close

Вход

Забыли?

вход по аккаунту

?

786

код для вставкиСкачать
550
Differential Retinoblastoma and p16INK4A Protein
Expression in Neuroendocrine Tumors of the Lung
Hirotoshi Dosaka-Akita, M.D., Ph.D.1
Philip T. Cagle, M.D.2
Hiromitsu Hiroumi, M.D.1
Masahiro Fujita, M.D., Ph.D.3
Motoyuki Yamashita, M.D., Ph.D.4
Anupama Sharma, M.D.5
Yoshikazu Kawakami, M.D., Ph.D.1
William F. Benedict, M.D.4
1
First Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
2
Department of Pathology, Baylor College of Medicine, Houston, Texas.
3
Department of Pathology, National Sapporo Hospital, Sapporo, Japan.
4
Department of Genitourinary Medical Oncology,
University of Texas M. D. Anderson Cancer Center,
Houston, Texas.
5
Department of Pathology, Duke University Medical Center, Durham, North Carolina.
Supported in part by National Institute of Health
Grant CA-54672 to William F. Benedict and by a
Grant-in-Aid (No. 11670558) from the Minister of
Education, Science and Culture of Japan to Hirotoshi Dosaka-Akita.
The authors thank Michio Shimizu, Department of
Pathology, Hokkaido University Medical Hospital;
Kenzo Okamoto, Department of Pathology, Hokkaido Kin-ikyou Chuo Hospital; Takehito Nakabayashi, Department of Respirology, National Sapporo Hospital; Tamotsu Hirata, Department of
Surgery, National Sapporo Minami Hospital; Yoshikazu Araya, Department of Respirology, National Hakodate Hospital; Tetsuo Shimizu and
Toshiaki Fujikane, Department of Medicine, National Dohoku Hospital; Masato Hashimoto, Department of Surgery, Iwamizawa Rousai Hospital;
Takashi Yoshikawa, First Department of Medicine,
Obihiro Kousei Hospital; and Kazuo Takaoka, Department of Respirology, Nikkou Memorial Hospital; for resected tumors and clinical data.
Address for reprints: Hirotoshi Dosaka-Akita, M.D.,
Ph.D., First Department of Medicine, Hokkaido University School of Medicine, North 15, West 7,
Kita-ku, Sapporo, 060-8638, Japan.
Received June 18, 1999; revision received October
6, 1999; accepted October 6, 1999.
© 2000 American Cancer Society
BACKGROUND. Neuroendocrine neoplasms of the lung represent a wide spectrum
of phenotypically and biologically distinct entities. Their histopathologic diagnosis,
which carries therapeutic and prognostic significance, may sometimes be difficult
because of their overlapping features. We previously demonstrated that large cell
neuroendocrine carcinomas (LCNECs) and small cell lung carcinomas (SCLCs)
failed to show positive nuclear staining of RB protein (RB⫺), whereas typical and
atypical carcinoids (TCs and ACs) showed nuclear RB immunostaining (RB⫹).
METHODS. In the current study, a series of 58 surgically resected lung tumors, of
which 33 tumors were initially diagnosed as SCLCs and 25 as TCs or ACs, were
studied for RB and p16 protein expression by immunohistochemistry. They were
also reviewed for their pathologic diagnosis; the reviewers were blinded to the RB
and p16 protein status.
RESULTS. Nineteen tumors were diagnosed as TCs, 5 as ACs, 7 as LCNECs, and 27
as SCLCs. Three of seven LCNECs were RB⫹, whereas the other four were RB⫺. In
contrast, all 19 TCs were RB⫹ and all 27 SCLCs were RB⫺. In addition, two of five
ACs were RB⫹, whereas the other three were RB⫺. Interestingly, all 3 RB⫹ LCNECs
and the 1 RB⫹ AC tested failed to show nuclear staining of p16 protein in any
tumor cells (p16⫺), although some normal stromal cells showed nuclear staining of
p16 protein (p16⫹) as positive internal controls, indicating loss of p16 function in
these tumors. It is also noteworthy that the three RB⫹ LCNECs were initially
diagnosed as SCLCs and one of the RB⫺ ACs was initially considered a TC. With the
exception of TCs, tumors were significantly more prevalent among heavy smokers
with ⬎20 pack-years compared with nonsmokers and light smokers with ⱕ20
pack-years (P ⬍ 0.01).
