Figure 2.
SB mutagenesis in NPp53 mice promotes NEPC. (A) Strategy: NPp53-SB(−) and NPp53-SB(+) mice (2–3 mo of age) were induced to form tumors by delivery of tamoxifen and the mice were monitored for up to 15 mo, following which tissues were collected for phenotypic (i.e., histological), transcriptomic (i.e., RNA-seq), and genomic (i.e., CIS) analyses. (B) Kaplan–Meier survival analysis of NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 93) mice. The P value was calculated using a Log-rank test. (C) Metastasis frequency in NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 84) mice comparing non-NEPC (n = 52) and NEPC (n = 32) tumors. The P value was calculated using the Fisher’s exact test. (D and E) Distribution of histological variant phenotypes and metastasis incidence in NPp53-SB(−) (n = 30) and NPp53-SB(+) (n = 67) mice. Histological phenotypes were classified as mouse prostatic intraepithelial neoplasia (mPIN), adenocarcinoma (ADC), squamous (Squam), neuroendocrine (NE), sarcomatoid (Sarcom), or undifferentiated (Undif). (D) Distribution of primary histology (pie chart) and secondary histology (donut chart). (E) Distribution of primary histology (pie chart) and metastatic status (donut chart). (F) NEPC frequency in NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 84) prostate tumors. The P value was calculated using the Fisher’s exact test. (G and H) Histopathological analyses. Representative images of prostate tumors from NPp53-SB(−) and NPp53-SB(+) mice, showing examples of NEPC and non-NEPC (adenocarcinoma and sarcomatoid) histopathology. Shown are H&E (G), and immunohistochemical (IHC; G) or immunofluorescence (IF; H) staining for markers of prostate cancer and NEPC, namely, AR, synaptophysin (Syp), Pan-cytokeratin (Pan-CK), vimentin (Vim), Ki67, and Sox2. Analyses were done on a minimum of five independent tumors for each group and each condition. Scale bars represent 20 µm for H&E and IHC, 10 µm for IF images. (I) Quantitative real-time PCR showing expression levels of AR, Syp, and chromogranin A (ChgA) in non-NEPC (n = 6) and NEPC (n = 7) NPp53-SB(+) tumors. Data are normalized using Gapdh as internal control. Analyses were done on six to seven independent samples with a minimum of two biological replicates. See also Fig. S1, Table 1, and Table S1. Refer to the image caption for details. Panel A shows a schematic diagram of the experimental strategy involving NPp53-SB negative and NPp53-SB positive mice. Panel B shows a Kaplan–Meier survival analysis graph with the y-axis labeled Survival probability percentage and the x-axis labeled Days after TAM, comparing NPp53-SB negative and NPp53-SB positive mice. Panel C shows a bar graph of metastasis frequency with the y-axis labeled Metastasis frequency percentage and groups labeled Non-NEPC and NEPC for both NPp53-SB negative and NPp53-SB positive mice. Panel D consists of pie charts and donut charts showing the distribution of histological variant phenotypes and metastasis incidence in NPp53-SB negative and NPp53-SB positive mice. Panel E consists of pie charts and donut charts showing the distribution of primary histology and metastatic status in NPp53-SB negative and NPp53-SB positive mice. Panel F shows a bar graph of NEPC frequency with the y-axis labeled NEPC frequency percentage, comparing NPp53-SB negative and NPp53-SB positive prostate tumors. Panel G shows representative images of prostate tumors with hematoxylin and eosin staining and immunohistochemical staining for various markers. Panel H shows representative images of prostate tumors with immunofluorescence staining for various markers. Panel I consists of bar graphs showing the expression levels of androgen receptor, synaptophysin, and chromogranin A in non-NEPC and NEPC NPp53-SB positive tumors.

SB mutagenesis in NPp53 mice promotes NEPC. (A) Strategy: NPp53-SB(−) and NPp53-SB(+) mice (2–3 mo of age) were induced to form tumors by delivery of tamoxifen and the mice were monitored for up to 15 mo, following which tissues were collected for phenotypic (i.e., histological), transcriptomic (i.e., RNA-seq), and genomic (i.e., CIS) analyses. (B) Kaplan–Meier survival analysis of NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 93) mice. The P value was calculated using a Log-rank test. (C) Metastasis frequency in NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 84) mice comparing non-NEPC (n = 52) and NEPC (n = 32) tumors. The P value was calculated using the Fisher’s exact test. (D and E) Distribution of histological variant phenotypes and metastasis incidence in NPp53-SB(−) (n = 30) and NPp53-SB(+) (n = 67) mice. Histological phenotypes were classified as mouse prostatic intraepithelial neoplasia (mPIN), adenocarcinoma (ADC), squamous (Squam), neuroendocrine (NE), sarcomatoid (Sarcom), or undifferentiated (Undif). (D) Distribution of primary histology (pie chart) and secondary histology (donut chart). (E) Distribution of primary histology (pie chart) and metastatic status (donut chart). (F) NEPC frequency in NPp53-SB(−) (n = 31) and NPp53-SB(+) (n = 84) prostate tumors. The P value was calculated using the Fisher’s exact test. (G and H) Histopathological analyses. Representative images of prostate tumors from NPp53-SB(−) and NPp53-SB(+) mice, showing examples of NEPC and non-NEPC (adenocarcinoma and sarcomatoid) histopathology. Shown are H&E (G), and immunohistochemical (IHC; G) or immunofluorescence (IF; H) staining for markers of prostate cancer and NEPC, namely, AR, synaptophysin (Syp), Pan-cytokeratin (Pan-CK), vimentin (Vim), Ki67, and Sox2. Analyses were done on a minimum of five independent tumors for each group and each condition. Scale bars represent 20 µm for H&E and IHC, 10 µm for IF images. (I) Quantitative real-time PCR showing expression levels of AR, Syp, and chromogranin A (ChgA) in non-NEPC (n = 6) and NEPC (n = 7) NPp53-SB(+) tumors. Data are normalized using Gapdh as internal control. Analyses were done on six to seven independent samples with a minimum of two biological replicates. See also Fig. S1, Table 1, and Table S1.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal