Fig 1: The generation and characterization of JCPyV T antigen transgenic mice. CAG-loxp-Laz-loxp-T antigen transgenic mice were established using a CAG promoter as outlined in the schematic diagram. To activate tissue-specific expression of T antigen, the mice were crossed with Atp4b-cre (gastric parietal cells), PGC-cre (gastric chief cells), Capn8-cre (gastric pit cells), K19-cre (stem-like cells), villin-cre (intestinal epithelial cells), Alb-cre (hepatocytes), and Pdx1-cre (pancreas and gastrointestinal cells) mice (A). These positive mice showed cre + /T antigen + after the PCR of tail DNA (B). To verify the successful knockout of LaZ, we performed PCR on the DNA of promoter-specific organs targeting the LacZ and CAG/T antigen (C). T-antigen expression was examined in target organs of transgenic mice by western blot (D). Finally, RT-PCR was employed to detect alternative splicing of the T antigen intron using the mRNA of target organs in transgenic mice (E). AlJ, Alb-cre/ JCPyV T antigen; CaJ, Capn8-cre/JCPyV T antigen; IRES, internal ribosome entry site; ITR, inverted terminal repeat; K19J, K19-cre/JCPyV T antigen; NC, negative control; PdJ, Pdx1-cre/JCPyV T antigen; PGJ, PGC-cre/JCPyV T antigen; PvJ, villin-cre/CPyV T antigen; RT-PCR, reverse transcription PCR; T28, PBS-T antigen plasmid as positive control; WT, wild-type; 4bJ, Atp4b-cre/JCPyVT antigen
Fig 2: Multiple tumorigeneses were found in PGC-cre/JCPyV T antigen mice. Gastric cancer (A, ♀, 12 months; B, ♂,17 months) was histologically found in the PGC- cre/JCPyV T antigen transgenic mouse. Colorectal well-differentiated adenocarcinoma (C, ♂, 17 months) was also seen in transgenic mouse of PGC-cre/JCPyV T antigen. Breast tumor was grossly observed in the belly of the PGC-cre/JCPyV T antigen transgenic mouse (♀ 12 months, D). After HE staining (E) and immunostaining (F), lobular carcinoma of the breast was found to express T antigen, GATA-3, CA153, and β-catenin. The expression of PGC was immunohistochemically examined in gastric and breast cancers, and matched normal glands (G) as outlined in the table (H). The TCGA database was employed to analyze PGC mRNA expression in gastric and breast cancers, and their corresponding normal tissues (I). Finally, PGC content was determined by ELISA or western blot in the serum of a healthy individual, atrophic gastritis, gastric cancer, and human and bovine milks (J). AG, atrophic gastritis; BC, breast cancer; BN, breast normal tissue; CA153, carbohydrate antigen 153; ELISA, enzyme-linked immunosorbent assay; ER, estrogen receptor; GC, gastric cancer; GN, gastric normal tissue; HE, hematoxylin and eosin; HI, healthy individual; HM, human milk; M, milk powder; PC, positive control; PGC, pepsinogen C; PR, progesterone receptor; TCGA, The Cancer Genome Atlas
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