CONCLUSIONS. These findings suggest that all SCLCs and LCNECs have abnormalities in the p16:RB pathway, as do at least certain ACs, whereas the p16:RB pathway
is normal in TCs. Cancer 2000;88:550 –56. © 2000 American Cancer Society.
KEYWORDS: neuroendocrine tumors of the lung, retinoblastoma protein, p16INK4A
protein, diagnostic marker, immunohistochemistry.
N
euroendocrine (NE) neoplasms of the lung represent a wide spectrum of pathologically and biologically distinct entities.1– 4 Of
these, atypical carcinoids (ACs) and large cell neuroendocrine carcinomas (LCNECs) are much less commonly encountered neoplasms
than the other histologic types of NE tumors, including typical carcinoids (TCs) and small cell lung carcinomas (SCLCs). In the past,
LCNECs may have been classified as either atypical carcinoids or
SCLCs because of their overlapping features.5,6 It has been recently
indicated that 1) classification of NE tumors of the lung is most
reproducible for classification of TC and SCLC but less reproducible
for AC and LCNEC, and 2) a need for more careful definition and
application of criteria for TC versus AC and for SCLC versus LCNEC.7
Moreover, the correct histopathologic diagnosis of specific NE tumors
may be important for the consideration of different therapeutic interventions and may have potential prognostic implications.6,8
RB and 16NK4A Proteins in Neuroendocrine Lung Tumors/Dosaka-Akita et al.
It has been reported that the frequency of tumor
suppressor gene alterations, including those of the p53
and retinoblastoma (RB) genes, varies among categories of NE tumors.9 –22 Provided that the identification
of a specific alteration is consistently and reproducibly
distinctive among specific NE tumors, such alterations
might be useful for the differential diagnosis of NE
tumors of the lung.
The RB gene is a prototypical tumor suppressor
gene encoding a nuclear phosphoprotein with a molecular weight of 105–110 kD.23 The functional loss of
RB protein is believed to be a key event in the development of a variety of neoplasms, including SCLCs.24
Most RB gene alterations result in the loss of RB protein expression or in a truncated RB protein, which
does not enter the nucleus. Heterogeneous positive
nuclear RB immunostaining is, in general, indicative
of normal RB function, whereas negative intranuclear
RB immunostaining in all tumor cells reflects functional loss of the RB gene.25,26 Moreover, to date, the
presence or absence of normal RB protein expression
determined by immunohistochemistry has been
found to be the most sensitive and specific method for
determining RB status in a given tumor.27 We have
previously demonstrated that SCLCs and LCNECs
failed to show positive nuclear staining of RB protein
by immunohistochemistry; in contrast, TCs and ACs
showed nuclear RB immunostaining, although the latter had a stronger and more homogeneous staining
pattern than the former.28
p16INK4A (p16) protein, the product of the CDKN2
gene, which has been found to be deleted in a variety
of tumor cells,29,30 inhibits CDK4- and CDK6-mediated phosphorylation of RB protein.31,32 Therefore,
p16 and RB proteins are suggested to function in a
single regulatory pathway of the cell cycle and tumor
suppression,31,32 which is supported by observation of
an inverse correlation between alterations of both
proteins in primary lung carcinomas and lung carcinoma cell lines.33–37 Immunohistochemical detection
of p16 protein is a sensitive and specific method of
screening for p16 alterations resulting from both homozygous deletion and DNA hypermethylation.37 We
have recently reported that RB-negative bladder tumors exhibited strong nuclear p16 staining, whereas
each tumor showing strong, homogeneous nuclear RB
staining (overexpression) was p16 negative.38 These
findings support the idea that loss of p16 protein function could be related to RB overexpression, since p16
can induce transcriptional down-regulation of RB and
its loss may lead to aberrant regulation of RB expression. Conversely, loss of RB function has been associated with high p16 protein expression in several types
of tumors, including nonsmall cell lung carcinomas.37
551
In the current study, we investigated expression of
RB and p16 proteins in surgically resected NE tumors
of the lung, and showed the potential implication of
both proteins in their diagnosis.
