mdv1-miR-M4-5p对MDCC-MSB1细胞增殖和凋亡的影响

doi:10.11843/j.issn.0366-6964.2024.08.039

Abstract

Abstract:

The purpose of this experiment was to study the effects of mdv1-miR-M4-5p analog encoded by Marek's disease virus (MDV) on proliferation and apoptosis of MDCC-MSB1 cells. In this study, mdv1-miR-M4-5p mimics, inhibitors and their negative controls were transfected into MSB1 cells for 48 h. Real-time quantitative fluorescent PCR (qRT-PCR) was used to detect the transcription of mdv1-miR-M4-5p, TGF-β1, Smad2, caspase-9, caspase-3, cyt-c, cyclinD1, Bcl-2 et al other molecules related to proliferation and apoptosis in MDCC-MSB1 cells. Cell proliferation was detected by CCK-8 and cell cycle and apoptosis were detected by flow cytometry. The results showed that compared with the control group, the transcription levels of mdv1-miR-M4-5p, cyclinD1 and Bcl-2 in MDCC-MSB1 cells transfected with mdv1-miR-M4-5p mimics was significantly up-regulated. The transcription of TGF-β1, Smad2, cyt-c, caspase-9 and caspase-3 was down-regulated, the proliferation of MDCC-MSB1 cells was promoted, the the number of G1 phase cells were decreased, the S and G2 cells were increased, and the apoptosis rate was decreased. Compared with the control group, the results of the proliferation and apoptosis of MDCC-MSB1 cells and the expression of related molecules transfected with mdv1-miR-M4-5p inhibitor were opposite to those transfected with mdv1-miR-M4-5p mimics. The results showed that Mdv1-miR-M4-5p could promote the proliferation and inhibit the apoptosis of MDCC-MSB1 cells. This may be conducted by inhibiting the TGF-β1/Smad2 signaling pathway.

Key words: mdv1-miR-M4-5p, MDV, proliferation, apoptosis, TGF-β1/Smad2

CLC Number:

Zuhua YU, Mengru GAO, Lei HE, Ying WEI, Jian CHEN, Songbiao CHEN, Ke DING. Effects of mdv1-miR-M4-5p Encoded by MDV on Proliferation and Apoptosis of MDCC-MSB1 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3678-3687.

Figures/Tables 5

Table 1

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名称 Name	引物序列(5′→3′) Sequence	用途 Application
U6 RT prime	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCGATACA	反转录
U6 forward	CGCTTCGGCAGCACATATAC	Real-time PCR
miR-M4-5p RT prime	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGAAGGGTT	反转录
miR-M4-5p forward	TGCGCTTAATGCTGTATCGGAAC	Real-time PCR
miR-M4-5p/U6 reverse	CCAGTGCAGGGTCCGAGGTATT	Real-time PCR
GAPDH forward	CCAGAACATCATCCCAGCGT	Real-time PCR
GAPDH reverse	CAGGTCAGGTCAACAACAGA	Real-time PCR
TGF-β1 forward	GTATTGGGCCAAAGAGCTGC	Real-time PCR
TGF-β1 reverse	ATTGCCGTAACCCTGGTACA	Real-time PCR
Smad2 forward	GTGGCAGATGAGGGTGATTT	Real-time PCR
Smad2 reverse	TGCAGCCACAGGACGAAAC	Real-time PCR
Caspase-3 forward	TGTGGACTCTGGAATTCTGC	Real-time PCR
Caspase-3 reverse	TGCCTGAACGAGATGACAGT	Real-time PCR
Caspase-9 forward	CCGAAGGGAGCAAGCACG	Real-time PCR
Caspase-9 reverse	AGGTTGGACTGGGATGGAC	Real-time PCR
Cyt-c forward	AGGCAAGCACAAGACTGGA	Real-time PCR
Cyt-c reverse	CTGACTATCACCAAGAACCACC	Real-time PCR
Bcl-2 forward	TACCAGAGGGACTTCGC	Real-time PCR
Bcl-2 reverse	TTGACCCCATCACGGA	Real-time PCR
cyclinD1 forward	AGGTTGGACTGGGATGGAC	Real-time PCR
cyclinD1 reverse	TGCAGCCACAGGACGAAACAG	Real-time PCR

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Effects of mdv1-miR-M4-5p Encoded by MDV on Proliferation and Apoptosis of MDCC-MSB1 Cells

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