[No abstract available]sem informação843393397FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2011/23204-5; 2011/23366-

Altemani, A.

Del Negro, A.

Giovanetti, K.

Kowalski, L.P.

Krepischi, A.C.V.

Mariano, F.V.

Saccomani, L.F.V.

Brazilian Journal of Otorhinolaryngology

English

Repositorio da Producao Cientifica e Intelectual da Unicamp

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Braz J Otorhinolaryngol. 2016;xxx(xx):xxx--xxx
www.bjorl.org
Brazilian Journal of
OTORHINOLARYNGOLOGY
CASE REPORT
Genomic  profile  of  a squamous  cell  carcinoma  ex
pleomorphic adenoma  compared  to  a head  and neck
squamous cell  carcinoma
Perfil  genômico  de  um  carcinoma  espinocelular  ex-adenoma  pleomórfico
comparado  a  um  carcinoma  espinocelular  de  cabec¸a e  pescoc¸o
Fernanda Viviane Marianoa,∗, Luis Fernando Vidal Saccomania, Karina Giovanetti a,
André  Del Negrob, Luiz Paulo Kowalski c, Ana Cristina Victorino Krepischid,
Albina Altemania
a Pathology  Department,  Faculdade  de  Ciências  Médicas,  Universidade  Estadual  de  Campinas  (UNICAMP),  Campinas,  SP,  Brazil
b Head  and  Neck  Surgery,  Faculdade  de  Ciências  Médicas,  Universidade  Estadual  de  Campinas  (UNICAMP),  Campinas,  SP,  Brazil
c Head  and  Neck  Surgery  Department,  AC  Camargo  Cancer  Center,  São  Paulo,  SP,  Brazil
d Genetics  and  Evolutionary  Biology  Department,  Instituto  de  Biociências,  Universidade  de  São  Paulo  (USP),  São  Paulo,  SP,  BrazilReceived 6  October  2015;  accepted  13  November  2015
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hIntroduction
Squamous  cell  carcinoma  (SCC)  is  the  most  frequent  neo-
plasm  in  the  head  and  neck  and  in  oral  sites.  Head  and
neck  squamous  cell  carcinoma  (HNSCC)  is  the  sixth  most
common  solid  cancer,  affecting  more  than  900,000  peo-
ple  worldwide  each  year.  Oral  squamous  cell  carcinoma
(OSCC)  is  the  largest  subgroup  of  HNSCC  and  causes  7400
deaths  in  the  United  States  every  year.1 Because  of  the
 Please cite this article as: Mariano FV, Saccomani LFV, Giovanetti
K, Del Negro A, Kowalski LP, Krepischi ACV, et al. Genomic profile of
a squamous cell carcinoma ex pleomorphic adenoma compared to
a head and neck squamous cell carcinoma. Braz J Otorhinolaryngol.
2016. http://dx.doi.org/10.1016/j.bjorl.2015.11.015
∗ Corresponding author.
E-mail: fevimariano@gmail.com (F.V. Mariano).
m
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c
http://dx.doi.org/10.1016/j.bjorl.2015.11.015
1808-8694/© 2016 Associac¸ão Brasileira de Otorrinolaringologia e Cirurg
reserved.igh  prevalence  and  aggressiveness,  HNSCC  has  been  studied
y  several  molecular  and  cytogenetic  techniques.  Frequent
hromosomal  imbalances  reported  in  HNSCC  are  3p,  4,  5q,
p,  9p,  11,  13q,  and  18q  (losses)  and  3q25--26,  5p,  8q24,
q22--34,  11q13,  14q24,  16p,  19p,  20q24,  and  22q  (gains).1
The  carcinomatous  component  of  carcinoma  ex  pleomor-
hic  adenoma  (CXPA)  encompasses  a  variety  of  histological
umor  types,  such  as  adenocarcinoma,  myoepithelial  car-
inoma,  epithelial--myoepithelial  carcinoma,  sarcomatoid
arcinoma,  and  salivary  duct  carcinoma.2 Many  epidemio-
ogical  studies  about  incidence  of  CXPA  showed  few  or  no
ases  with  an  SCC  component.
Despite  reports  of  numerous  chromosomal  alterations  in
CC  from  other  sites,  no  particular  chromosomal  imbalances
ave  been  described  for  squamous  cell  carcinoma  ex  pleo-
