LncRNA AFAP1-AS1/miR-27b-3p/VEGF-C axis modulates stemness characteristics in cervical cancer cells

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 cells.However,the regulatory mechanism of AFAP1-AS1 in cervical cancer(CC)stem cells is unknown.The present study aimed to provide a new therapeutic target for the clinical treatment of CC.Methods: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)(+)cells.The 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 siAFAP1-AS1.RNA 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)-C.Results:CD44v6(+)CCcells had remarkable stemness and a high level ofAFAP1-AS1.However,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 VEGF-C.In the mechanism study,miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+)CC cells.Conclusions:LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.

Hypoglycemic effects of Tibetan medicine Huidouba in STZ-induced diabetic mice and db/db mice

Objective:Huidouba(HDB) is a Chinese folk medicine used to treat diabetes in Sichuan Province,China.Therefore,we investigated the anti-diabetic effects of HDB and its underlying mechanisms.We hypothesized that HDB treatment could enhance glucose tolerance and insulin sensitivity,and thus prevent a hyperglycemia state.Methods:To test the hypothesis,streptozotocin(STZ)-induced diabetic mice and db/db mice,widely used models of hyperglycemia and insulin-resistant diabetes,were either treated with HDB,metformin,or acarbose.Blood glucose,oral glucose tolerance test,insulin tolerance test,pancreatic histopathology and serum biochemistry were detected to assess the hypoglycemic effect of HDB.Results:HDB treatments were found to show the effect in reducing glucose levels.HDB also resulted in a significant reduction in body weight and food intake in the STZ-induced diabetic mouse model.Furthermore,it significantly improved glucose and insulin tolerance in the two diabetic mouse models.Importantly,insulin,glucagon,pancreatic polypeptide,and somatostatin immunohistochemistry revealed that HDB treatment improved the function and the location of the cells in the islets compared with the other two treatments.HDB treatment resulted in significant restoration of islet function.Our results illustrated the underlying mechanism of HDB in the progression of diabetes,and HDB can be an effective agent for the treatment of diabetes.Conclusion:The results of this study suggested that HDB can reduce blood glucose levels in STZ-induced hyperglycemic mice and db/db mice.