MATERIALS AND METHODS
Tumor Specimens
A total of 58 lung tumor specimens, which had been
surgically resected and initially diagnosed at surgery
as carcinoids (25 tumors) or SCLCs (33 tumors) between 1979 and 1996, were collected from a cohort of
surgically resected lung tumor specimens and analyzed in the current study. For all 33 tumors diagnosed
as SCLC, biopsies and cytologic examination before
surgery were not diagnostic, and the diagnosis of
SCLC was made after surgery only by subsequent
pathologic analysis. The tumors were obtained at
Hokkaido University Medical Hospital, National Sapporo Hospital, National Sapporo Minami Hospital,
National Hakodate Hospital, National Dohoku Hospital, Iwamizawa Rosai Hospital, Obihiro Kosei Hospital,
Hokkaido Kin-ikyo Chuo Hospital, and Nikko Memorial Hospital, all located in Hokkaido, Japan, and evaluated by different pathologists at the time of surgery
as routine pathologic practice in each hospital. In this
cohort, 19 tumors were diagnosed as TCs, 6 as ACs,
and 33 as SCLCs. Subsequently, the 58 tumors were
analyzed for their RB and p16 protein expression and
were also independently reclassified by one of us
(P.T.C.) according to established histopathologic criteria28 and blinded to the RB and p16 protein status.
Briefly, the tumors were evaluated histologically for a
spectrum of features, including organoid pattern, mitoses, necrosis, nuclear pleomorphism, nucleoli, nuclear-to-cytoplasmic ratio, nuclear chromatin, and evidence of neuroendocrine differentiation. Clinical and
clinicopathologic data at surgery were available on all
58 tumors.
Immunohistochemical Analysis
Immunohistochemical staining for RB and p16 proteins was performed on formalin fixed, paraffin embedded tissue sections. The methods for staining RB
nuclear protein have been described previously.26,39
Briefly, after deparaffinization and hydrogen peroxide
treatment of 5 ␮m sections from formalin fixed, paraffin embedded tissue blocks, they were processed for
immunostaining for RB protein with an affinity-purified polyclonal anti-RB antibody, RB-WL-1,40 at a dilution of 1:628. The RB immunostaining pattern for
each case was independently evaluated by two investigators (H.D.-A., W.F.B.). Tumors were scored for the
presence or absence of nuclear RB staining using criteria previously established.41 The entire specimens
552
CANCER February 1, 2000 / Volume 88 / Number 3
were examined at ⫻100 and ⫻400 magnifications. To
be considered adequate, a tumor section had to contain numerous normal RB positive stromal cells as
internal controls. To be scored RB negative (RB⫺), all
tumor cells in the section had to show no nuclear RB
staining when there were adjacent RB positive normal
stromal cells in the same tumor section. Tumors were
scored as RB positive (RB⫹) if several tumor nuclei
had RB staining, although in all RB⫹ cases the majority of tumor cells had some nuclear staining in this
cohort. Results of the RB immunostaining patterns
were then compared with the histopathologic diagnosis category and with the clinical and clinicopathologic data.
Immunohistochemical staining of p16 nuclear
protein was performed as previously described.38
Briefly, after deparaffinization, hydrogen peroxide
treatment, and antigen retrieval procedure of 5 ␮m
sections from formalin fixed, paraffin embedded tissue blocks, they were processed for immunostaining
for p16 protein with a mouse monoclonal antibody
against human p16 protein, NCL-p16, clone DCS-50
(Vector Laboratories, Burlingame, CA) at a 1:25 dilution. The p16 immunostaining pattern for each case
was independently evaluated by two investigators
(H.D.-A., W.F.B.). The criteria for the evaluation of the
presence or absence of nuclear p16 staining (p16⫹ or
p16⫺) were the same as those described previously for
RB protein.
Statistical Analysis
The associations between histologic types and various
characteristics were analyzed by the chi-square test or
the Fisher exact test as appropriate.42 The association
between histologic types and age was analyzed by the
Student t test. The significance level chosen was P ⬍
0.05, and all tests were two-sided.
RESULTS
A series of surgically resected 58 lung tumors were
reevaluated for their pathologic diagnosis. Nineteen
tumors initially diagnosed as TCs at surgery were reevaluated and determined to be 18 TCs and 1 AC; 6
tumors initially diagnosed as ACs were reevaluated
and found to be 1 TC, 4 ACs, and 1 LCNEC; and 33
tumors initially diagnosed as SCLCs were reevaluated
and found to be 6 LCNECs and 27 SCLCs. Therefore,
19 tumors were subsequently diagnosed as TCs, 5 as
ACs, 7 as LCNECs, and 27 as SCLCs by pathologic
reevaluation in this study (Table 1).
