MiR-361-5p/abca1 and MiR-196-5p/arhgef12 Axis Involved in γ-Sitosterol Inducing Dual Anti-Proliferative Effects on Bronchial Epithelial Cells of Chronic Obstructive Pulmonary Disease

Abstract

Chronic obstructive pulmonary disease (COPD), a progressive and irreversible respiratory disease, becomes the third leading cause of death and results in enormous economic burden on healthcare costs and productivity loss worldwide by 2020. Thus, it is urgent to develop effective anti-COPD drugs. Materials and Methods: In the present study, two published GEO profiles were used to reanalyze and ascertain the relationships between circulating miRNAs and bronchial epithelial cells (BECs) mRNAs in COPD. The microRNA levels of miR-361-5p and miR-196-5p in plasma of COPD patients and healthy volunteers were detected by qRT-PCR. Next, the effects of γ-sitosterol (GS) on the expression of miR-361-5p and miR-196-5p and cell proliferation were investigated in BEC and H292 cell lines. Finally, whether specific miRNA-mRNA pathways involved in the effect of GS on BECs was assayed using Western Blot, real-time PCR and immunofluorescence. Results: miR-196-5p and miR-361-5p were, respectively, up-and down-regulated in COPD patients compared with healthy controls. Luciferase assays demonstrated that miR-361-5p and miR-196-5p were, respectively, targeting abca1 and arhgef12 3ʹUTR in BEAS-2B cells. GS significantly suppressed miR-196-5p and promoted miR-361-5p levels in BEAS-2B cells and inhibited BECs proliferation in vitro. GS promoted miR-361-5p expression, which inhibited BCAT1 mRNA and protein levels and weaken mTOR-pS6K pathway, resulted in anti-proliferation in BEAS-2B cells. In addition, RhoA was activated by ARHGEF12 due to the inhibitory effect of miR-196-5p on arhgef12-3ʹUTR which was partially abolished by GS suppressing miR-196-5p expression. Activated RhoA further activated ROCK1-PTEN pathway and finally inhibited mTOR pathway, resulting in induced BECs proliferation. The antiproliferation effect of GS was not observed in H292 cells. Conclusion:These findings indicate that miR-361-5p/abca1 and miR-196-5p/arhgef12 axis mediated GS inducing dual anti-proliferation effects on BECs.

Seven ferroptosis-specific expressed genes are considered as potential biomarkers for the diagnosis and treatment of cigarette smoke-induced chronic obstructive pulmonary disease

“…We further searched the PubMed database (https:// pubmed.ncbi.nlm.nih.gov/) for literature related to miRNAs and COPD airway epithelium to screen for those overlapping with the above 281 miRNAs, excluding nonhuman specimen studies and studies without validation. This identified a total of 19 miRNAs: hsa-miR-130a-3p (24), hsa-miR-21 (25), hsa-miR-223-5p (26), hsa-miR-494-3p ( 27), miR-146a-5p (28), miR-221-3p (29), hsa-miR-92a-3p (29), miR-483-5p (30), hsa-miR-218-5p (31), miR-24-3p (32), miR-218 (33), miR-195 (34), hsa-miR-181c-5p (35), miR-16 (36), miR-222 (36), hsa-miR-29b-3p (37), hsa-miR-200c- 3p (38), hsa-miR-361-5p (39), and miR-146a (40). GO and KEGG enrichment analyses were performed on 7 differentially expressed ferroptosis hub genes.…”

Phytosterol and phytostanol-mediated epigenetic changes in cancer and other non-communicable diseases: a systematic review

“…2. The PSS reported were sitosterol (SITO) in five studies (37)(38)(39)(40)(41)(42) , Antcin-A (ATA) (43,44) and stigmasterol (STIG) (45,46) in two studies each and ergosterol (ERGO) (47) , and campesterol (CAMP) (45) in one each. Four studies completed experiments in breast cancer cell lines (38,39,43,48) , two in models of coronary obstructive pulmonary disease (42,47) , one in prostate cancer cell lines (40) , one in neural stem cells (37) , one in human aortic endothelial cells (41) , one in BALB/c-nude mice (38) , one was in guinea pigs (46) and one in silico (45) .…”

“…Similarly in HAEC, signalling pathway analysis indicated that SITO treatment enriched for AKT signalling, forkhead box O transcription factors and cell cycle signalling ( 37 ) . Contrary to this, the bronchial epithelial airway cell line model of chronic obstructive pulmonary disease (BEAS-2B) responded to very low concentrations of SITO (60 nM for 24 h) by induction of miR-196-5p and suppression miR-361-5p, leading to inhibition of mTOR signalling and reduced proliferation ( 42 ) .…”

“…In cancer, PSS induce miRNA that lower expression of factors such as ZEB1 that promote the epithelial to mesenchymal transition, a critical step in cancer metastasis ( 43 ) and suppress miRNA allowing increase in expression of factors that maintain an epithelial phenotype, such as CDH1 ( 43 ) . PSS also inhibit components of inflammatory pathways, including NF κ B and mTOR signalling, in models of coronary obstructive pulmonary disease ( 42 ) indicating a potential dietary route to ameliorating these conditions. Investigations into the epigenetic effects of PSS on many cell types, including those of the immune system, are lacking from the literature thus far.…”

“…The link between miRNA and PSS exposure was investigated in six records (37,38,(41)(42)(43)48) , of which two were screening studies (37,41) . The screening studies evaluated SITO in the context of rat neuronal stem cells (37) , and human aortic endothelial cells (41) .…”