Further Understanding of Urokinase Plasminogen Activator Overexpression in Urothelial Bladder Cancer Progression, Clinical Outcomes and Potential Therapeutic Targets

Figures & data

Table 1 Overview of the Studies Investigating the Plasminogen Activation System (PAS) in vitro

In vitro Studies
Author	Year	Pat. No.	Study Design	Marker Investigated	Cell Line/Source	Findings
McGarvey et al^Citation25	1998	6 cell lines, 29 tissue specimens	retrospective	uPA, uPAR, PAI-1	UBC cell lines (RT4, HT-5637, HT-1376, TCCsup, J82, T24), tissue	uPA, uPAR, PAI-1 overexpression in UBC cell lines is associated with higher tumor stage (p<0.05) uPA overexpression in tissue is associated with higher tumor stage (p<0.003) uPAR overexpression in tissue is associated with higher tumor stage (p<0.008)
Champelovier et al^Citation26	2002	Preliminary experiment: 7 cell lines, 15 tissue specimens Validation: 129 tissue specimens	retrospective	uPA, tPA, uPAR, PAI-1, PAI-2	Exploratory analysis: UBC cell lines (RT4, RT112, CHA 89, T24, J82S, DAG-1, TCCsup), tissue Validation cohort: tissue	Exploratory analysis: uPAR overexpression in UBC cell lines is associated with higher tumor grade (p=0.02) and cell motility (p=0.014) tPA overexpression in tissue is associated with higher tumor grade (p=0.05) uPAR overexpression in tissue is associated with higher tumor stage (p=0.02) and higher tumor ploidy (p=0.04) PAI-2 overexpression in tissue is associated with higher tumor stage (p=0.02), higher tumor grade (p=0.03) and higher tumor ploidy (p=0.04) Validation cohort: uPAR overexpression is associated with higher tumor grade (p=0.002), higher tumor stage (p=0.003), higher tumor ploidy (p=0.05) and shorter OS (p=0.045) PAI-2 overexpression is associated with higher tumor grade (p=0.005) and shorter OS (p=0.038)
Hau et al^Citation24	2017	5 cell lines	not reported	uPAR	UBC cell lines (UROtsa, RT4, UM-UC-3, T24, J82)	uPAR overexpression is associated with higher tumor stage (p<0.05) uPAR gene-silencing inhibited cell migration >60% (p<0.005) and cell invasion >80% (p<0.005)

Abbreviations: UBC, urinary bladder carcinoma; uPA, urokinase-type plasminogen activator; tPA, tissue-type plasminogen activator; uPAR, uPA receptor; PAI-1, plasminogen activator inhibitor 1; PAI-2, plasminogen activator inhibitor 2; OS, overall survival.

Table 2 Overview of the Studies Investigating the Plasminogen Activation System (PAS) on Non-Muscle-Invasive Bladder Cancer (NMIBC)

NMIBC Tissue
Author	Year	Pat. No.	Study Design	Marker Investigated	Source	Findings
Hasui et al^Citation31	1994	42	not reported	uPA	Tissue	uPA overexpression is associated with shorter PFS (p<0.005)
Hasui et al^Citation32	1996	52	prospective	uPA	Tissue	uPA overexpression is associated with shorter OS (p<0.005) and is an independent predictor for worse OS (RR 6.22; p=0.020) and shorter PFS (RR 5.93; p=0.011) Muscle invasion occurred in 2 (12%) patients in the uPA overexpression group versus 1 (3%) in the low-expression group (p-value not shown)
Iwata et al^Citation28	2019	827	retrospective	uPA, uPAR, PAI-1	Tissue	uPA overexpression is associated with higher tumor grade (p=0.016) and is an independent predictor for shorter RFS (HR 1.4, 95% CI: 1.10–1.78; p=0.006) and shorter PFS (HR 1.68, 95% CI: 1.04–2.74; p=0.035) in all NMIBC tumors and shorter RFS (HR 1.64, 95% CI: 1.07–2.51; p=0.022) and shorter PFS (HR 2.19, 95% CI: 1.15–4.18; p=0.018) in T1G3 tumors uPAR overexpression is associated with higher tumor grade (p=0.004) and is an independent predictor for shorter RFS (HR 1.7, 95% CI: 1.33–2.17; p<0.001) in all NMIBC tumors and shorter RFS in TaG1-2 (HR 1.54, 95% CI: 1.13–2.09; p=0.006) and T1G3 tumors (HR 2.0, 95% CI: 1.29–3.12; p=0.002) PAI-1 overexpression is associated with higher tumor grade (p=0.001) and is an independent predictor for shorter RFS (HR 1.35, 95% CI: 1.06–1.71; p=0.014) in all NMIBC tumors and shorter RFS (HR 1.46, 95% CI: 1.08–1.97; p=0.013) in TaG1-2 tumors Combination of 3 overexpressed marker is associated with higher tumor grade (p=0.000), a higher tumor number (p=0.04) and is an independent predictor for shorter RFS (HR 3.38, 95% CI: 2.04–5.60; p<0.001) and PFS (HR 8.79, 95% CI: 1.96–39.4; p=0.005) in all NMIBC tumors and shorter RFS in TaG1-2 (HR 3.48, 95% CI: 1.93–6.27; p<0.001) and T1G3 tumors (HR 6.12, 95% CI: 1.70–22.1; p=0.006)

Abbreviations: uPA, urokinase-type plasminogen activator; tPA, tissue-type plasminogen activator; uPAR, uPA receptor; PAI-1, plasminogen activator inhibitor 1; RFS, recurrence free survival; PFS, progression free survival; OS, overall survival; RR, relative risk; HR, hazard ratio; 95% CI, 95% confidence interval.

