环状RNA ciRS-7靶向miR-219a-5p影响微小隐孢子虫体外增殖的机制

doi:10.11843/j.issn.0366-6964.2022.11.024

Abstract

Abstract: The present study was carried out to investigate the impact of circRNA ciRS-7 on the propagation of Cryptosporidium parvum in HCT-8 cells via targeting miR-219a-5p. Using C. parvum-infecting HCT-8 cells as the model, we detected the expression of the autophagy associated-proteins of LC3B-II and p62 in HCT-8 cells by using Western bolt assays. The targeting relationship between ciRS-7 and miR-219a-5p were detected and verified by using qRT-PCR and dual luciferase reporter assays, and the infection burden of C. parvum (the mRNA level of C. parvum hsp70 gene) in HCT-8 cells was determined by using qRT-PCR. C. parvum infection induced significantly up-regulation of the protein levels of LC3B-II and p62 in HCT-8 cells at 8 h, 12 h, 24 h, 36 h and 48 h post-infection. Overexpression of ciRS-7 markedly inhibited the protein level of LC3B-II but promoted the protein level of p62 in C. parvum-infecting cells, whereas interfering the expression of ciRS-7 obtained the opposite results. C. parvum infection induced significant down-regulation of miR-219a-5p in HCT-8 cells, and ciRS-7 could target to regulate the expression of miR-219a-5p. miR-219a-5p mimics significantly increased the protein level of LC3B-II but decreased the protein level of p62 in C. parvum-infecting cells, while miR-219a-5p inhibitor had the opposite effects, and the inhibition effect on the protein level of LC3B-II and the promoted effect on the protein level of p62 by overexpression of ciRS-7 in C. parvum-infecting cells were reversed by miR-219a-5p mimics. miR-219a-5p mimics significantly inhibited the mRNA level of C. parvum hsp70 in infected cells, while miR-219a-5p inhibitor had the opposite effect. The upregulated effect of overexpression of ciRS-7 on the mRNA level of C. parvum hsp70 gene in infected cells was attenuated by miR-219a-5p mimics. The results revealed that ciRS-7 promoted the propagation of C. parvum in HCT-8 cells by targeting miR-219a-5p to inhibit C. parvum-induced cell autophagy.

Key words: Cryptosporidium parvum, circRNA, ciRS-7/miR-219a-5p axis, autophagy, propagation

CLC Number:

S852.723

YIN Yanling, HUANG Shuang, YAO Qian, WU Jiangping, GUO Haochen, YANG Xin, SONG Junke, ZHAO Guanghui. The Mechanisms of Circular RNA ciRS-7 Affecting the Propagation of Cryptosporidium parvum in HCT-8 Cells via Targeting miR-219a-5p[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3989-3999.

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The Mechanisms of Circular RNA ciRS-7 Affecting the Propagation of Cryptosporidium parvum in HCT-8 Cells via Targeting miR-219a-5p

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