Limits...
MicroRNA-153 is a prognostic marker and inhibits cell migration and invasion by targeting SNAI1 in human pancreatic ductal adenocarcinoma.

Bai Z, Sun J, Wang X, Wang H, Pei H, Zhang Z - Oncol. Rep. (2015)

Bottom Line: The mean expression of miR-153 in PDAC tissues was significantly reduced as compared to that in the normal pancreatic tissues.Notably, SNAI1 was identified as a direct target of miR-153 in PDAC.In conclusion, the results showed miR-153 is an independent prognostic marker for predicting survival in PDAC patients and inhibits cell migration and invasion by targeting SNAI1.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.

ABSTRACT
Human pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer type with early metastasis, which leads to poor prognosis for patients. Mounting evidence suggests that microRNAs (miRNAs) act as critical factors for tumor recurrence and metastasis. miR-153 has been suggested as a novel tumor-associated miRNA, which is involved in tumor metastasis. However, the clinical significance of miR-153 and its role in PDAC remains to be investigated. The aim of the present study was to investigate the expression levels of miR-153 using RT-qPCR in human PDAC cell lines and tissues. A clinical association analysis was performed to investigate the clinical significance of miR-153. The results showed that, the relative expression of miR-153 in PDAC cells was obviously decreased as compared to that in the normal human pancreatic duct epithelial cell line. The mean expression of miR-153 in PDAC tissues was significantly reduced as compared to that in the normal pancreatic tissues. The clinical analysis revealed that a low expression of miR-153 was closely associated with poor prognostic features and shorter long-term survival of PDAC patients. Furthermore, univariate and multivariate Cox regression analyses showed that miR-153 was an independent prognostic factor for predicting survival in PDAC patients. In vitro studies demonstrated that the upregulation of miR-153 inhibited migration and invasion in MIAPaCa-2 cells. By contrast, the downregulation of miR-153 increased the number of migrated and invaded AsPC-1 cells. miR-153 inversely regulated SNAI1 abundance in MIAPaCa-2 cells. Notably, SNAI1 was identified as a direct target of miR-153 in PDAC. Furthermore, an inverse correlation between miR-153 and SNAI1 expression was observed in PDAC tissues. In conclusion, the results showed miR-153 is an independent prognostic marker for predicting survival in PDAC patients and inhibits cell migration and invasion by targeting SNAI1.

No MeSH data available.


Related in: MedlinePlus

SNAI1 is identified as a functional target of miR-153 in PDAC. (A) RT-qPCR and (B) western blot analysis of SNAI1 expression in MIAPaCa-2 cells with miR-153 or control vector transfection. n, three independent experiments, *P<0.05. (C) miR-153 and its putative binding sequence in the 3′-UTR of SNAI1. The mutant miR-153 binding site was generated in the complementary site for the seed region of miR-153 (wt, wild-type; mt, mutant type). (D) miR-153 significantly suppressed the luciferase activity that carried wt but not mt 3′-UTR of SNAI1. Anti-miR-153 led to a marked increase in luciferase activity of wt 3′-UTR of SNAI1. n, three repeats with similar results, *P<0.05. PDAC, pancreatic ductal adenocarcinoma; 3′-UTR, 3′-untranslated region.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4487667&req=5

f4-or-34-02-0595: SNAI1 is identified as a functional target of miR-153 in PDAC. (A) RT-qPCR and (B) western blot analysis of SNAI1 expression in MIAPaCa-2 cells with miR-153 or control vector transfection. n, three independent experiments, *P<0.05. (C) miR-153 and its putative binding sequence in the 3′-UTR of SNAI1. The mutant miR-153 binding site was generated in the complementary site for the seed region of miR-153 (wt, wild-type; mt, mutant type). (D) miR-153 significantly suppressed the luciferase activity that carried wt but not mt 3′-UTR of SNAI1. Anti-miR-153 led to a marked increase in luciferase activity of wt 3′-UTR of SNAI1. n, three repeats with similar results, *P<0.05. PDAC, pancreatic ductal adenocarcinoma; 3′-UTR, 3′-untranslated region.

Mentions: Additional studies were performed to identify the molecular mechanisms by which miR-153 inhibited cell migration and invasion in PDAC. Previous studies reported that miR-153 was a novel regulator of EMT by targeting SNAI1 and ZEB2 in human epithelial cancers (12). MIAPaCa-2 cells that were transfected with miR-153 and control vectors were subjected to RT-qPCR and western blotting for SNAI1 expression. SNAI1 mRNA and protein levels were significantly reduced by upregulation of miR-153 in MIAPaCa-2 cells (P<0.05, respectively, Fig. 4A and B). To further demonstrate that SNAI1 was directly targeted by miR-153 in PDAC cells, we investigated whether the miR-153 directly interacted with the 3′-UTR of SNAI1 mRNA using a dual-luciferase reporter assay. miR-153 signifi-cantly inhibited the luciferase activity of SNAI1 containing a wt 3′-UTR, but did not suppress the activity of SNAI1 with a mt 3′-UTR (P<0.05, Fig. 4C and D). When anti-miR-153 was transfected, an increase in luciferase activity of wt SNAI1 3′-UTR was observed. However, no relative increase in activity was identified following transfection with the mt SNAI1 3′-UTR constructs (P<0.05, Fig. 4C and D). Thus, our data strongly suggested that SNAI1 is a target of miR-153 in PDAC.


