10⁶ cells / well. 4 Cells were incubated at 37°C and 5% CO2 overnight. The seeding number varied slightly depending on the cell growth rate, but generally, the cells were transfected after reaching a confluence of 30-50%. Before transfection, each experimental group was incubated with Polybrene dilution buffer to a final concentration of 5 μg / mL.

[0068] (2) Viral infection: After incubation for 4 hours, the original culture medium was discarded and the 1 / 2 small volume infection method was used, that is, 1 / 2 volume of fresh culture medium was added during viral infection. For the experimental group containing melatonin treatment, 10 μL of melatonin solution with a concentration of 300 pg / mL was added to the cell culture medium. After 4 hours of lentivirus infection, the remaining 1 / 2 culture medium was added. The multiplicity of infection for each experimental group was 5.

[0069] (3) Change the medium: 24 hours after infection, remove the culture medium containing the virus, replace it with fresh complete culture medium, and continue to culture in a 37°C, 5% CO2 incubator.

[0070] (4) Observe fluorescence: 48h after transfection, the expression efficiency of GFP can be initially observed by fluorescence microscopy for viruses carrying the GFP reporter gene. Generally, the expression peak can be reached 72h after infection and transfection. The transfection efficiency is observed by fluorescence microscopy. Puromycin is added to each experimental group for resistance screening to screen stable transduced cell lines. The final concentration is 5μg / mL.

[0071] Fluorescence microscopy showed successful antibiotic resistance screening; the cells exhibited strong, uniformly distributed green fluorescence, indicating successful construction of a stable cell line. Figure 5 A).

[0072] qRT-PCR was used to detect the transcriptional level of the chi-miR-1388-3p gene in different treatment groups. Different volumes of reverse transcription reagent were added according to the RNA concentration to be measured (see Table 1). The reverse transcription program was 42℃ for 60 min and 70℃ for 10 min. The qPCR reaction system is shown in Table 2, and the qPCR reaction program is shown in Table 3. 2 -ΔΔCt The relative expression levels of miRNAs were calculated and assessed. Data are expressed as mean ± standard deviation (Mean ± SD), and each experiment was performed in triplicate.

[0073] Table 1 miRNA reverse transcription reaction system

[0074]

[0075] Table 2 miRNA qPCR reaction system

[0076]

[0077] Table 3 miRNA qPCR reaction procedure

[0078]

[0079] qRT-PCR results showed that, compared with the melatonin-treated group (MT), the expression level of chi-miR-1388-3p in DPCs was significantly increased after transfection with [MT+chi-miR-1388-3p(hi)] (P < 0.01). Compared with the negative control group (NC), the expression level of chi-miR-1388-3p in the melatonin group (MT) was significantly decreased (P < 0.01), indicating that melatonin negatively regulates chi-miR-1388-3p in DPCs. This result is consistent with previous sequencing results. Figure 5 B.

[0080] 4. CCK-8 assay for Detection of Hair Papillary Cell Proliferation

[0081] Cell proliferation was assessed using the CCK-8 assay. Five × 10⁵ cells were seeded per well of a 96-well plate containing dermal papilla cells. 5Cells were incubated at 10 μL of CCK-8 reagent in each well at 0, 24, 48, and 72 hours after transfection with lentivirus, followed by incubation for two hours. The optical density (OD) of each well was measured using a microplate reader at 0, 24, 48, and 72 hours. The measured OD values ​​were calculated and plotted using GraphPad 8.0. The results showed that compared with the negative control group (NC) and the melatonin treatment group (MT), the melatonin and chi-miR-1388-3p overexpression co-treatment group significantly inhibited cell proliferation at 72 hours (P < 0.01). Compared with the chi-miR-1388-3p overexpression group alone, the melatonin + chi-miR-1388-3p overexpression co-treatment group significantly inhibited cell proliferation from 48 to 72 hours (P < 0.01). These data indicate that melatonin-mediated downregulation of chi-miR-1388-3p promotes dermal papilla cell proliferation. (See [link to article]). Figure 6 .

[0082] 5. Flow cytometry detection of dermal papilla cell cycle

[0083] Hair papilla cells were arranged at 5 × 10⁶ cells per pore. 5 Cells were seeded at a density of [number] cells / well in 6-well plates. Forty-eight hours after transfection, the collected culture was digested with trypsin for 5 minutes, followed by centrifugation at 1000×g for 5 minutes. Cells were then resuspended in 1 mL of pre-chilled PBS, centrifuged again at 1000×g for 5 minutes, the supernatant was removed, and 5 μL of pre-chilled 70% ethanol was added for fixation for 12 h. The cell fixative was washed with PBS, and a cell suspension was prepared by adding 400 μL of propidium iodide (PI) staining buffer. The cells were then incubated at 37°C in the dark for 30 minutes. Flow cytometry was used to detect red fluorescence at a laser wavelength of 488 nm to analyze the cell suspension. The results showed that melatonin-mediated downregulation of chi-miR-1388-3p increased the proportion of dermal papilla cells in S and G2 / M phases and decreased the proportion of cells in G0 / G1 phase. [See attached image]. Figure 7 .

[0084] 6. Flow cytometry detection of dermal papilla cell apoptosis

[0085] Prepare a single-cell suspension and centrifuge at 1000g for 3-5 min at 4℃. Collect cells, wash twice with pre-cooled PBS, centrifuge again, discard the supernatant, and resuspend the cells in 500 μL of diluted 1×Annexin V Binding Buffer. Add 5 μL of Annexin V-APC Reagent and 5 μL of PI reagent (50 μg / mL) to the cell suspension. Gently rotate the cells and incubate in the dark for 15 min. Immediately analyze the results. The results show that melatonin mediates downregulation of chi-miR-1388-3p, inhibiting apoptosis of dermal papilla cells. (See [link to study]). Figure 8 .

[0086] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of chi-miR-1388-3p in inhibiting the proliferation of hair papilla cells in cashmere goat skin, increasing the proportion of G0 / G1 phase cells in cashmere goat skin, decreasing the proportion of S phase and G2 / M phase cells in cashmere goat skin, or promoting apoptosis of hair papilla cells in cashmere goat skin; the nucleotide sequence of chi-miR-1388-3p is shown in SEQ ID NO.

1.

2. The application according to claim 1, characterized in that, The applications include: inhibiting the proliferation of hair papilla cells in cashmere goat skin after overexpression of chi-miR-1388-3p, or reducing the ratio of S-phase to G2 / M-phase cells in cashmere goat skin, or promoting apoptosis of hair papilla cells in cashmere goat skin.

3. A method for regulating the proliferation, cell cycle, or apoptosis of hair papilla cells in cashmere goat skin, characterized in that, include: Overexpression of chi-miR-1388-3p can inhibit the proliferation of dermal papilla cells in cashmere goat skin, or increase the proportion of dermal papilla cells in the G0 / G1 phase, or decrease the proportion of dermal papilla cells in the S phase and G2 / M phase, or promote apoptosis of dermal papilla cells in cashmere goat skin; the nucleotide sequence of chi-miR-1388-3p is shown in SEQ ID NO.1.

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