Korean J Urol.  2006 Apr;47(4):426-435. 10.4111/kju.2006.47.4.426.

Effects of Transforming Growth Factor-beta1 and Its Receptor on the Development, Recurrence and Progression of Human Bladder Cancer

• Quan C
• Park MS
• Jo SW
• Lee SC
• Kim WJ

• KMID: 1997155
• DOI: http://doi.org/10.4111/kju.2006.47.4.426

Abstract

PURPOSE
We investigated whether the expression levels of Transforming growth factor beta1 (TGF-beta1) and its receptors were related to the development, recurrence, progression and disease-free survival in the patients with bladder cancer.
MATERIALS AND METHODS
The mRNA levels of TGF-beta1 and its receptors were examined in 102 tumor specimens from patients with primary bladder cancer, 29 corresponding normal bladder mucosae specimens surrounding these tumors and 15 normal bladder mucosae specimens by performing quantitative competitive PCR (QC-PCR). The protein levels of TGF-beta1 and its receptors were investigated by performing immunohistochemical staining on sections cut from 86 archival bladder tissue paraffin blocks.
RESULTS
QC-PCR analysis showed that expressions of TGF-beta1, TGF-beta receptor I (TGF-betaRI) and receptor II (TGF-betaRII) in the superficial and low-grade bladder cancers were significantly higher than those in both the corresponding normal bladder mucosae surrounding the cancer (p= 0.0069, 0.0022 and 0.0046, respectively) and the control's normal bladder mucosae (p=0.0014, 0.0125 and 0.0089, respectively). Expressions of TGF-beta1 and its receptors were enhanced in the non-recurred and non-progressed patients compared to the recurred cases (p=0.0022, 0.0003 and 0.0001, respectively) and the progressed cases (p=0.0002, <0.0001 and <0.0001, respectively). Patients with high expression of TGF-beta and its receptors had a significantly higher disease-free survival rate than those patients with low expressions (p=0.0129, 0.0121 and 0.0132, respectively).
CONCLUSIONS
The enhanced expression of TGF-beta1 and its receptors was correlated not only with superficial and low-grade bladder cancer, but also with enhanced patient survival. In conclusion, our findings suggest that the expressions of TGF-beta1 and its receptors are useful prognostic markers for a patient's resistance to disease recurrence and/or progression.

Keyword

Transforming growth factor-beta1; TGF beta Receptor; Bladder cancer

MeSH Terms

Disease-Free Survival
Humans*
Mucous Membrane
Paraffin
Polymerase Chain Reaction
Receptors, Transforming Growth Factor beta
Recurrence*
RNA, Messenger
Transforming Growth Factor beta
Transforming Growth Factor beta1
Urinary Bladder Neoplasms*
Urinary Bladder*
Paraffin
RNA, Messenger
Receptors, Transforming Growth Factor beta
Transforming Growth Factor beta
Transforming Growth Factor beta1

Figure

• 

  Fig. 1 A-C, Each serially diluted competitor (1µl) is co-amplified with a 1µl sample of cDNA. The PCR products are separated on 2% agarose gel and then visualized by ultraviolet light after ethidium bromide staining, and the density of each band is quantified. D-F, The ratio of target cDNA to its competitor is log transformed and plotted against the amount of competitor cDNA initially added to each PCR reaction to produce the standard curves. When the competitor is diluted to 10-6 (50pg/ml), the ratio of TGF-β1 to competitor approaches 1. When the competitor is diluted to 5×10-7 for TGF-βRI (3pg/ml) and TGF-βRII (25pg/ml), the ratio of the target to competitor reaches 1. Therefore, we perform QC-PCR using the competitor at the above concentrations for quantifying TGF-β1 and its receptor expression levels (PCR: polymerase chain reaction, TGF-β1: transforming growth factor β1, TGF-βR: transforming growth factor β receptor).

• 

  Fig. 2 Positive correlation among the mRNA expression levels of TGF-β1, TGF-βRI and RII in bladder cancer. A: TGF-β1 vs TGF-βRII (r=0.8780, p<0.0001), TGF-β1 vs TGF-βRI (r=0.7468, p<0.0001), B: TGF-βRI vs TGF-βRII (r=0.7672, p<0.0001).

• 

  Fig. 3 The expression levels of TGF-β1 A, TGF-βRI B and RII C mRNA are strongly correlated with the length of disease free survival of the bladder cancer patients.

• 

  Fig. 4 Immunostaining of TGF-β1, TGF-βRI and TGF-βRII in normal and cancerous bladder tissues. Each column shows sections from the same tissue sample that were stained for the different antigens. The left column shows similar levels of the three proteins in normal bladder mucosa (A: TGF-β1, B: TGF-βRI, C: TGF-βRII). The middle column shows elevated levels of the three proteins in grade 1/superficial bladder cancer (D: TGF-β1, E: TGF-βRI, F: TGF-βRII). The right column shows similar low levels of the three proteins in grade 3/invasive bladder cancer (G: TGF-β1, H: TGF-βRI, I: TGF-βRII).

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