orphic  adenoma  (SCC-CXPA).  The  aim  of  this  study  was
o  identify  chromosomal  changes  associated  with  SCC-CXPA,
omparing  with  HNSCC  data  from  the  literature.
ia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights
BJORL-313; No. of Pages 5
ARTICLE IN PRESS+Model
2  Mariano  FV  et  al.
Figure  1  Squamous  cell  carcinoma  ex  pleomorphic  adenoma.  (A,  B)  Residual  pleomorphic  adenoma.  (A)  Pleomorphic  adenoma
area shows  extensive  hyalinization  of  the  stroma.  (B)  Few  ductal  structures  lined  by  cells  without  atypical  features  can  be  seen.  (C,
D cell  c
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t) Carcinomatous  component.  (C)  A  frankly  invasive  squamous  
f the  malignant  component  showing  islands  formed  by  cells  wi
ase report
linicopathologic  data
he  case  was  referred  to  this  department  for  histopatholog-
cal  review.  The  gender  and  age  are  unknown.  The  patient
resented  a  mass  of  the  parotid  gland  that  had  been  notice-
ble  for  ten  years.  The  tumor  was  excised  and  histological
xamination  showed  pleomorphic  adenoma  (PA)  areas  and
CC  arising  in  PA  (Fig.  1).  Tumor  cells  presented  keratin  pearl
ormation  and  invaded  the  adjacent  tissues  of  parotid  gland.
he  neoplasm  was  classified  as  a  frankly  invasive  SCC-CXPA.
he  patient  developed  distant  metastasis  in  the  lungs  but
he  follow-up  information  was  not  available.
rray  comparative  genomic  hybridization  (CGH)
nalysis
omatic  copy  number  alterations  detected  in  the  SCC-
XPA  sample  are  detailed  in  Table  1  as  well  as  affected
nown  cancer  genes  according  to  the  Sanger  Cancer
ene  Census  (https://www.sanger.ac.uk/research/projects/
ancergenome/census.html).
The  current  case  exhibited  a  complex  genomic  pattern
ith  several  copy  number  alterations.  Losses  were  detected
t  1p,  8p,  17p,  and  18q,  and  gains  were  identified  at  1p,  6,
q,  11q,  17q,  19q,  and  22q  (Fig.  2A).
Three  regions  exhibiting  high  copy  number  alter-
tions  (amplifications)  were  detected  at  11q14.3q22.1,
i
t
s
carcinoma  is  the  malignant  component  of  the  tumor.  (D)  Detail
uamous  differentiation.
1q22.1q22.3  (Fig.  2B),  and  19p13.2  (Fig.  2C).  Among  other
enes,  these  amplifications  encompass  the  known  cancer
enes  MAML2,  BIRC3,  and  LYL1.
iscussion
ariano  et  al.2 reported  one  SCC-CXPA  out  of  38  cases
2.6%)  of  CXPA  from  a  Brazilian  population.  To  the  best  of
he  authors’  knowledge,  only  sporadic  cases  were  described
howing  SCC  as  the  malignant  component  of  CXPA,  empha-
izing  the  clinical  and  morphological  features.
SCC  originates  from  the  epithelial  component  of  the  PA,
nd  in  lesions  that  arise  from  minor  salivary  glands,  it  is
mportant  to  demonstrate  no  continuity  of  the  SCC  nests
ith  the  buccal  mucosa.  In  the  current  case,  areas  of  resid-
al  PA  were  associated  with  a  widely  invasive  SCC  in  the  soft
issues  surrounding  the  parotid  gland.  Distant  metastases
ere  observed  in  this  case,  suggesting  that  SCC  belongs  to
he  group  of  aggressive  CXPAs.  It  has  been  proposed  that
XPA  is  actually  a  category  of  tumors  rather  than  a  single
umor  type,  since  both  aggressive  and  indolent  carcinomas
an  arise  from  a PA.
Local  aggressiveness,  high  rate  of  early  relapse  tumors,
nd  reduced  five-year  survival  rates  not  exceeding  50%  are