These tumors were studied for their RB and p16
protein expression by immunohistochemistry. Three
of seven LCNECs were RB⫹ (Fig. 1C) and the other
four were RB⫺, whereas numerous RB⫹ normal stro-
TABLE 1
Pathologic Diagnosis and RB/p16 Protein Status in Neuroendocrine
Tumors of the Lung
Diagnosis at surgery
TC 19
AC 6
SCLC 33
Diagnosis in
this study
TC 18
ACa 1
TCa 1
AC 4
LCNECa 1
LCNECa 6
SCLC 27
RB/p16 status
⫹/⫹ or NE 18
⫺/⫹b 1
⫹/⫹ 1
⫺/⫹ or NE 2
⫹/⫺ or NE 2
⫺/⫹ 1
⫹/⫺b 3
⫺/⫹ 3
⫺/⫹ 27
TC: typical carcinoid; AC: atypical carcinoid; SCLC: small cell lung carcinoma; NE: not evaluable
because of the loss of adequate p16 protein expression in positive internal control cells (normal stromal
cells).
a
Diagnosis reclassified by P.T.C.
b
Initial diagnosis at surgery was inconsistent with RB/p16 status.
mal cells were present in every section as internal
controls (Fig. 1A). Two of five ACs were RB⫹ and the
other three, one of which contained several cells with
a polygonal shape and more vesicular nuclei, reminiscent of LCNEC, were RB⫺, although again numerous
RB⫹ stromal cells were present in the sections. In
contrast, all 19 TCs were RB⫹ and all 27 SCLCs were
RB⫺. This difference in the loss of RB protein among
specific NE tumors was statistically significant (TCs vs.
ACs, P ⬍ 0.01; TCs vs. LCNECs, P ⬍ 0.01; TCs vs.
SCLCs, P ⬍ 0.01; ACs vs. SCLCs, P ⬍ 0.01; LCNECs vs.
SCLCs, P ⬍ 0.01) (Table 2). No differences in the
histologic features were found between RB⫹ and RB⫺
LCNECs and between RB⫹ and RB⫺ ACs.
The reciprocal loss of p16 and RB proteins has
been reported in primary lung carcinomas and lung
carcinoma cell lines,33–37 to support the hypothesis
that both proteins function in a single regulatory pathway of the cell cycle and tumor suppression.31,32 Interestingly, all three RB⫹ LCNECs and the one RB⫹
AC tested failed to show nuclear staining of p16 protein in any tumor cells, although some normal stromal
cells were p16⫹ as positive internal controls (Fig. 1D;
Table 3), indicating loss of p16 function in these tumors. All four RB⫺ LCNECs and the two RB⫺ ACs
examined manifested strong p16 immunostaining
(Fig. 1B; Table 3), which was also observed in SCLCs.
This difference in the loss of p16 protein among NE
tumors was statistically significant (ACs vs. SCLCs, P ⬍
0.01; LCNECs vs. SCLCs, P ⬍ 0.01) (Table 2). These
findings suggest that all SCLCs and LCNECs have abnormalities in the p16:RB pathway, as do at least certain ACs, by loss of either RB or p16 protein in LCNECs
and ACs and by loss of RB protein in SCLCs (Tables 2
RB and 16NK4A Proteins in Neuroendocrine Lung Tumors/Dosaka-Akita et al.
553
FIGURE 1. Immunohistochemical staining of RB (A and C) and p16 proteins (B and D) in large cell neuroendocrine carcinomas is shown. The staining pattern
of an RB⫺/p16⫹ tumor is shown in A and B, and staining for an RB⫹/p16⫺ tumor is shown in C and D. A representative RB⫹ normal stromal cell is shown in
A (large arrow) and two p16⫹ stromal cells are seen in D (small arrows) as internal controls.
and 3). Of note, one RB⫹/p16⫹ tumor initially diagnosed as AC at surgery was found to be a TC. Moreover, one RB⫺/p16⫹ tumor initially diagnosed as TC
was found to be an AC, and three RB⫹/p16⫺ tumors
initially diagnosed as SCLCs were found to be LCNECs
by pathologic reevaluation blinded to the RB and p16
protein status (italic in Table 1).
The associations between histologic types of NE
tumors and clinical and clinicopathologic characteristics were next analyzed (Table 2). Patients with LCNECs and SCLCs were significantly older than those
with TCs (P ⫽ 0.03 and P ⬍ 0.01, respectively),
whereas patients with ACs were older but not significantly older than those with TCs. All three patients
with ACs, 6 of 7 with LCNECs, and 22 of 26 with SCLCs
whose smoking habits could be identified were heavy
smokers with more than 20 pack-years, whereas only 2
of 13 patients with TCs were heavy smokers (P ⬍ 0.01).
