Cyclin A Overexpression in Carcinoma of the Renal Pelvis and … · cyclin A to in vito conditions in human TCC to elucidate the potential role of the expression of cyclin A in tumor
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Vol. 3, 1399-1404, August 1997 Clinical Cancer Research 1399
3 The abbreviation used is: TCC, transitional cell carcinoma.
Cyclin A Overexpression in Carcinoma of the Renal Pelvis and
Ureter including Dysplasia: Immunohistochemical Findings in
Relation to Prognosis’
Mutsuo Furihata,2 Yuji Ohtsuki, Hiroshi Sonobe,
Taro Shuin, Akihiro Yamamoto, Naotami Terao,
and Morimasa KuwaharaDepartments of Pathology II [M. F., Y. 0., H. 5.1 and Urology IT. S.].
Kochi Medical School, Nankoku, Kochi 783: Division of Urology,Kochi Takasu Hospital, Kochi 780 [A. Y., N. TI: and Division of
Urology, Fujisaki Hospital, Karatsu, Saga 847 [M. K.l, Japan
ABSTRACTSeveral in vitro studies have shown that cyclin A gene
alteration in the cell cycle plays an important role in carci-nogenesis. We immunohistochemically examined the expres-
sion of cyclin A protein in 120 patients with transitional cell
carcinoma (TCC) of the renal pelvis and ureter, including
adjacent dysplastic lesions to determine their significancefor the tumor behavior and patient prognosis. Cyclin Aimmunostaining of the nucleus was observed in 29 tumors
(24.2%). Furthermore, 17 cyclin A-positive tumors (58.6%)had dysplastic lesions positive for cyclin A antibody. Theprevalence of cases exhibiting cyclin A staining was higherin the high grade (P < 0.01) and invasive tumors (P < 0.05)
than in the other types of tumors. In the selected 117 cases,patients whose TCCs expressed a high level of cyclin Aprotein had a significantly poorer prognosis than those with-
out cyclin A expression (P < 0.01). These in vivo findings
provide the first evidence for frequent and redundant cyclin
A protein overexpression in TCC and suggest that cyclin Aoverexpression is related to the tumor behavior and patient
prognosis. In addition, our observations indicate that over-
expression of cyclin A may be one of the early events, at least
in some cases, in the carcinogenesis of TCC.
INTRODUCTIONDetermination of the sequences of molecular events lead-
ing to and following deregulation of the cell cycle control is
crucial to our understanding of the development and progression
of malignant tumors. Human cyclins, such as A type, B type,
and 01 (C, D, and E) type cyclins, are prime cell cycle-specific
Received 1 1/5/96; revised 4/22/97; accepted 4/24/97.
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.
I This work was supported in part by Grants-in Aid for Scientific
Research from the Ministry of Education, Science, Sports and Culture of
Japan.2 To whom requests for reprints should be addressed. Phone: 0888-66-5811: Fax: 0888-80-2336.
regulators ( 1 ), and they play an important role in the control of
major checkpoints of the cell cycle (2, 3). With the discovery of
inappropriate expression of cyclins in some tumors, it has now
been specifically hypothesized that some cyclins are intimately
involved in oncogenesis, acting as proto-oncogenes (2, 3). Di-
verse patterns of redundant expression of particular cyclins,
such as cyclins Dl and E, in different tumor cell lines have
previously been reported in oncogenesis (4-14). A relationship
of cyclin A abnormalities to carcinogenesis has also been no-
ticed. Human cyclin A gene at a site of hepatitis B virus DNA
integration was identified in a hepatocellular carcinoma in as-
sociation with abnormality of the ct-c/in A gene ( I 5, 16). Cyclin
A also associates with adenovirus E1A oncoprotein ( I 7, 18) and
has been identified in complexes containing the E2F transcrip-
tion factor and p107, which is structurally related to the retino-
blastoma protein (1 8 -22). Although little is known concerning
the significant role of cyclin A in tumorigenesis, these studies
suggest that cyclin A may be of value in identifying proliferative
tumor cells, and its alteration may be an important step in the
pathogenesis of certain cell lines and primary tumors.
There is little data available concerning whether premalig-
nant states, including dysplasia, in the transitional epithelium
progress to TCC.3 However, most reports have indicated that
dysplasia acts as a common precursor to TCC because it shows
a high propensity to progress to carcinoma in situ (23, 24). In
addition, the presence of dysplasia adjacent to TCC is associated
with an increased risk of subsequent recurrence and progression.
(25). Therefore, the features of dysplasia provide ideal systems
to study the proposed multiple-step nature of carcinogenesis.
