cell culture human pancreatic cancer cell line sw1990  (ATCC)

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: Figure 1. 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: CT A Cell lines, reagents, and culture conditions Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCCCRL-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:

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: Figure 3. 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: CT A Cell lines, reagents, and culture conditions Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCCCRL-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: Figure 6. 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 = 2.10e32); Pearson coefficient: 0.75 (p = 7.98e34). (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: CT A Cell lines, reagents, and culture conditions Human pancreatic cancer cell lines SW1990 (American Type Culture Collection [ATCC] CRL-2172), AsPC-1 (ATCCCRL-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

Journal: Cancer Science

Article Title: Upregulation of transgelin is an independent factor predictive of poor prognosis in patients with advanced pancreatic cancer

doi: 10.1111/cas.12107

Figure Lengend Snippet: Effects of Transgelin on tumorigenesis and tumor growth in vivo. Xenografted pancreatic cancers were established by s.c. injection of SW1990, SW1990/shControl and SW1990/shTransgelin (5 × 106) into 4‐week‐old Balb/c nude mice. Seven days later, they were administered either PBS or gemcitabine (GEM, 50 mg/kg) i.p., twice a week, respectively. (a) Tumor size was measured weekly. (b) Tumor weights were measured at day 35. (c) Immunohistochemical analysis of tumor sections from each treatment group, with antibodies against transgelin and the proliferation marker proliferating cell nuclear antigen (PCNA). *P < 0.05 and **P < 0.01 versus controls.

Article Snippet: Cell lines and cultures Human pancreatic ductal cancer cell lines SW1990 and BxPC3 were originally obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: In Vivo, Injection, Immunohistochemical staining, Marker

Journal: Cancer Science

Article Title: Upregulation of transgelin is an independent factor predictive of poor prognosis in patients with advanced pancreatic cancer

doi: 10.1111/cas.12107

Figure Lengend Snippet: Effects of transgelin on the growth and tumorigenesis of pancreatic cancer cells in vitro. (a) The expression of transgelin in SW1990 and BxPC3 cells stably transfected with pGCsi‐H1‐Transgelin plasmid (SW1990/shTrans‐1, SW1990/shTrans‐2 and BxPC3/shTrans‐1, BxPC3/shTrans‐2) were detected by western blot analysis. Untreated cells and cells transfected with pGCsi‐H1/negative plasmid (SW1990/shControl, BxPC3/shControl) served as the controls. (b) BxPC3/shTransgelin, SW1990/shTransgelin cells and their controls were cultured for 24, 48 and 72 h. WST‐8 cleavage in each group was measured (absorbance OD450). (c) Colony formation assay was performed using SW1990 and BxPC3 cells stably transfected with shTransgelin and shControl for 14 days. (d) Colony formation rates were analyzed by soft agar assay. All experiments were repeated in triplicate with similar results. *P < 0.05 and **P < 0.01 when compared to controls.

Article Snippet: Cell lines and cultures Human pancreatic ductal cancer cell lines SW1990 and BxPC3 were originally obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: In Vitro, Expressing, Stable Transfection, Transfection, Plasmid Preparation, Western Blot, Cell Culture, Colony Assay, Soft Agar Assay

Journal: Cancer Science

Article Title: Upregulation of transgelin is an independent factor predictive of poor prognosis in patients with advanced pancreatic cancer

doi: 10.1111/cas.12107

Figure Lengend Snippet: Effects of Transgelin on the migration and invasion of pancreatic cancer cells. SW1990 and BxPC3 cells stably transfected with pGCsi‐H1‐Transgelin (SW1990/shTrans‐1, SW1990/shTrans‐2 and BxPC3/shTrans‐1, BxPC3/shTrans‐2) or control plasmids (BxPC3/shControl, SW1990/shControl) were subjected to migration, invasion and wound healing assays. Number of cells that migrated or invaded to the lower membrane surface was counted after 12 and 24 h. (a) Cell migration assay was performed using transwell chambers. Representative images are shown for SW1990. (b) Effects of transgelin on cell motility of SW1990 and BxPC3 at 24 h. (c) Invasive potential of cells across Matrigel‐coated transwell chambers. Representative images are shown for SW1990. (d) Effects of transgelin on cell invasion of SW1990 and BxPC3 at 24 h. (e) Representative images at 0, 24 and 48 h after wound initiation are shown for SW1990. (f) Migration index was assessed by measuring the distance between the wound edges for SW1990 and BxPC3 at 48 h. All experiments were repeated in triplicate with similar results. *P < 0.05 and **P < 0.01 versus controls.

Article Snippet: Cell lines and cultures Human pancreatic ductal cancer cell lines SW1990 and BxPC3 were originally obtained from the American Type Culture Collection (Rockville, MD, USA).

Techniques: Migration, Stable Transfection, Transfection, Control, Membrane, Cell Migration Assay