DISCUSSION
In the current study, we showed that all SCLCs and
LCNECs had abnormalities in the p16:RB pathway, as
did at least certain ACs, by loss of RB protein in SCLCs
and by reciprocal loss of either RB or p16 protein in
LCNECs and ACs, whereas the p16:RB pathway was
normal in TCs. In the previous study,28 using resected
specimens, wedge biopsies, and bronchoscopic biopsies, we showed the distinct differences in RB expression in SCLCs and LCNECs compared with TCs and
ACs: 40 SCLCs and 6 LCNECs failed to show RB staining, whereas 44 TCs and 15 ACs manifested RB staining, although the latter showed a stronger and more
homogeneous RB staining pattern than the former.
Based on the recent findings in bladder carcinomas in
which strong and homogeneous staining of RB protein
was associated with loss of p16 protein function,38
many of the previously reported ACs may have been
554
CANCER February 1, 2000 / Volume 88 / Number 3
TABLE 2
Clinical and Clinicopathologic Characteristics of RB-Stained and p16-Stained Neuroendocrine Tumors of the
Lung
Characteristics
TCs
ACs
LCNECs
SCLCs
P valuea
Age (yrs, mean ⫾ SD)
(Range)
Gender
Male
Female
Smoking (pack-years)b
0–20
⬎20
pStagec
I
II
IIIa
IIIb
IV
RB
(⫹)
(⫺)
p16d
(⫹)
(⫺)
50.2 ⫾ 14.0
21–67
60.4 ⫾ 12.1
45–72
63.7 ⫾ 10.9
46–82
62.1 ⫾ 8.9
36–74
⬍0.01 (TCs/SCLCs)
0.03 (TCs/LCNECs)
11
8
5
0
6
1
21
6
NS
11
2
0
3
1
6
4
22
⬍0.01
12
1
1
0
0
3
0
0
0
1
2
2
1
0
1
9
7
8
1
2
NS
19
0
2
3
3
4
0
27
⬍0.01
7
0
2
1
4
3
27
0
⬍0.01
a
P values determined by the Student t test for age, and P values by the chi-square test or the Fisher exact test as appropriate for others. NS: not statistically
significant.
b
Data on smoking habits were available on patients with 13 TCs, 3 ACs, 7 LCNECs, and 26 SCLCs.
c
Data on pStage were available in 14 TCs, 4 ACs, 6 LCNECs, and 27 SCLCs.
d
Not evaluable for 12 TCs and 2 ACs because of the loss of adequate p16 protein expression in positive internal control cells (normal stromal cells).
TABLE 3
RB and p16 Protein Expression in ACs and LCNECs
Tumors
ACs
# 13-4
# 13-6
# 17-2
# 18-3
# 18-4
LCNECs
# 2-3
# 3-5
# 7-1
# 13-5
# 1-2
# 3-3
# 3-4
RB
p16
⫺
⫺
⫺
⫹
⫹
⫹
NEa
⫹
NE
⫺
⫺
⫺
⫺
⫺
⫹
⫹
⫹
⫹
⫹
⫹
⫹
⫺
⫺
⫺
RB: retinoblastoma; ACs: atypical carcinoids; LCNECs: large cell neuroendocrine carcinomas.
a
Not evaluable because of the loss of adequate p16 protein expression in positive internal control cells
(normal stromal cells).
p16⫺ in retrospect and need to be evaluated for their
p16 status. The current study indicates that LCNECs
could have intact RB protein, but all such RB⫹
LCNECs had lost p16 function. In addition, AC may
show reciprocal loss of RB and p16 proteins as men-
tioned above, whereas all of the SCLCs studied by us
to date have lost RB function as reported in the previous28 and current studies.
Accurate diagnosis of specific NE tumors may influence the therapy given and the clinical outcome.6,8
In this sense, different patterns of RB and p16 protein
expression between TCs and ACs and between LCNECs and SCLCs determined by immunohistochemistry may be helpful in the differential diagnosis of
specific NE tumors. It is sometimes difficult to distinguish AC from TC and LCNEC from SCLC even in
surgically resected specimens,7 and these tumors are
often diagnosed based on bronchoscopic biopsies,
fine-needle aspiration biopsies, or biopsies of lymph
node metastases, which are smaller and more crushed
compared with surgically resected specimens. The
separation of AC from TC is based on numbers of
mitotic figures and necrosis as well as disorganized
architecture, cellularity, and cytologic atypia.43 It may
remain difficult, therefore, in certain cases, to distinguish AC from TC in a given tumor, even though new
criteria for separation of AC from TC has been recently
proposed.8 LCNECs generally have more cytoplasm
than SCLCs and are unlikely to be spindle cells but
show considerable overlap in other features with
RB and 16NK4A Proteins in Neuroendocrine Lung Tumors/Dosaka-Akita et al.