Table 3 Overview of the Studies Investigating the Plasminogen Activation System (PAS) on Non-Muscle-Invasive Bladder Cancer (NMIBC) and Muscle-Invasive Bladder Cancer (MIBC)

NMIBC and MIBC Tissue
Author	Year	Pat. No.	Study Design	Marker Investigated	Source	Findings
Hasui et al^Citation59	1992	46	not reported	uPA	Tissue	uPA overexpression is associated with higher tumor grade (p<0.01), tumor stage (p<0.01) and a worse CSS (p<0.005)
Seddighzadeh et al^Citation37	2002	194	prospective	uPA, uPAR	Tissue	uPA overexpression is associated with worse CSS (RHR 1.8, 95% CI: 1.0–3.3; p=0.036) and worse OS (RHR 1.9, 95% CI: 1.2–3.0; p=0.006) uPAR overexpression is associated with higher tumor stage (p<0.001), higher risk for metastasis (RHR 4.0, 95% CI: 1.6–9.9; p=0.003), worse CSS (RHR 2.2, 95% CI: 1.3–4.0; p=0.005) and worse OS (RHR 2.2, 95% CI: 1.4–3.5; p=0.001) Combination of overexpressed uPA and uPAR is associated with higher risk for metastasis (RHR 3.2, 95% CI: 1.0–10.2; p=0.049), worse CSS (RHR 2.5, 95% CI: 1.3–4.9; p=0.008) and worse OS (RHR 2.7, 95% CI: 1.6–4.7; p<0.001)
El Kott et al^Citation36	2004	100	retrospective	uPA, uPAR	Tissue	uPA overexpression is associated with higher tumor stage (p=0.00013), lymph node metastasis (p=0.00011) and distant metastasis (p=0.0015) uPAR overexpression is associated with higher tumor stage (p=0.0024), lymph node metastasis (p=0.026), distant metastasis (p=0.0017) and is an independent predictor for worse OS (OR 3.1386; p<0.000001)
Gotanda et al^Citation60	2006	72	retrospective	uPA, PAI-1	Tissue	uPA overexpression is associated with higher tumor stage (p<0.05)
Dohn et al^Citation30	2015	149	retrospective	uPAR	Tissue	uPAR overexpression in cancer cells, macrophages and myofibroblasts is associated with higher tumor grade, higher tumor stage (all p<0.02) and worse OS (HR 2.39, 95% CI: 1.15–5.01; p=0.020) uPAR overexpression in tumor core was associated with female gender (p=0.01)
Dohn et al^Citation35	2015	186	retrospective	uPAR	Tissue	uPAR overexpression in macrophages and myofibroblasts in the tumor core is associated with higher tumor stage and lymph vascular invasion (all p<0.04) uPAR overexpression in macrophages and myofibroblasts at the invasive tumor front is associated with higher tumor stage, higher tumor grade and concomitant CIS (all p<0.03) uPAR overexpression in myofibroblasts at the invasive tumor front is associated with lymph node metastasis (p=0.021)
Chan et al^Citation27	2017	939	retrospective	PAI-1	Tissue	PAI-1 overexpression is associated with higher tumor grade (65% in high grade vs 49% in low grade disease; p<0.001) PAI-1 overexpression independently associated with OS (HR 2.58, 95% CI: 1.52–4.38; p=0.0004) in NMIBC PAI-1 overexpression is independently associated with OS (HR 1.69, 95% CI: 1.09–2.62; p=0.02) in MIBC
Janisch et al^Citation38	2020	272	retrospective	uPA, uPAR, PAI-1	Tissue	uPA overexpression is associated with lymphovascular invasion (p=0.034), lymph node metastasis (p=0.013), shorter RFS (p=0.004), shorter CSS (p=0.012) and is independently associated with RFS (HR 1.75, 95% CI: 1.02–3.00; p=0.041) uPAR overexpression is associated with shorter RFS (p=0.005) and shorter CSS (p=0.007) PAI-1 overexpression is associated with primary muscle invasiveness (p=0.015) and lymphovascular invasion (p=0.024) The number of overexpressed markers is associated with shorter RFS (p<0.001) and worse CSS (p<0.001) while the combination of 2 overexpressed markers is independently associated with RFS (HR 2.25, 95% CI: 1.19–4.27; p=0.013)

Abbreviations: CIS, carcinoma in situ; uPA, urokinase-type plasminogen activator; tPA, tissue-type plasminogen activator; uPAR, uPA receptor; PAI-1, plasminogen activator inhibitor 1; RFS, recurrence free survival; CSS, cancer specific survival; OS, overall survival; HR, hazard ratio; RHR, relative hazard ratio (adjusted HR for specified covariates); OR, odds ratio; 95% CI, 95% confidence interval.

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