FIGURE 2 from Metronomic Administration of Topotecan Alone and in Combination with Docetaxel Inhibits Epithelial–mesenchymal Transition in Aggressive Variant Prostate Cancers
Authors: Mitra Ghosh Taraswi , Mazumder Suman , Davis Joshua , Yadav Jyoti , Akinpelu Ayuba , Alnaim Ahmed , Kumar Harish , Waliagha Razan , Church Bird Allison E. , Rais-Bahrami Soroush , Bird R. Curtis , Mistriotis Panagiotis , Mishra Amarjit , Yates Clayton C. , Mitra Amit K. , Arnold Robert D. 
Published: July 2023

<p><i>In silico and in vitro</i> validation of top EMT marker's association with disease progression, patient survival, and disease aggressiveness. <b>A,</b><i>In silico</i> correlation of EMT signatures using TCGA database: Top EMT marker <i>TGFB1</i> and taxane resistance marker <i>FSCN1</i> were associated with DFS of patients with prostate cancer. (<b>I</b>) High expression of <i>FSCN1</i> was correlated with lower survival compared with the low expression group of patients (<i>P</i> < 0.0001). (<b>II</b>) Higher expression of <i>TGFB1</i> showed less survival compared with the low expression group of patients (<i>P</i> < 0.0001). <b>B,</b> Assessment of “stem-like” cells (CD44<sup>high</sup>/CD133<sup>high</sup>) population in prostate cancer subtypes by Flowcytometry: Assessing of “stem-like” cell population among all prostate cancer subtypes. (<b>I</b>) Baseline (no drug treatment) CD44<sup>+</sup> cells were higher in EA-AR<sup>Low</sup>/mCRPC (PC-3-76.5%, PC-3M-90.0%, DU145-69.0%) compared with EA-AR<sup>High</sup>/mCSPC (LNCaP-5.60%, VCaP-6.69%, 22RV1-4.51%). Taxane-resistant AR<sup>Low</sup>/mCRPC (DUTXR) cells showed the highest percent of “stem-like” cells (CD44<sup>+</sup> cells 96.4%). Furthermore, AA-AR<sup>High</sup> /mCSPC (MDA-PCa-2b) showed higher percent of CD44<sup>+</sup> (16.6%) cells compared with EA-AR<sup>High</sup>/mCSPC (LNCaP, VCaP, 22RV1). Whereas in contrast, another AA-AR<sup>High</sup>/mCSPC (RC77T/E) showed low percent CD44<sup>+</sup> cells population (5.69%). EA-AR<sup>Present</sup>/mCRPC (C4-2B) showed CD44<sup>+</sup> cell population (8.50%). (<b>II</b>) and (<b>III</b>) Bar graphs represented a comparison among CD44<sup>+</sup> and CD44<sup>+</sup>/CD133<sup>+</sup> cells in all prostate cancer subtypes. (*, <i>P</i> ≤ 0.05). Results represented higher stemlike cell population in AR<sup>Low</sup>/mCRPC and taxane-resistant mCRPC. <b>C,</b> Immunoblotting: Higher expression of EMT proteins (CD44, ALDH1, Oct-4, TGFβ, Nanog, and Sox2) was observed in AR<sup>Low</sup>/mCRPC/NEPC (PC-3, PC-3M, DU145) prostate cancer cell lines compared with AR<sup>High</sup>/mCSPC (LNCaP, VCaP, 22RV1) cell lines. Furthermore, higher expression of EMT proteins (CD44, ALDH1, Oct-4, TGFβ, Nanog, and Sox2) was observed in AA-AR<sup>High</sup>/mCSPC (MDA-PCa-2b, RC77T/E) compared with EA-AR<sup>High</sup>/mCSPC (LNCaP, VCaP, 22RV1) prostate cancer cell lines. Highest amount of EMT protein was expressed in taxane-resistant (DUTXR and PC-3TXR) prostate cancer cell lines. Immunoblotting results corroborated with DEGs. Densitometry plots represented a comparison of protein expression among all prostate cancer subtypes. Actin-β was used as a control housekeeping gene to normalize the protein expression of other genes for immunoblotting experiments (*, <i>P</i> ≤ 0.05).</p>