SCLCs, including organoid pattern, a high frequency
of mitosis, and areas of necrosis. Consequently, the RB
and p16 protein status determined by immunohistochemistry may on occasion be a diagnostic marker for
ACs versus TCs and for LCNECs versus SCLCs. Discrepant RB and p16 immunostaining patterns for TCs,
such as either negative RB or p16 immunostaining,
and for SCLCs, such as positive RB and negative p16
immunostaining, may be a basis for reevaluation of
the histopathologic diagnosis of a given tumor. In fact,
in the current study, one RB⫺/p16⫹ tumor initially
diagnosed as TC was found to be an AC, and three
RB⫹/p16⫺ tumors initially diagnosed as SCLC were
found to be LCNECs by pathologic reevaluation (Table
1). In addition, one RB⫹/p16⫹ tumor initially diagnosed as AC was found to be a TC (Table 1). If RB⫹
ACs are found to be negative for p16 expression in
future studies, this may also allow a distinction between TCs and ACs to be made using this criterion. In
any case, more studies with larger cohorts of NE tumors of the lung will be needed to conclude that RB
and p16 protein status can be of definite use in histologic classification, because we analyzed small numbers of ACs and LCNECs in the current study.
Although NE tumors of the lung share common
phenotypic features, suggesting a genotypic relation,
they differ remarkably in their molecular genetic and
cytogenetic characteristics, including loss of RB and
p16 proteins,9,11,12,17,20,28 p53 mutations,9,10,12,14,16,17,22
MEN1 gene mutations,18 loss of heterozygosity (LOH)
of 3p alleles,15,19,22 and 11q deletion by comparative
genomic hybridization,21 highlighting an early fundamental molecular genetic divergence during the development of these tumors. The significant association between specific NE tumor types, including AC,
LCNEC, and SCLC, and heavy smoking with more than
20 pack-years; and the finding that patients with ACs,
LCNECs, and SCLCs were older than those with TCs in
this study suggest that development of these types of
NE lung tumors may be caused by alterations of the
genes, including the RB gene damaged by carcinogens
in tobacco smoke in older individuals after certain
periods of carcinogen exposure.
In conclusion, the current study indicates that
the p16:RB pathway of tumor suppression can be
altered by loss of either RB or p16 protein in ACs and
LCNECs and by loss of RB protein in SCLCs, whereas
the p16:RB pathway is normal in TCs. The RB and
p16 protein status determined by immunohistochemistry may also be useful as a diagnostic marker
for certain cases of LCNECs versus SCLCs and for
ACs versus TCs.
555
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Warren WH, Faber LP, Gould VE. Neuroendocrine neoplasms of the lung. J Thorac Cardiovasc Surg 1989;98:321–32.
Colby TV, Koss MN, Travis WD. Carcinoid and other neuroendocrine tumors. In: Rosai J and Sobin LH, editors. Atlas
of tumor pathology. 3rd series, Fascicle 13. Tumors of the
lower respiratory tract. Washington, DC: Armed Forces Institute of Pathology, 1995;287–317.
Cagle PT. Tumors of the lung (excluding lymphoid tumors).
In: Thurlbeck WM, Churg AM, editors. Pathology of the lung.
2nd edition. New York: Thieme Medical Publishers, Inc.,
1995;437–552.
Linnoila RI. Spectrum of neuroendocrine differentiation in
lung cancer cell line featured by cytomorphology, markers,
and their corresponding tumors. J Cell Biochem (Suppl)
1996;24:9 –106.
Travis WD, Linnoila RI, Tsokos MG, Hitchcock CL, Cutler GB
Jr., Nieman L, et al. Neuroendocrine tumors of the lung with
proposed criteria for large-cell neuroendocrine carcinoma.
Am J Surg Pathol 1991;15:529 –53.
Colby TV, Koss MN, Travis WD. Small cell carcinoma and
large cell neuroendocrine carcinoma. In: Rosai J and Sobin
LH, editors. Atlas of tumor pathology. 3rd series, Fascicle 13.
Tumors of the lower respiratory tract. Washington, DC:
Armed Forces Institute of Pathology, 1995:235–57.
Travis WD, Gal AA, Colby TV, Klimstra DS, Falk R, Koss MN.
Reproducibility of neuroendocrine lung tumor classification. Hum Pathol 1998;29:272–9.