ypical  clinical  features  of  SCC.  OSCC  causes  7400  deaths
n  the  USA  every  year.1 For  these  reasons,  during  the  last
wo  decades,  genome  screening  approaches  like  chromo-
omal  CGH  were  applied  to  disclose  the  molecular  basis  of
arcinogenesis  of  this  tumor.
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Genomic  profile  of  SCC  ex  pleomorphic  adenoma  compared  to  HNSCC  3
Table  1  Somatic  copy  number  alterations  detected  by  array  comparative  genomic  hybridization  in  one  squamous  cell  carcinoma
ex pleomorphic  adenoma  case.
Chromosome  coordinates
(Hg19)
Event  type  Size
(Mb)
Cytoband  Genes
(n)
Know  cancer  genes  (CGCS)
chr1:783,108-10,814,886  Gain  10.0  1p36.33--p36.22  162  TNFRSF14,  PRDM16,  RPL22,
CAMTA1
chr1:55,751,323-74,536,392  Loss  18.7  1p32.3--p31.1  94  JUN,  JAK1
chr1:106,671,417-121,354,529  Loss  14.6  1p21.1--p11.2  171  RBM15,  TRIM33,  NRAS,  FAM46C,
NOTCH2
chr6:122,468-21,138,924  Gain  21.0  6p25.3--p22.3  126  IRF4,  DEK
chr6:147,333,160-170,923,962  Gain  23.5  6q24.3--q27  154  EZR,  FGFR1OP,  MLLT4
chr8:239,745-43,440,193  Loss  43.0  8p23.3--p11.1  364  PCM1,  WRN,  WHSC1L1,  FGFR1,
HOOK3
chr8:93,473,949-109,349,316  Gain  15.8  8q22.1--q23.1  94  COX6C
chr11:84,971,351-92,399,403  Gain  7.4  11q14.1--q14.3  43  PICALM
chr11:92,410,361-99,634,930  Amplificationa 7.0  11q14.3--q22.1  48  MAML2
chr11:99,645,163-104,334,763  Amplificationa 4.6  11q22.1--q22.3  29  BIRC3
chr17:9,134,319-22,224,426  Loss  13.0  17p13.1--p11.1  165  GAS7,  MAP2K4
chr17:46,160,506-59,942,017  Gain  13.7  17q21.32--q23.2  177  COL1A1,  HLF,  MSI2,  CLTC,  BRIP1
chr18:18,440,032-78,062,375  Loss  59.6  18q11.1--q23  278  ZNF521,  SS18,  MALT1,  BCL2
chr19:12,951,065-13,396,186  Amplificationa 0.4  19p13.2  19  LYL1
chr19:28,039,051-59,128,983  Gain  31.0  19q11--q13.43  1123  CCNE1,  CEBPA,  AKT2,  CD79A,
CIC,  BCL3,  CBLC,  ERCC2,  KLK2,
PPP2R1A,  ZNF331,  TFPT
chr22:31,865,770-43,582,078  Gain  11.7  22q12.2--q13.2  205  MYH9,  PDGFB,  MKL1,  EP300
a High copy number gains.
Genomic profile of SCNA in case 1
Chromosome 1p to Xq
A
B C
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Figure  2  Copy  number  alterations  detected  by  array  comparative  genomic  hybridization  (CGH)  in  the  squamous  cell  carcinoma
ex pleomorphic  adenoma  case.  (A)  Array-CGH  genomic  profile  exhibiting  the  all  the  identified  copy  number  alterations  of  this  case.
The x-axis  represents  probes  ordered  according  to  their  genomic  position  from  chromosomes  1p  to  Xq  (each  chromosome  is  labeled
with a  different  color).  The  y-axis  denotes  the  log2 test/reference  values  (genomic  gains  and  losses  are  plotted  above  or  below
the 0  baseline,  respectively;  images  adapted  from  the  software  Nexus  Copy  Number  7.0,  Biodiscovery).  The  red  box  indicates
chromosomes  that  harbor  genomic  rearrangements  of  losses  or  gains:  1,  6,  8,  11,  17,  18,  19  and  22.  The  arrows  show  the  regions  of
genomic amplifications  at  chromosomes  11  and  19.  (B)  Array-CGH  profile  of  the  chromosome  11  showing  in  detail  the  two  regions
with high  copy  number  gain  (amplifications):  at  11q14q22.1  (7  Mb)  and  at  11q22.1-q22.3  (4.6  Mb)  (blue  arrow  in  A).  (C)  Array-CGH
profile of  the  chromosome  19  exhibiting  one  region  of  genomic  amplification  at  19p13.2  (0.4  Mb)  (red  arrow  in  A).
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3ARTICLE
 