Molecular and histological studies of preinvasive urothelial be-
sions have frequently demonstrated genomic and pathological
alterations in carcinoma in situ and dysplastic lesions, suggest-
ing that at least some of these abnormalities occur very early in
oncogenesis (26, 27). Recently, increased expression of cyclin
Dl protein was reported in precancerous lesions of coborectal
mucosa (28). These findings prompted us to study the level of
expression of cyclin A protein at different stages along the
multistage process of urothelial carcinogenesis from normal
mucosa to dysplasia and carcinoma.
In the present study, we extended these observations of
cyclin A to in vito conditions in human TCC to elucidate the
potential role of the expression of cyclin A in tumor develop-
ment and to assess their prognostic value, also examining the
dysplastic epithelium adjacent to the tumor. The relationship
with various clinicopathobogical factors was then analyzed.
El � Patianta with non- invasiva tvna nf TC(� (n =
1 2 3 4 5 6 iYEARS AFTER NEPHROURETERECTOMV
9 10
> a cyckn A-negative (n -68)
YEARS AFTER NEPHROURETERECTOMV
10
1402 Cyclin A in Transitional Cell Carcinoma
100
90
80
70
�60
� so
�4o
20
10
0
20
10
A-positiv.(n-
B : Patients with low arade TCC (n = 721
i 2 3 4 5 6 7 8 9 i�o
YEA� AFTER NEPHROURETERECTOMy
100 1 �
90
80
70
�6o�5o
20
10
Fig. 3 The cumulative Kaplan-Meier survival curves of patients with carcinoma of renal pelvis and ureter. There are statistically significant
differences between the cyclin A-positive and cyclin A-negative group in all 117 patients tested (A, P < 0.01) and in patients with low grade TCC(B, P < 0.05) and high grade TCC (C, P < 0.05). Although no significant difference in the survival rate for the patients with noninvasive type ofTCC is detected (D), there is a statistically significant difference between the cyclin A-positive and cyclin A-negative group for the patients with the
invasive type of ICC (E. P < 0.05).
erative 9-year survival rate of the cyclin A-positive group (n =
17) was 13.8%, whereas that of the cyclin A-negative group
(n 28) was 48.2%; the difference between these rates was also
statistically significant (P < 0.05; Fig. 3C). In the tumor growth
pattern, there was no significant difference in the survival rate
for the 25 patients with the noninvasive type of TCC between
cyclin A-positive (n = 4) and cyclin A-negative groups (n = 21;
Fig. 3D). In contrast, the postoperative 10-year survival rates for
the 92 patients with the invasive type of TCC was 18.6% for the
cyclin A-positive group (n = 24) and 50.3% for the cyclin
A-negative group (n 68), respectively. There was a significant
difference between these groups (P < 0.05; Fig. 3E).
suggest that cyclin A overexpression may be associated with the
clinical behavior of high grade and progressive TCCs. In the
present study, our findings also indicated that the groups with a
high level of cyclin A expression was associated with a signif-
icantly poorer prognosis than the groups without its expression.
These in vivo findings suggest that cyclin A overexpression is
also related to the poor prognosis for the patients with renal
pelvic and ureteral TCCs.
We also investigated the expression of cyclin A in
samples containing normal and dysplastic lesions adjacent to
the primary tumors. One important finding in this study was
that approximately 58% (17 of 29) of the dysplastic lesions
adjacent to cyclin A-positive tumors eventually featured cy-
din A overexpression. It is also of interest to note that
physiological levels of cyclin A protein was detected in the
subsets of cells of the basal layer in the nonneoplastic tnan-
sitional epithelium. This observation supports the importance
of abnormal levels of expression of this protein in urothebiab
dysplasia. In the present study, these findings suggest that
unscheduled overexpression ofcycbin A may be due to loss of
the cell cycle regulation at an early stage of the pathogenesis
of some cases of urinary neoplasms. In addition, detection of
cyclin A overexpression in premalignant lesions of the un-
nary tract may potentially be useful in diagnosis and deter-
mination of their roles in the natural history of ICC.
Although the molecular basis for the positive immuno-
staining is still indefinite, this is the first report of frequent
cyclin A protein overexpression in a large series of primary
human TCCs including dysplasia. In addition, the present study
also demonstrated that immunohistochemical detection of cyclin
A protein is useful in the search for novel and potentially useful
prognostic markers in ICC also. Recent reports have further
demonstrated the presence of a link between oncogenesis and
the cell cycle by correlating the deranged expression of cyclins
to the loss of growth control (4, 6, 14). Theretbre, the frequent
alterations of cyclin A protein in tumor cells are also suggestive
of its role as an oncogene. Up to the present time, most inves-
tigations into the relationship between the expression of cyclin
and carcinogenesis were mainly carried out in vitro using certain
cell lines. Further comprehensive studies using excised human
carcinoma tissue samples from greater numbers of human tu-
mors and including measurement of DNA and/or RNA levels
will be needed to elucidate the function and clinical significance
of cyclin A overexpression.
ACKNOWLEDGMENTS
We are grateful to Dr. T. Moniki for providing clinical materials.
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