Travis WD, Rush W, Flieder DB, Falk R, Fleming MV, Gal AA,
et al. Survival analysis of 200 pulmonary neuroendocrine
tumors with clarification of criteria for atypical carcinoid
and its separation from typical carcinoid. Am J Surg Pathol
1998;22:934 – 44.
Barbareschi M, Girlando S, Mauri FA, Arrigoni G, Laurino L,
Palma PD, et al. Tumor suppressor gene products, proliferation, and differentiation markers in lung neuroendocrine
neoplasms. J Pathol 1992;166:343–50.
Lohmann DR, Fesseler B, Putz B, Reich U, Bohm J, Prauer H,
et al. Infrequent mutations of the p53 gene in pulmonary
carcinoid tumors. Cancer Res 1993;53:5797– 801.
Gouyer V, Gazzeri S, Brambilla E, Bolon I, Moro D, Perron P,
et al. Loss of heterozygosity at the RB locus correlates with
loss of RB protein in primary malignant neuroendocrine
lung carcinomas. Int J Cancer 1994;58:818 –24.
Lai S, Brauch H, Knutsen T, Johnson BE, Nau MM, Mitsudomi T, et al. Molecular genetic characterization of neuroendocrine lung cancer cell lines. Anticancer Res 1995;15:
225–32.
Jiang S, Kameya T, Sato Y, Yanase N, Yoshimura H, Kodama
T. Bcl-2 protein expression in lung cancer and close correlation with neuroendocrine differentiation. Am J Pathol
1996;148:837– 46.
Przygodzki RM, Finkelstein SD, Langer JC, Swalsky PA, Fishback N, Bakker A, et al. Analysis of p53, K-ras, and c-raf-1 in
pulmonary neuroendocrine tumors. Am J Pathol 1996;148:
1531– 41.
Hurr K, Kemp B, Silver SA, El-Naggar AK. Microsatellite
alteration at chromosome 3p loci in neuroendocrine and
nonneuroendocrine lung tumors. Am J Pathol 1996;149:613–
20.
Brambilla E, Negoescu A, Gazzeri S, Lantuejoul S, Moro D,
Brambilla C, et al. Apoptosis-related factors p53, Bc12, and
Bax in neuroendocrine lung tumors. Am J Pathol 1996;149:
1941–52.
556
CANCER February 1, 2000 / Volume 88 / Number 3
17. Rusch VW, Klimstra DS, Venkatraman ES. Molecular markers help characterize neuroendocrine lung tumors. Ann
Thorac Surg 1996;62:798 – 810.
18. Debelenko LV, Brambilla E, Agarwal SK, Swalwell JI, Kester
MB, Lubensky IA, et al. Identification of MEN1 gene mutations in sporadic carcinoid tumors of the lung. Hum Mol
Genet 1997;6:2285–90.
19. Kovatich AK, Friedland DM, Druck T, Hadaczek P, Huebner
K, Comis RL, et al. Molecular alterations to human chromosome 3p loci in neuroendocrine lung tumors. Cancer 1998;
83:1109 –17.
20. Gazzeri S, Valle VD, Chaussade L, Brambilla C, Larsen CJ,
Brambilla E. The human p19ARF protein encoded by the b
transcript of the p16INK4a gene is frequently lost in small cell
lung cancer. Cancer Res 1998;58:3926 –31.
21. Walch AK, Zitzelsberger HF, Aubele MM, Mattis AE, Bauchinger M, Candidus S, et al. Typical and atypical carcinoid
tumors of the lung are characterized by 11q deletions as
detected by comparative genomic hybridization. Am J
Pathol 1998;153:1089 –98.
22. Onuki N, Wistuba I, Travis WD, Virmani AK, Yashima K,
Brambilla E, et al. Genetic changes in the spectrum of neuroendocrine lung tumors. Cancer 1999;85:600 –7.
23. Weinberg RA. The retinoblastoma protein and cell cycle
control. Cell 1995;81:323–30.
24. Benedict WF, Xu H, Hu S, Takahashi R. Role of the retinoblastoma gene in the initiation and progression of human
cancer. J Clin Invest 1990;85:988 –93.
25. Xu H, Hu S, Benedict WF. Lack of nuclear RB protein staining in G0/middle G1 cells: correlation to changes in total RB
protein level. Oncogene 1991;6:1139 – 46.
26. Xu H, Hu S, Cagle PT, Moore GE, Benedict WF. Absence of
retinoblastoma protein expression in primary non-small cell
lung carcinomas. Cancer Res 1991;51:2735–9.