A  complex  genomic  profile  of  the  copy  number  alterations
f  HNSCC  has  been  described  in  the  literature,  including
p,  4,  5q,  8p,  9p,  11,  13q,  and  18q  losses,  and  3q25q26,
p,  8q24,  9q22q34,  11q13,  14q24,  16p,  19q,  20q24,  and
2q  gains.1 The  current  case  also  shows  a  complex  genomic
rofile  with  several  copy  number  alterations,  but  with  a
ifferent  pattern  compared  to  SCC  from  head  and  neck,
robably  due  to  distinct  cellular  lineage.  Interestingly,  in
NSCC  there  are  multiple  discrete  regions  of  loss  at  3p.1,3
ven  though  cytogenetic  investigations  have  suggested  that
p  loss  is  a  key  genomic  change  in  HNSCC,  the  present
ase  did  not  show  this  alteration.  However,  some  similar-
ties  between  HNSCC  and  this  case  were  found,  such  as  8p23
nd  18q11  losses,  and  8q22.1,  19q,  and  22q  gains.  These
lterations  can  be  specific  to  squamous  cell  differentiation
nd  contribute  to  aggressive  behavior.3
It  has  been  suggested  that  in  HNSCC,  gains  on  3q26  and
1q13  and  deletions  on  8p23  and  22q  could  be  valuable
arkers  of  aggressive  disease.1 The  loss  at  8p23  in  the
urrent  case  encompasses  important  cancer-related  genes,
uch  as  PCM1,  WRN,  WHSCILI,  FGFR1,  and  HOOK3.  However,
hromosomal  gains  on  1q  and  16q  as  well  as  chromosomal
oss  on  18q  have  been  suggested  to  represent  prognostic
arkers  in  HNSCC.1,3 The  present  report  also  found  loss  at
8q  involving  the  cancer  related  genes  ZNF521,  SS18,  MALT1,
nd  BCL2.  A  gain  at  8q22.1q23.1  was  also  observed.  In  this
enomic  interval,  the  YWHAZ  gene  is  mapped,  a  HNSCC
roto-oncogene  candidate.
New  findings  not  previously  reported  for  HNSCC  were
lso  identified,  such  as  losses  at  1p32,  1p21,  and  17p13.1,
s  well  as  gains  at  1p36.33,  6p25.3,  6q24,  11q14,  and
7q21.32.  The  1p32  loss  is  associated  with  the  develop-
ent  of  adenoid  cystic  carcinoma,1 and  1p36  is  frequently
eleted  in  human  cancers.  Among  these  regions  of  imbal-
nce,  1p21.1  loss  seems  particularly  interesting,  because  it
omprises  several  cancer  related  genes.1 Overexpression  of
he  DEK  gene  (6p25.3)  is  associated  with  gastric  and  breast
denocarcinoma,4 as  well  as  the  role  EZRIN  has  in  colorectal,
ancreatic,  gastric,  and  other  adenocarcinomas.5
It  was  noteworthy  that  three  genomic  amplifications
ere  detected  encompassing  the  region  at  11q14.3q22.1,
1q22.1q22.3,  and  19p13.2.  These  amplifications  can  har-
or  relevant  genes  driving  the  carcinogenesis,  and  MAML2,
IRC3,  and  LYL1  are  known  cancer  genes  mapped  within
hese  regions.
MAML2  (11q14.3q22.1)  (mastermind-like  2)  has  a role  as
 transcriptional  coactivator  for  the  Notch  receptor,  and
as  reported  to  be  rearranged  with  CRTC1  and  CRTC3  in
ucoepidermoid  carcinomas.6 In  the  present  case,  the  diag-
osis  of  mucoepidermoid  carcinoma  arising  in  PA  was  ruled
ut  in  the  histologic  examination  since  the  malignant  com-
onent  did  not  contain  mucous  cells.  MAML2-CRTC1  and
