Recent clinical trials have demonstrated how, despite radical surgery and adjuvant chemotherapy, more than half of muscle invasive bladder cancers develops hematogenous spread and distant metastases. Furthermore, a high percentage of patients with superficial disease has progression to higher stage tumors, which is difficult to predict on the basis of clinical features (1).

In a recent report by Gazzaniga et al.(2), we investigated the utility of an EGFR1-based RT-PCR assay to detect circulating cancer cells in the peripheral blood collected before surgery from a subset of bladder cancer patients. The finding of EGFR mRNA in blood drawings performed in nonmetastatic patients who relapsed early lead us to the hypothesis of the utility of such an assay in the pre-surgical staging of bladder cancer patients.

In the period after the publication of the article, we sought to investigate whether EGFR mRNA expression was modified in blood samples collected from the same patients 3 months after surgery and from patients with locally advanced tumors 1 year after chemotherapy. RNA extraction from whole blood and RT-PCR with EGFR-specific primer sequences were performed as described previously (2).

The results of EGFR amplification in blood samples collected after surgery and after adjuvant therapy were then compared with that obtained in the same patients before surgical treatment. The comparative analysis between EGFR amplifications in blood samples collected before and after surgical treatment revealed that EGFR mRNA expression was invariable in all patients except 2, who were EGFR negative before transurethral resection and resulted EGFR positive 3 months later. Follow-up data revealed in both these patients a progression of disease, as shown in Table 1.

In most patients with higher staged tumors, who had shown high EGFR expression levels in blood before surgery, we observed the absence of EGFR-specific amplifications in blood drawings performed 1 year after adjuvant cisplatinum-based chemotherapy. All these patients are at present in stability of disease.

In 2 of these metastatic patients, however, EGFR mRNA expression in blood samples collected before and after therapy remained invariable; both these patients died because of the disease (Fig. 1).

These results may represent an additional confirmation to the utility of EGFR-based RT-PCR assay in monitoring patients with bladder cancer also in a postsurgical phase, as well as in the clinical management of patients included in adjuvant therapy protocols. The detection of EGFR expressing circulating cells may allow the selection of patients at high risk to relapse, thus enhancing the chance of cure. Furthermore, the use of EGFR-based RT-PCR assay in patients in course of adjuvant treatment may allow the detection of early recurrence and treatment failure, thus allowing new approaches with salvage therapies.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

      
1

The abbreviations used are: EGFR, epidermal growth factor receptor; RT-PCR, reverse transcription-PCR.

Fig. 1.

Expression of EGFR in blood samples from patients with bladder cancer. A, EGF-RT-PCR in blood samples collected before surgery; B, EGF-RT-PCR in blood samples collected after surgery; C, EGFR RT-PCR in blood samples collected after chemotherapy. Lanes 1 and 4, patients with superficial disease; Lanes 8, 10, 12, 13, and 16, patients with locally advanced disease (see Table 1).

Fig. 1.

Expression of EGFR in blood samples from patients with bladder cancer. A, EGF-RT-PCR in blood samples collected before surgery; B, EGF-RT-PCR in blood samples collected after surgery; C, EGFR RT-PCR in blood samples collected after chemotherapy. Lanes 1 and 4, patients with superficial disease; Lanes 8, 10, 12, 13, and 16, patients with locally advanced disease (see Table 1).

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Table 1

EGFR mRNA expression before surgery, after surgery, and after chemotherapy in patients with bladder cancer

Patient no.StageEGFR mRNA expressionFollow-upa
TumorPeripheral blood
Pre-surgical treatmentbPostsurgical treatmentcPost-chemotherapyd
0is − − − Free 
0a − − − Free 
0a − − − Free 
0a − − Progr. st. I 
− − Progr. st. II 
Free 
II − Stability 
II − − − − Stability 
III − Stability 
10 III − Stability 
11 III − Stability 
12 III Death 
13 III − Stability 
14 III − Stability 
15 III − Stability 
16 III Death 
Patient no.StageEGFR mRNA expressionFollow-upa
TumorPeripheral blood
Pre-surgical treatmentbPostsurgical treatmentcPost-chemotherapyd
0is − − − Free 
0a − − − Free 
0a − − − Free 
0a − − Progr. st. I 
− − Progr. st. II 
Free 
II − Stability 
II − − − − Stability 
III − Stability 
10 III − Stability 
11 III − Stability 
12 III Death 
13 III − Stability 
14 III − Stability 
15 III − Stability 
16 III Death 
a

Thirty months after surgery.

b

Days (1–2) before surgery.

c

Three months after surgery.

d

One year after chemotherapy.

1
Ozen H., Graig Hall M. Bladder cancer.
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2
Gazzaniga P., Gandini O., Giuliani L., Magnanti M., Gradilone A., Silvestri I., Gianni W., Gallucci M., Frati L., Aglianò A. M. Detection of epidermal growth factor receptor mRNA in peripheral blood: a new marker of circulating neoplastic cells in bladder cancer patients.
Clin. Cancer Res.
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2001
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