27. Zhang X, Xu H, Murakami Y, Sachse R, Yashima K, Hirohashi
S, et al. Deletion of chromosome 1q, mutations in retinoblastoma 1, and retinoblastoma protein state in human
hepatocellular carcinoma. Cancer Res 1994;54:4177– 82.
28. Cagle PT, El-Naggar AK, Xu H, Hu S, Benedict WF. Differential retinoblastoma protein expression in neuroendocrine
tumors of the lung. Am J Pathol 1997;150:393– 400.
29. Kamb A, Gruis NA, Weaver-Feldhaus J, Liu Q, Harshman K,
Tavtigian SV, et al. A cell cycle regulator potentially involved
in genesis of many tumor types. Science 1994;264:436 – 40.
30. Nobori T, Miura K, Wu DJ, Lois A, Takabayashi K, Carson
DA. Deletions of the cyclin-dependent kinase-4 inhibitor
gene in multiple human cancers. Nature 1994;368:753– 6.
31. Serrano M, Hannon GJ, Beach D. A new regulatory motif in
cell-cycle control causing specific inhibition of cyclin
D/CDK4. Nature 1993;366:704 –7.
32. Lukas J, Parry D, Aagaard L, Mann DJ, Bartkova J, Strauss M,
et al. Retinoblastoma protein-dependent cell-cycle inhibition by the tumor suppressor p16. Nature 1995;375:503– 6.
33. Otterson GA, Kratzke RA, Coxon A, Kim YW, Kaye FJ. Absence of p16INK4 protein is restricted to the subset of lung
cancer cell line that retains wild type RB. Oncogene 1994;9:
3375– 8.
34. Shapiro GI, Edwards CD, Kobzik L, Godleski J, Richards W,
Sugarbaker DJ, et al. Reciprocal Rb inactivation and p16INK4
expression in primary lung cancers and cell lines. Cancer Res
1995;55:505–9.
35. Kelley MJ, Nakagawa K, Steinberg SM, Mulshine JL, Kamb A,
Johnson BE. Differential inactivation of CDKN2 and Rb protein in non-small-cell and small-cell lung cancer cell lines.
J Natl Cancer Inst 1995;87:756 – 61.
36. Geradts J, Kratzke RA, Niehans GA, Lincoln CE. Immunohistochemical detection of the cyclin-dependent kinase inhibitor 2/multiple tumor suppressor gene 1 (CDKN2/MTS1)
product p16INK4A in archival human solid tumors: correlation with retinoblastoma protein expression. Cancer Res
1995;55:6006 –11.
37. Kinoshita I, Dosaka-Akita H, Mishina T, Akie K, Nishi M,
Hiroumi H, et al. Altered p16INK4 and retinoblastoma protein status in non-small cell lung cancer: potential synergistic effect with altered p53 protein on proliferative activity.
Cancer Res 1996;56:5557– 62.
38. Benedict WF, Lerner SP, Zhou J, Shen X, Tokunaga H, Czerniak B. Level of retinoblastoma protein expression correlates with p16 (MTS-1/INK4A/CDKN2) status in bladder
cancer. Oncogene 1999;18:1197–203.
39. Logothetis CJ, Xu H, Ro JY, Hu S, Sahin A, Ordonez N, et al.
Altered expression of retinoblastoma protein and known
prognostic variables in locally advanced bladder cancer.
J Natl Cancer Inst 1992;84:1256 – 61.
40. Xu H, Hu S, Hashimoto T, Takahashi R, Benedict WF. The
retinoblastoma susceptibility gene product: a characteristic
pattern in normal cells and abnormal expression in malignant cells. Oncogene 1989;4:807–21.
41. Xu H, Quinlan DC, Davidson AG, Hu S, Summers CL, Li J, et
al. Altered retinoblastoma protein expression and prognosis
in early-stage non-small-cell lung carcinoma. J Natl Cancer
Inst 1994;86:695–9.
42. Mehta CR, Patel NR. A network algorithm for performing
Fisher’s exact test in r ⫻ c contingency tables. J Am Stat
Assoc 1983;78:427–34.
43. Arrigoni MG, Woolner LB, Bernatz PE. Atypical carcinoid
tumors of the lung. J Thorac Cardiovasc Surg 1972;64:
413–21.
Документ
Категория
Без категории
Просмотров
4
Размер файла
277 Кб
Теги
786
1/--страниц
Пожаловаться на содержимое документа