Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: Relative N6-methyladenine (m6A) RNA modification in total RNA is upregulated in pancreatic cancer (PC) (A and B) m6A content in total RNA in 16 pairs of pancreatic adenocarcinoma (PAAD) and corresponding para-tumor tissues. Except for patients 3 and 8, the relative m6A RNA modification of PAAD tissues was significantly higher than that of the corresponding para-tumor tissues. (C) Relative m6A RNA modification was higher in PC cell lines. Human PC cell lines (SW1990, AsPC-1, PANC-1, BxPC-3, and Capan-2) and a human immortalized pancreatic duct epithelial cell line (HPDE6-C7) were subjected to RNA m6A detection. Tumor, PAAD tissue. (B) ∗p < 0.05 by paired Student’s t test; (C) ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by unpaired Student’s t test. Data are represented as mean ± standard error.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: RNA modification

Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: FTO acts as m6A demethylase in pancreatic cell lines (A and B) Quantitative real-time PCR and WB assays showed reduced FTO levels in five PC cell lines. The mRNA and protein levels of FTO were (C and E) significantly reduced in SW1990 cells after small interfering (si)RNAs (siFTO-1, siFTO-2, and siFTO-3) transfection, whereas were (D and F) significantly increased in BxPC-3 cells after transfection with wild-type FTO (FTO-WT) and mutant FTO (FTO-mut) overexpressing plasmids. (G) FTO knockdown significantly upregulated the m6A level in total RNA of SW1990 cells. (H) FTO-WT overexpression significantly decreased the m6A level in total RNA of BXPC-3 cells. No significant changes were observed in the m6A levels upon FTO-mut overexpression. GAPDH was used as internal control in the quantitative real-time PCR and WB assays. NC, negative control. (A–H) ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ns, no statistical significance by unpaired Student’s t test.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Transfection, Mutagenesis, Knockdown, Over Expression, Control, Negative Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: m6A demethylase FTO suppresses pancreatic cancer tumorigenesis by demethylating PJA2 and inhibiting Wnt signaling

doi: 10.1016/j.omtn.2021.06.005

Figure Lengend Snippet: FTO enhanced PJA2 stability in an m6A-YTHDF2-dependent manner in PC cells (A and B) Venn diagrams illustrated overlap in differentially bioinformatics analyses of the top 20 genes significantly positively or negatively related to FTO in PC. (C–F) Quantitative real-time PCR was performed to detect the mRNA expression of eight candidate genes after knockdown and overexpression of FTO in BxPC-3 and SW1990 cells. Only the PJA2 expression pattern, by both quantitative real-time PCR and WB, was consistent with that of FTO. (G) cBioPortal analysis displayed the co-expression relationship between FTO and PJA2 in PAAD tissues. Spearman coefficient: 0.74 (p = 2.10e−32); Pearson coefficient: 0.75 (p = 7.98e−34). (H and I) The relative PJA2 m6A level was detected in BxPC-3 and SW1990 cells after knockdown and overexpression of FTO, respectively. (J–M) Quantitative real-time PCR and WB were performed to detect the mRNA and protein expression of PJA2 after YTHDF1 and YTHDF2 knockdown by siRNA in BxPC-3 and SW1990 cells. (N and O) PJA2 stability was detected after knocking down YTHDF2, FTO by siRNA, and overexpression of FTO-WT and FTO-mut in BxPC-3 and SW1990 cells. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

Article Snippet: Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCC CRL-1682), PANC-1 (ATCC CRL-1469), BxPC-3 (ATCC CRL-1687), and Capan-2 (ATCC HTB-80) were purchased from ATCC (Manassas, VA, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Knockdown, Over Expression