DataSheet_1_Overexpression of Pyruvate Dehydrogenase Kinase-3 Predicts Poor Prognosis in Urothelial Carcinoma.docx

BackgroundThe mitochondrial pyruvate dehydrogenase complex (PDC) link glycolysis to the tricarboxylic acid cycle by decarboxylating pyruvate to acetyl coenzyme A irreversibly. Cancer cells are characterized by a shift in cellular metabolism from mitochondrial respiration to glycolysis. PDC activity inhibition mediated by phosphorylation via pyruvate dehydrogenase kinase (PDK) has been linked to cancer. However, the clinical significance of PDKs in urothelial cancer prognosis is not clear. We investigated the role and prognostic value of PDK3 expression in patients with upper urinary tract urothelial carcinoma (UTUC) and urinary bladder urothelial carcinoma (UBUC).Patients and MethodsWe retrospectively analyzed clinical data and pathological features. Formalin-fixed urothelial carcinoma (UC) tissues were collected and embedded in paraffin. The correlation of PDK3 expression with clinical characteristics, pathological findings and patient outcomes, including metastasis-free survival (MFS) and disease-specific survival (DSS) were analyzed by Pearson’s chi-square test, Kaplan–Meier analysis, and the multivariate Cox proportional hazards model.ResultsData from 295 patients with UBUC and 340 patients with UTUC were evaluated. High PDK3 expression significantly correlated with several pathologic variables such as high T stage, lymph node metastases, high tumor grade, vascular invasion, and high mitotic rate (all P ConclusionHigh PDK3 expression has been linked to negative pathologic characteristics and poor oncological outcomes, suggesting that it could be used as a predictive biomarker for UC. PDK3 mRNA levels and its co-upregulated genes were strongly associated with DNA replication and repair. These results suggest that PDK3 may play a key role in tumor proliferation and development.</p

sj-docx-2-onc-10.1177_11795549221113244 – Supplemental material for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma

Supplemental material, sj-docx-2-onc-10.1177_11795549221113244 for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma by Sung-Wei Lee, Ching-Chieh Yang, Hong-Yue Lai, Hsin-Hwa Tsai, Cheng-Fa Yeh, Yu-Hsuan Kuo, Nai-Wen Kang, Tzu-Ju Chen and Shih-Lun Chang in Clinical Medicine Insights: Oncology</p

sj-docx-3-onc-10.1177_11795549221113244 – Supplemental material for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma

Supplemental material, sj-docx-3-onc-10.1177_11795549221113244 for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma by Sung-Wei Lee, Ching-Chieh Yang, Hong-Yue Lai, Hsin-Hwa Tsai, Cheng-Fa Yeh, Yu-Hsuan Kuo, Nai-Wen Kang, Tzu-Ju Chen and Shih-Lun Chang in Clinical Medicine Insights: Oncology</p

sj-docx-1-onc-10.1177_11795549221113244 – Supplemental material for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma

Supplemental material, sj-docx-1-onc-10.1177_11795549221113244 for Roundabout Guidance Receptor 1 Is an Emerging Prognostic Biomarker for Nasopharyngeal Carcinoma by Sung-Wei Lee, Ching-Chieh Yang, Hong-Yue Lai, Hsin-Hwa Tsai, Cheng-Fa Yeh, Yu-Hsuan Kuo, Nai-Wen Kang, Tzu-Ju Chen and Shih-Lun Chang in Clinical Medicine Insights: Oncology</p

Figure-S4 from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

Figure-S4 AMACR expression does not contribute to tumor angiogenesis. In NMFH-1 (upper) and NMFH-2 (lower) cell lines, the HUVEC tube-forming capability is not significantly attenuated by conditioned media following interference with shAMACR, with similar formation of vascular network as plotted in the histograms.</p

Figure-S7 from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

Figure-S7 Ebselen oxide (EO) at 40 μM significantly increases the subG0-1 cell populations in myxofibrosarcoma cell lines. In the flow cytometric analysis for both NMFH-1 (upper) and NMFH-2 (lower) cell lines, the significantly increased cell percentage in the subG0-1 phase is induced by EO treatment at 40 μM and especially prominent in the NMFH-1. However, neither vehicle controls nor EO treatment at 20 μM shows such an apoptosis-inducing effect.</p

Supplementary Methods and Tables from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

The file contains Supplementary Methods and Tables. TableTable -S1. Clinicopathologic characters of 105 myxofibrosarcoma Table-S2. Univariateand multivariate survivalanalyses for prognostic factors in 89 patients with follow-up Table-S3. Correlations between AMACR and SKP2 immunoexpression TableTable -S4. Results of Cox multivariate analyses in relation to DSS and MeFS in a subset cohort containing informative immunohistochemical data of both AMACR and SKP2 Table-S5Three differentially expressed cell cycle-associatedgenesin AMACR-knockdownmyxofibrosarcoma cell lines Table -S6. Correlations between AMACR, Cyclin D1, Cyclin T2, and Ki-67 immunoexpression</p

Figure-S6 from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

Figure-S6 The in vitro (A) and in vivo (B) downregulating effects of EO treatment on cyclin T2, cyclin D1, and Ki-67 levels. (A): In both NMFH-1 and NMFH-2 myxofibrosarcoma cell lines, western blots (left) reveal reduced expression levels of cyclin T2, cyclin D1, and Ki-67 cell cycle regulatory proteins in cells treated with EO at 40 uM, which, as illustrated in the corresponding histograms (right), are statistically significant in triplicate experiments compared to the control groups. (B) The comparison of immunohistochemical results between EO-treated (right) and control (left) NMFH-2 xenograft specimens shows significantly lower nuclear labeling indices of cyclin T2, cyclin D1, and Ki-67.</p

Figure-S3 from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

Figure-S3 AMACR expression has no effect on migration/invasion of myxofibrosarcoma cells. In NMFH-1 (upper) and NMFH-2 (lower) cell lines with shAMACR, the capabilities of wound closure (A) and cell invasiveness (B) are not significantly impaired, compared with controls. Experiments are performed in triplicate and results expressed as mean {plus minus} SD.</p

Figure-S2 from <i>AMACR</i> Amplification in Myxofibrosarcomas: A Mechanism of Overexpression That Promotes Cell Proliferation with Therapeutic Relevance

Figure-S2 Apoptosis not induced in AMACR-knockdown myxofibrosarcoma cells. In both NMFH-1 (upper) and NMFH-2 (lower) cell lines, the cell percentages in the early and late stages of apoptosis analyzed by annexin V//propodium iodine staining are not significantly different between shLacZ controls and shAMACR-treated cells in three independent assays.</p