MicroRNA-153 is a prognostic marker and inhibits cell migration and invasion by targeting SNAI1 in human pancreatic ductal adenocarcinoma.

Bai Z, Sun J, Wang X, Wang H, Pei H, Zhang Z - Oncol. Rep. (2015)

SNAI1 is identified as a functional target of miR-153 in PDAC. (A) RT-qPCR and (B) western blot analysis of SNAI1 expression in MIAPaCa-2 cells with miR-153 or control vector transfection. n, three independent experiments, *P<0.05. (C) miR-153 and its putative binding sequence in the 3′-UTR of SNAI1. The mutant miR-153 binding site was generated in the complementary site for the seed region of miR-153 (wt, wild-type; mt, mutant type). (D) miR-153 significantly suppressed the luciferase activity that carried wt but not mt 3′-UTR of SNAI1. Anti-miR-153 led to a marked increase in luciferase activity of wt 3′-UTR of SNAI1. n, three repeats with similar results, *P<0.05. PDAC, pancreatic ductal adenocarcinoma; 3′-UTR, 3′-untranslated region.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4487667&req=5

f4-or-34-02-0595: SNAI1 is identified as a functional target of miR-153 in PDAC. (A) RT-qPCR and (B) western blot analysis of SNAI1 expression in MIAPaCa-2 cells with miR-153 or control vector transfection. n, three independent experiments, *P<0.05. (C) miR-153 and its putative binding sequence in the 3′-UTR of SNAI1. The mutant miR-153 binding site was generated in the complementary site for the seed region of miR-153 (wt, wild-type; mt, mutant type). (D) miR-153 significantly suppressed the luciferase activity that carried wt but not mt 3′-UTR of SNAI1. Anti-miR-153 led to a marked increase in luciferase activity of wt 3′-UTR of SNAI1. n, three repeats with similar results, *P<0.05. PDAC, pancreatic ductal adenocarcinoma; 3′-UTR, 3′-untranslated region.
Mentions: Additional studies were performed to identify the molecular mechanisms by which miR-153 inhibited cell migration and invasion in PDAC. Previous studies reported that miR-153 was a novel regulator of EMT by targeting SNAI1 and ZEB2 in human epithelial cancers (12). MIAPaCa-2 cells that were transfected with miR-153 and control vectors were subjected to RT-qPCR and western blotting for SNAI1 expression. SNAI1 mRNA and protein levels were significantly reduced by upregulation of miR-153 in MIAPaCa-2 cells (P<0.05, respectively, Fig. 4A and B). To further demonstrate that SNAI1 was directly targeted by miR-153 in PDAC cells, we investigated whether the miR-153 directly interacted with the 3′-UTR of SNAI1 mRNA using a dual-luciferase reporter assay. miR-153 signifi-cantly inhibited the luciferase activity of SNAI1 containing a wt 3′-UTR, but did not suppress the activity of SNAI1 with a mt 3′-UTR (P<0.05, Fig. 4C and D). When anti-miR-153 was transfected, an increase in luciferase activity of wt SNAI1 3′-UTR was observed. However, no relative increase in activity was identified following transfection with the mt SNAI1 3′-UTR constructs (P<0.05, Fig. 4C and D). Thus, our data strongly suggested that SNAI1 is a target of miR-153 in PDAC.

Bottom Line: The mean expression of miR-153 in PDAC tissues was significantly reduced as compared to that in the normal pancreatic tissues.Notably, SNAI1 was identified as a direct target of miR-153 in PDAC.In conclusion, the results showed miR-153 is an independent prognostic marker for predicting survival in PDAC patients and inhibits cell migration and invasion by targeting SNAI1.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.

ABSTRACT
Human pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer type with early metastasis, which leads to poor prognosis for patients. Mounting evidence suggests that microRNAs (miRNAs) act as critical factors for tumor recurrence and metastasis. miR-153 has been suggested as a novel tumor-associated miRNA, which is involved in tumor metastasis. However, the clinical significance of miR-153 and its role in PDAC remains to be investigated. The aim of the present study was to investigate the expression levels of miR-153 using RT-qPCR in human PDAC cell lines and tissues. A clinical association analysis was performed to investigate the clinical significance of miR-153. The results showed that, the relative expression of miR-153 in PDAC cells was obviously decreased as compared to that in the normal human pancreatic duct epithelial cell line. The mean expression of miR-153 in PDAC tissues was significantly reduced as compared to that in the normal pancreatic tissues. The clinical analysis revealed that a low expression of miR-153 was closely associated with poor prognostic features and shorter long-term survival of PDAC patients. Furthermore, univariate and multivariate Cox regression analyses showed that miR-153 was an independent prognostic factor for predicting survival in PDAC patients. In vitro studies demonstrated that the upregulation of miR-153 inhibited migration and invasion in MIAPaCa-2 cells. By contrast, the downregulation of miR-153 increased the number of migrated and invaded AsPC-1 cells. miR-153 inversely regulated SNAI1 abundance in MIAPaCa-2 cells. Notably, SNAI1 was identified as a direct target of miR-153 in PDAC. Furthermore, an inverse correlation between miR-153 and SNAI1 expression was observed in PDAC tissues. In conclusion, the results showed miR-153 is an independent prognostic marker for predicting survival in PDAC patients and inhibits cell migration and invasion by targeting SNAI1.

No MeSH data available.


Related in: MedlinePlus