Journal: Cancer Science

Article Title: SNAIL2 contributes to tumorigenicity and chemotherapy resistance in pancreatic cancer by regulating IGFBP2

doi: 10.1111/cas.15162

Figure Lengend Snippet: SNAIL2 is highly expressed in tumorigenic pancreatic cancer cell lines. A, Microscopy images of 2D cultures and spheres (3D culture, day 7) in KLM1 and KMP5. Scale bars: 100 µm. B, qRT‐PCR (EMT‐TF) of KLM1 and KMP5. Relative expression levels normalized by HPDE cells were compared. n = 3, each. ND, not detected. Mean + SE. ** P < .01. C, qRT‐PCR ( SNAI2 ) of control vector versus shSNAI2 from KLM1 and KMP5. n = 3, each. Mean + SE. ** P < .01. D, Western blot (SNAIL2, CDH1, VIM, and ACTB) of control vector versus shSNAI2 from KLM1 and KMP5. The relative intensity (ratio) is shown below each band. shRNA was used shSNAI2_1

Article Snippet: The SNAIL2 and IGFBP2 overexpression vectors and control vector were purchased from Sino Biologicals.

Techniques: Microscopy, Quantitative RT-PCR, Expressing, Plasmid Preparation, Western Blot, shRNA

Journal: Cancer Science

Article Title: SNAIL2 contributes to tumorigenicity and chemotherapy resistance in pancreatic cancer by regulating IGFBP2

doi: 10.1111/cas.15162

Figure Lengend Snippet: SNAI2 is highly expressed in spheroids established from surgically resected human pancreatic cancer. A, Immunohistochemical SNAIL2 staining of human pancreatic cancer (2 samples). Scale bars: 100 µm. B, Microscope image of spheroids in 3D culture (left), H&E staining of spheroids (middle), H&E staining of primary tumor (right). Scale bars: 100 µm. C, qRT‐PCR (EMT‐TF) of Sphs. Relative expression levels normalized by HPDE cells were compared. n = 3. ND, not detected. Mean + SE. ** P < .01. D, Microscopic image of control vector and shSNAI2 from Sphs in 3D culture. Scale bars: 100 µm. E, qRT‐PCR ( SNAI2 ) of control vector versus shSNAI2 from spheroids. n = 3, each. Mean + SE. * P < .05, ** P < .01. F, Western blot (SNAIL2, CDH1, VIM, and ACTB) of control vector versus shSNAI2 from Sph. shRNA was used shSNAI2_1. The relative intensity (ratio) is shown below each band

Article Snippet: The SNAIL2 and IGFBP2 overexpression vectors and control vector were purchased from Sino Biologicals.

Techniques: Immunohistochemical staining, Staining, Microscopy, Quantitative RT-PCR, Expressing, Plasmid Preparation, Western Blot, shRNA

Journal: Cancer Science

Article Title: SNAIL2 contributes to tumorigenicity and chemotherapy resistance in pancreatic cancer by regulating IGFBP2

doi: 10.1111/cas.15162

Figure Lengend Snippet: Gene transduction of IGFBP2 and SNAI2 into shSNAI2 restores mRNA and protein expression and improves tumorigenicity and resistance to gemcitabine. A, qRT‐PCR ( SNAI2 and IGFBP2 ) in sh+Vector, sh+IGFBP2 and sh+SNAI2 from KLM1. n = 3, each. Mean + SE. ** P < .01. B, Western blot (SNAIL2, IGFBP2, and ACTB) of sh+Vector versus sh+IGFBP2 and sh+SNAI2 from KLM1 cells. The relative intensity (ratio) is shown below each band. C, D, Sphere formation assay of parental cells, sh+Vector, sh+IGFBP2 and sh+SNAI2 from KLM1 cell at day 7. C, Microscope images. Scale bars: 100 µm. D, Quantification of the number of spheres. n = 3, each. Mean + SE. ** P < .01. E‐H, sh+Vector, sh+IGFBP2, and sh+SNAI2 cells from KLM1 cells were subcutaneously transplanted into NOD‐SCID mice (10 5 cells, 10 4 cells per site), respectively. n = 3. E, Macroscope images of the formed tumor. F, Number of tumor‐forming mice per total transplanted mice at 12 wk after transplantation. G, Tumor growth curves. Mean + SE. * P < .05, ** P < .01. H, H&E staining of the formed tumor. The mesenchymal area (indicated by arrows) ratio is shown below each picture. Scale bars: 100 µm. I, The relative viability of parent cells, sh+Vector versus sh+IGFBP2, and sh+SNAI2 from KLM1 cells 96 h after treatment with gemcitabine. Viability curves at 0, 1, 10, 100 ng/mL gemcitabine. n = 4, each. Mean + SE. ** P < .01

Article Snippet: The SNAIL2 and IGFBP2 overexpression vectors and control vector were purchased from Sino Biologicals.

Techniques: Transduction, Expressing, Quantitative RT-PCR, Plasmid Preparation, Western Blot, Tube Formation Assay, Microscopy, Transplantation Assay, Staining