The effect of lncRNA FEZF1-AS1 targeting regulation of miR-200c-3p on biological behaviors of human lung fibroblasts

doi:10.11958/20230719

Abstract

Abstract:

Objective To investigate the effect of FEZ family zinc finger 1-antisense RNA 1 (LncRNA FEZF1-AS1) targeting regulation of miR-200c-3p expression on biological behaviors of human lung fibroblasts (HLF). Methods Transforming growth factor β1 (TGF-β1) was used to induce the transformation of HLF into myofibroblasts, which were divided into the Blank group and the model group (HLF+TGF-β1 group). According to different transfection plasmid, cells were divided into the Blank group, the TGF-β1+Si LncRNA FEZF1-AS1 NC group and the TGF-β1+Si LncRNA FEZF1-AS1 group. The protein expressions of α-SMA, Collagen Ⅰ and Vimentin were detected by Western blot assay. The expressions of LncRNA FEZF1-AS1 and miR-200c-3p were detected by quantitative real-time PCR (qRT-PCR). Cell proliferation ability was detected by CCK-8 method, migration ability was detected by cell scratch experiment and invasion ability was detected by Transwell assay. The targeting relationship between FEZF1-AS1 and miR-200c-3p was detected by dual-luciferase reporter assay. Results Compared with the Blank group, protein expressions of α-SMA, Collagen Ⅰ, Vimentin and the expression of LncRNA FEZF1-AS1 were increased in the HLF+TGF-β1 group (P<0.05), and the expression of miR-200c-3p was decreased (P<0.05). Compared with the TGF-β1+Si LncRNA FEZF1-AS1 NC group, cell proliferation, migration, invasion ability, LncRNA FEZF1-AS1 expression, protein expressions of α-SMA, Collagen Ⅰ and Vimentin were decreased in the TGF-β1+Si LncRNA FEZF1-AS1 group (P<0.05), and the expression of miR-200c-3p was increased (P<0.05). There were binding sites between miR-200c-3p and FEZF1-AS1 gene sequence. Conclusion LncRNA FEZF1-AS1 promotes the formation and progression of idiopathic pulmonary interstitial fibrosis by inhibiting miR-200c-3p.

Key words: idiopathic pulmonary interstitial fibrosis, lung fibroblasts, myofibroblasts, FEZ family zinc finger 1-antisense RNA1, microRNA-200c-3p

CLC Number:

R563.9

Figures/Tables 11

References 17

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基因名称	引物序列（5'→3'）	产物大小/bp
LncRNA FEZF1-AS1	上游：TTAGGAGGCTTGTTCTGTGT	239
LncRNA FEZF1-AS1	下游：GCGCAGGTACTTAAGAAAGA	239
miR-200c-3p	上游：UAAUACUGCCGGGUAAUGAUGGA	70
	下游：UCCAUCAUUACCCGGCAGUAUUA	70
GAPDH	上游：GCCTTCCGTGTCCCCACTGC	216
	下游：GGCTGGTGGTCCAGGGGTCT	216
U6	上游：CTCGCTTCGGCAGCACA	65
	下游：AACGCTTCACGAATTTGCGT	65

基因名称	引物序列（5'→3'）	产物大小/bp
LncRNA FEZF1-AS1	上游：TTAGGAGGCTTGTTCTGTGT	239
LncRNA FEZF1-AS1	下游：GCGCAGGTACTTAAGAAAGA	239
miR-200c-3p	上游：UAAUACUGCCGGGUAAUGAUGGA	70
	下游：UCCAUCAUUACCCGGCAGUAUUA	70
GAPDH	上游：GCCTTCCGTGTCCCCACTGC	216
	下游：GGCTGGTGGTCCAGGGGTCT	216
U6	上游：CTCGCTTCGGCAGCACA	65
	下游：AACGCTTCACGAATTTGCGT	65

组别	α-SMA	Collagen Ⅰ	Vimentin
Blank组	0.68±0.02	0.37±0.05	0.66±0.06
HLF+TGF-β1组	0.90±0.02	0.68±0.05	0.98±0.04
t	7.621^＊	7.431^＊	4.583^＊

组别	α-SMA	Collagen Ⅰ	Vimentin
Blank组	0.68±0.02	0.37±0.05	0.66±0.06
HLF+TGF-β1组	0.90±0.02	0.68±0.05	0.98±0.04
t	7.621^＊	7.431^＊	4.583^＊

组别	0 h	24 h	48 h	F
Blank组	0.49±0.00	0.65±0.01^A	1.08±0.02^AB	640.000^＊
TGF-β1+Si LncRNA FEZF1-AS1 NC组	0.48±0.01	1.08±0.03^aA	1.87±0.03^aAB	641.500^＊
TGF-β1+Si LncRNA FEZF1-AS1组	0.47±0.02	0.84±0.01^abA	1.53±0.07^abAB	185.300^＊
F	0.719	108.400^＊	79.310^＊