AML2-CRTC3  have  not  been  found  in  adenosquamous  carci-
oma,  SCC,  or  any  other  salivary  gland  carcinoma.6 MAML2
mplified  induces  the  transcription  of  the  HES-1  gene  (a
otch  target  gene)  that  regulates  the  expression  of  tissue-
pecific  transcription  factors  that  influence  lineage  and
ther  events.  This  amplification  is  described  with  a  low  fre-
uency  in  cancers  of  several  sites,  but  not  in  HNSCC.
Another  amplified  gene,  BIRC3  (baculoviral  IAP  repeat
ontaining  3),  encodes  a  member  of  the  IAP  family  of  pro-
eins  that  inhibit  apoptosis.  Amplification  involving  BIRC3
4 PRESS
Mariano  FV  et  al.
as  been  detected  in  pancreatic  cancer,  osteosarcoma,
cute  myeloid  leukemia,  mammary  carcinoma,  lung  can-
er,  cervical  cancer,  and  SCC.7 It  has  been  found  that
IRC3  may  enhance  cancer  cell  survival  and  cancer  progres-
ion,  suggesting  that  treatments  targeting  this  gene  product
ay  provide  useful  therapeutic  options.  LYL1  (lymphoblastic
eukemia-associated  hematopoiesis  regulator  1)  gene  rep-
esents  a basic  helix-loop-helix  transcription  factor,  which
ncodes  a  protein  with  roles  in  blood  vessel  maturation  and
ematopoiesis.8 It  is  believed  that  in  salivary  gland  tumors
YL1  amplification  could  contribute  with  oncogenesis.
PA  as  well  as  CXPA  has  been  reported  to  harbor
ighly  specific  chromosome  abnormalities  related  to  PLAG1
nd  HMGA2  genes.  Rearrangements  of  these  genes  (8q12
earrangements,  target  gene  PLAG1  and  12q14-15  rear-
angements,  target  gene  HMGA2)  have  been  implicated  in
he  genesis  of  PA  and  CXPA.  However,  although  these  chro-
osome  abnormalities  are  frequent  in  these  tumors,  they
re  not  detected  in  all  of  them.9 In  the  current  case,  ampli-
ed  PLAG1  or  HMGA2  genes  were  not  found,  suggesting  that
he  tumor  probably  belonged  to  the  group  without  such  kary-
tic  alteration.
onclusion
n  summary,  SCC-CXPA  showed  a  complex  pattern  of  copy
umber  alterations  that  has  some  similarities  with  that  of
SCCN.  It  is  likely  that  these  chromosomal  abnormalities
ay  be  involved  in  the  development  of  the  squamous  phe-
otype  and  aggressive  behavior  of  the  SCC-CXPA.  However,
enomic  amplifications  of  known  cancer  genes,  such  as  LYL1,
IRC3,  and  MAML2  detected  only  in  SCC-CXPA  could  have
mportance  for  the  process  of  malignant  transformation  of
A.  These  latter  alterations  may  represent  clinically  relevant
arkers  for  SCC-CXPA  diagnosis.
unding
APESP  Process  Nos.  2011/23204-5  and  2011/23366-5.
onflicts of  interest
he  authors  declare  no  conflicts  of  interest.
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http://dx.doi.org/10.1097/PAI.0b013e3182936ea7.


Genomic profile of a squamous cell carcinoma Ex pleomorphic adenoma compared to a head and neck squamous cell carcinoma

http://repositorio.unicamp.br/jspui/bitstream/REPOSIP/319735/1/2-s2.0-84959898603.pdf

Genomic profile of a squamous cell carcinoma Ex pleomorphic adenoma compared to a head and neck squamous cell carcinoma

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