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Effect of miR-27b-3p and Nrf2 in human retinal pigment epithelial cell induced by high-glucose

International Journal of Ophthalmology(English edition) 2023年第10期16卷 1582-1588页

作者： Qiao-Ling Lai Ting Xie Wei-Dong Zheng Yan Huang Department of Ophthalmology and Optometry Fujian Medical UniversityFuzhou 350004Fujian ProvinceChina The First Affiliated Hospital of Fujian Medical University Fuzhou 350004Fujian ProvinceChina

AIM:To determine whether the microrna-27b-3p(miR-27b-3p)/NF-E2-related factor 2(Nrf2)pathway plays a role in human retinal pigment epithelial(hRpE)cell response to high glucose,how miR-27b-3p and Nrf2 expression are regulated,and whether this pathway could be specifically ***:hRpE cells were cultured in normal glucose or high glucose for 1,3,or 6d before measuring cellular proliferation rates using cell counting kit-8 and reactive oxygen species(ROS)levels using a dihydroethidium ***-27b-3p,Nrf2,NAD(p)H quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)mRNA and protein levels were analyzed using reverse transcription quantitative polymerase chain reaction(RT-qpCR)and immunocytofluorescence(ICF),*** blot analyses were performed to determine nuclear and total Nrf2 protein ***2,NQO1,and HO-1 expression levels by RT-qpCR,ICF,or Western blot were further tested after miR-27b-3p overexpression or inhibitor lentiviral ***,the expression level of those target genes was analyzed after treating hRpE cells with ***:persistent exposure to high glucose gradually suppressed hRpE Nrf2,NQO1,and HO-1 mRNA and protein levels and increased miR-27b-3p mRNA *** glucose also promoted ROS release and inhibited cellular ***2,NQO1,and HO-1 mRNA levels decreased after miR-27b-3p overexpression and,conversely,both mRNA and protein levels increased after expressing a miR-27b-3p *** treating hRpE cells exposed to high glucose with pyridoxamine,ROS levels tended to decreased,proliferation rate increased,Nrf2,NQO1,and HO-1 mRNA and protein levels were upregulated,and miR-27b-3p mRNA levels were ***:Nrf2 is a downstream target of ***,the miR-27b-3p inhibitor pyridoxamine can alleviate high glucose injury by regulating the miR-27b-3p/Nrf2 axis.

关键词： human retinal pigment epithelial cell high glucose pyridoxamine microrna-27b-3p NF-E2-related factor 2 NAD(p)H quinone oxidoreductase 1 heme oxygenase-1

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LncRNA AFAp1-AS1/miR-27b-3p/VEGF-C axis modulates stemness characteristics in cervical cancer cells

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Chinese Medical Journal 2021年第17期134卷 2091-2101页

作者： Meng Xia Li-Jun Duan bi-Nan Lu Yu-Zhou pang Zong-Ran pang School of pharmacy Minzu University of ChinaBeijing 100081China Department of Orthopedics Bayannaoer City HospitalBayannaoerInner Mongolia 015000China Guangxi Zhuang Yao Medicine Center of Engineering and Technology Guangxi University of Chinese MedicineNanningGuangxi 530200China Key Laboratory of Ethnomedicine of Ministry of Education Minzu University of ChinaBeijing 100081China

background:Long non-coding RNA(lncRNA)actin filament-associated protein 1 antisense RNA 1(AFAp1-AS1)functions as a competing endogenous RNA to regulate target genes expression by sponging micrornas(miRs)to play cancer-promoting roles in cancer stem ***,the regulatory mechanism of AFAp1-AS1 in cervical cancer(CC)stem cells is *** present study aimed to provide a new therapeutic target for the clinical treatment of ***:Hyaluronic acid receptor cluster of differentiation 44 variant exon 6(CD44v6)(+)CC cells were isolated by flow cytometry(FCM).Small interfering RNAs of AFAp1-AS1(siAFAp1-AS1)were transfected into the(CD44v6)(+)*** levels of AFAp1-AS1 were measured by quantitative real-time pCR(qRT-pCR).Sphere formation assay,cell cycle analysis,and Western blotting were used to detect the effect of *** pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAp1-AS1 or vascular endothelial growth factor(VEGF)-***:CD44v6(+)CCcells had remarkable stemness and a high level ***,AFAp1-AS1knockdownwithsiAFAp1-AS1suppressed the cell cycle transitionofG(1)/S phase and inhibited self-renewal ofCD44v6(+)CCcells,the levels of the stemnessmarkers octamer-binding transcription factor 4(OCT4),osteopontin(OpN),and cluster of differentiation 133(CD133),and the epithelialmesenchymal transition(EMT)-related proteins Twist1,matrix metalloprotease(MMp)-9,and *** the mechanism study,miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAp1-AS1 in the CD44v6(+)CC ***:LncRNA AFAp1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.

关键词： Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 Cell stemness Cervical cancer Long non-coding RNA actin filament-associated protein 1 antisense RNA 1 microrna-27b-3p

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