Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: Correlation between miR-23a and FOXM1 expression in human pancreatic ductal adenocarcinoma (PDAC) tissues. (A) Total mRNA including miRNA was extracted from distinct cohorts of pancreatic tumor samples with varied levels of FOXM1 expression (negative/weak, moderate, and strong) through laser-assisted microdissection ( n = 10). (B, C) The miR-23a expression levels were quantified using quantitative PCR in triplicate. Notably, an inverse correlation was observed between FOXM1 protein expression and miR-23a-3p ( n = 30; r = −0.5400, P = 0.0021) and -5p ( n = 30; r = −0.6374, P = 0002) expression. (D) Decreased level of both miR-23a-3p and -5p was apparent in human pancreatic cancer cell lines. (E, F) The levels of miR-23a-3p and -5p in high FOXM1 mRNA expressing human pancreatic cells were lower than those in low FOXM1 mRNA expressing cells. An inverse correlation was also observed between FOXM1 mRNA expression and miR-23a-3p expression.

Article Snippet: All the miRNAs were reverse transcribed into the first strand of cDNA by A-tailing using miRcute Plus miRNA First-Strand cDNA Kit (TIANGEN, CAT# KR211-02), and were further amplified using miRcute Plus miRNA qPCR Kit (SYBR Green) (TIANGEN, CAT# FP411-02) with designed forward primer of miRNAs and universal reverse primer in the kit.

Techniques: Expressing, Laser Capture Microdissection, Real-time Polymerase Chain Reaction

Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: Decreased miR-23a expression in three different mouse models of ADM in vivo . (A, B) In vivo ADM induction using three mouse models generated as described in the Materials and Methods section, including PDL, CAE, and KC mouse models of pancreatic cancer. FoxM1 and miR-23a expression were determined with quantitative PCR in triplicate. The induction of ADM was accompanied by decreased expression of both miR-23a-3p and miR-23a-5p in KC, PDL, and CAE mouse models ( n = 5). Note that the increased expression of FoxM1 correlated with a significantly decreased expression of miR-23a-3p and miR-23a-5p. (C) Mice were treated with caerulein for two days and miR-23a expression was determined with quantitative PCR 2, 5, and 8 days after the treatment. Caerulein treatment decreased the expression of both miR-23a-3p and miR-23a-5p, while the decreased level of miR-23a-3p and -5p recovered after caerulein treatment was stopped for eight days. PDL, pancreatic ductal ligation; CAE, caerulein treatment; KC, Pdx1-Cre; LSL-Kras G12D/+ . ADM, acinar-to-ductal metaplasia.

Article Snippet: All the miRNAs were reverse transcribed into the first strand of cDNA by A-tailing using miRcute Plus miRNA First-Strand cDNA Kit (TIANGEN, CAT# KR211-02), and were further amplified using miRcute Plus miRNA qPCR Kit (SYBR Green) (TIANGEN, CAT# FP411-02) with designed forward primer of miRNAs and universal reverse primer in the kit.

Techniques: Expressing, In Vivo, Generated, Real-time Polymerase Chain Reaction, Ligation

Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: Induction of ADM decreased the expression of miR-23a in primary acinar cells and acinar cells in vitro . (A) Primary acinar cells were treated with TGF-α for three days, immunofluorescence staining was performed using specific antibodies against CK19 and amylase. Note that the induction of ADM was accompanied by an increased level of CK19 and decreased expression of amylase. (B) Primary acinar cells, MPC-83, and 266-6 cells were treated with or without 50 ng/mL TGF-α for three days. miR-23a-3p and miR-23a-5p levels were determined with quantitative PCR in triplicate. Note that induction of ADM correlated with a significantly decreased expression of miR-23a-3p and miR-23a-5p in primary acinar cells and MPC-83 and 266-6 pancreatic cells. ADM, acinar-to-ductal metaplasia.

Article Snippet: All the miRNAs were reverse transcribed into the first strand of cDNA by A-tailing using miRcute Plus miRNA First-Strand cDNA Kit (TIANGEN, CAT# KR211-02), and were further amplified using miRcute Plus miRNA qPCR Kit (SYBR Green) (TIANGEN, CAT# FP411-02) with designed forward primer of miRNAs and universal reverse primer in the kit.

Techniques: Expressing, In Vitro, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: miR-23a down-regulated the expression of FOXM1 in both mouse pancreatic cells and human pancreatic cancer cells. (A – C) CFPAC-1 and MIA PaCa-2 human PDAC cells and MPC-83 mouse pancreatic cells were treated with either miR-23a-3p or miR-23a-5p for 48 h, and FOXM1 protein and mRNA expression were determined with western blot analyses and quantitative PCR in triplicate. (D – F) CFPAC-1 and MIA PaCa-2 human PDAC cells and MPC-83 mouse pancreatic cells were treated with inhibitors of either miR-23a-3p or miR-23a-5p for 48 h and FOXM1 protein and mRNA expression were determined with western blot analyses and quantitative PCR. Note that miR-23a-3p and miR-23a-5p inhibited the expression of FOXM1 protein and mRNA in CFPAC-1 and MIA PaCa-2 human pancreatic cancer cells and MPC-83 mouse pancreatic cells, while the inhibitors of miR-23a-3p and miR-23a-5p increased the expression of FOXM1 protein and mRNA in CFPAC-1 and MIA PaCa-2 human pancreatic cancer cells and MPC-83 mouse pancreatic cell.

Article Snippet: All the miRNAs were reverse transcribed into the first strand of cDNA by A-tailing using miRcute Plus miRNA First-Strand cDNA Kit (TIANGEN, CAT# KR211-02), and were further amplified using miRcute Plus miRNA qPCR Kit (SYBR Green) (TIANGEN, CAT# FP411-02) with designed forward primer of miRNAs and universal reverse primer in the kit.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction