Human Pathology (2014) 45, 2430–2436

www.elsevier.com/locate/humpath

Original contribution

Immunohistochemical detection of ARID1A in

colorectal carcinoma: loss of staining is associated
with sporadic microsatellite unstable tumors with
medullary histology and high TNM stage☆,☆☆
Jiqing Ye MD a , Yi Zhou MD a , Martin R. Weiser MD b , Mithat Gönen PhD c ,
Liying Zhang MD a , Tushar Samdani MD b , Ruben Bacares BS a , Deborah DeLair MD a ,
Sinisa Ivelja MD a , Efsevia Vakiani MD, PhD a , David S. Klimstra MD a ,
Robert A. Soslow MD a , Jinru Shia MD a,⁎
a
Department of Pathology, Memorial Sloan–Kettering Cancer Center, 12705 York Ave, New York, NY, 10065
b
Department of Surgery, Memorial Sloan–Kettering Cancer Center, 12705 York Ave, New York, NY, 10065
c
Department of Biostatistics, Memorial Sloan–Kettering Cancer Center, 12705 York Ave, New York, NY, 10065

Received 3 June 2014; revised 8 August 2014; accepted 13 August 2014

Keywords:
Summary AT-rich interactive domain-containing protein 1A (ARID1A), a chromatin remodeling gene
Colon cancer;
recently discovered to be a tumor suppressor in ovarian cancers, has been found to be mutated at low
Microsatellite instability;
frequencies in many other tumors including colorectal carcinoma (CRC). An association between ARID1A
Immunohistochemistry;
alteration and DNA mismatch repair (MMR) deficiency has been implicated; understanding this association
MLH1 and PMS2;
may facilitate the understanding of the role of ARID1A in the various tumors. In this pilot study, we analyzed
MLH1 methylation
the immunohistochemical expression of ARID1A in a consecutive series of 257 CRCs that fulfilled a set of
relaxed criteria for Lynch syndrome screening; 59 (23%) were MMR deficient by immunohistochemistry
(44 MLH1/PMS2 deficient, 9 MSH2/MSH6 deficient, 4 MSH6 deficient, and 2 PMS2 deficient). ARID1A
loss was observed in 9% (22/257) of the cohort: 24% of MMR-deficient tumors (14/59, 13 of the 14 being
MLH1/PMS2 deficient) and 4% of MMR-normal tumors (8/198) (P b .05). MLH1 (mutL homolog 1)
promoter hypermethylation was observed in 10 of the 13 MLH1/PMS2-deficient/ARID1A-loss tumors,
indicating an association between ARID1A loss and sporadic microsatellite unstable CRCs. Among the
MMR-deficient cases, ARID1A loss correlated with old age (P = .04), poor tumor differentiation (P b .01),
medullary histology (P b .01), and an increased rate of nodal and distant metastasis (P = .03); these patients
also trended toward a worse 5-year overall survival. Among MMR-normal tumors, no differences in
clinicopathological features were detected between the groups stratified by ARID1A. In conclusion, our
results suggest that ARID1A loss may be linked to a specific subset of sporadic microsatellite unstable CRCs
that may be medullary but is more likely to present with metastatic disease, warranting further investigation.
© 2014 Elsevier Inc. All rights reserved.

☆
This study was presented in part at the 102nd annual meeting of the United States and Canadian Academy of Pathology in Baltimore, MD, in March 2013.
☆☆
Disclosures: The authors declare no conflict of interest.
⁎ Corresponding author.
E-mail address: Shiaj@mskcc.org (J. Shia).

http://dx.doi.org/10.1016/j.humpath.2014.08.007
0046-8177/© 2014 Elsevier Inc. All rights reserved.
ARID1A in colorectal carcinoma
1. Introduction
AT-rich interactive domain-containing protein 1A
(ARID1A) (also known as BAF250a), encoded by the AR-
ID1A gene, is a key component of the large adenosine
triphosphate–dependent chromatin remodeling complex
SWItch/Sucrose NonFermentable (SWI/SNF) [1]. Owing
to the advanced sequencing techniques, this gene was first
found to be mutated frequently in certain carcinomas,
specifically, ovarian clear cell [2,3] and endometrioid [3]
carcinomas and uterine endometrioid carcinomas [4]. In these
tumors, ARID1A is believed to act as a tumor suppressor gene
[2-6]. Subsequently, mutations in this gene have been found to
occur in small subsets of carcinomas from a wide variety of
organ sites, including colon and rectum, stomach, pancreas,
liver, prostate, breast, and lung [7].
Studies have suggested that, in at least some tumor types,
ARID1A mutation is associated with microsatellite instability
(MSI) [7-9]. Jones et al [7] reported that, among gastric, colon,
prostate, and pancreatic carcinomas, the most commonly
observed mutations in ARID1A occurred in a coding
microsatellite, specifically, a 7-base G tract in the coding
region; and 12 of 12 MSI-high (MSI-H) cancers (6 colon, 5
gastric, and 1 prostate) that had ARID1A mutation had their
mutations at mononucleotide tracts in the gene. Interestingly, a
recent study [6] indicated that, in the case of endometrial
carcinoma, loss of ARID1A protein was significantly more
prevalent in sporadic MSI-H cancers with MLH1 promoter
hypermethylation than in Lynch-associated tumors (75%
versus 14%, P b .0001). Such developments have prompted
the speculation that ARID1A mutation may drive endometrial
carcinogenesis toward MSI through epigenetic alterations of
the MLH1 gene.
Mutation of ARID1A has been identified as a recurrent event
in colorectal carcinoma (CRC) in the recent Cancer Genome
Atlas project (TCGA) data [10]. Its specific role in colorectal
tumor development and progression, however, remains to be
determined. Although it has been suggested that an association
between ARID1A alteration and MSI might exist in CRC as
well [7,10], this has not been systematically studied. Thus, in this
pilot study, we evaluated the expression of ARID1A in a series
of CRCs stratified by the expression status of DNA mismatch
repair (MMR) proteins and MLH1 promoter methylation in
selected cases and explored the clinicopathological characteris-
tics associated with abnormal tumor expression of ARID1A.
Our study provided interesting findings that may serve to guide
further investigations that aim at understanding the biological
and clinical significance of ARID1A alteration in CRC.
2. Materials and methods
2.1. Patient information
Study cases were identified from a consecutive series of
CRC patients who fulfilled at least one of the following
2431
criteria: (1) age 50 years or younger, (2) age older than
50 years and the tumor showed MSI-H histology as defined
by the Revised Bethesda guidelines [11], and (3) family and/or
personal history meeting the Revised Bethesda guidelines as
determined by the treating physician. The patients were treated
at Memorial Sloan–Kettering Cancer Center from 2007 to
2009. During this period, colorectal cancers from patients
fulfilling any 1 of the above 3 criteria were routinely tested
with immunohistochemical stains for MLH1, MSH2, MSH6,
and PMS2. The patients' clinical information was retrieved
from the hospital information system. The study was approved
by the institutional board review.
2.2. Tumor pathology
All study cases were systematically reviewed for the
following pathologic features: (1) tumor location (right
colon referring to cecum, ascending and transverse colon,
and left colon referring to descending and sigmoid colon)
and (2) histologic grade (low grade referring to well or
moderately differentiated adenocarcinoma and high grade
referring to poorly differentiated adenocarcinoma with
b50% of the tumor showing gland formation) [12].
Medullary histology represented by solid growth with
prominent intratumoral and stromal lymphocytes [13] was
considered to be “high grade,” its presence in greater than or
equal to 25% of the tumor was recorded as “medullary
component present,” and its presence in greater than or
equal to 50% of the tumor qualified the tumor for
“medullary carcinoma”; (3) tumor-infiltrating lymphocytes
(TILs) (count per 10 high-power fields); and (4) tumor stage
(as defined by the seventh edition of American Joint
Committee on Cancer Cancer Staging Manual [12]).
2.3. Tissue microarray
Representative tumor blocks were selected from each of the
study cases and used for construction of tissue microarray
(TMA). The ATA-27 automated arrayer (Beecher Instruments,
Sun Prairie, WI, USA) was used to create the microarrays. The
punch size was set at 0.6 mm. Three cores of different areas of
the tumor were sampled from each tumor specimen.
2.4. Immunohistochemistry
Immunohistochemistry (IHC) for MLH1, PMS2, MSH2,
and MSH6 was performed on whole tumor sections of each
patient's resection specimen, using standard streptavidin-
biotin-peroxidase procedure. Primary monoclonal antibodies
used included MLH1 (clone G168-728, diluted 1:250; BD
PharMingen, San Diego, CA), MSH2 (clone FE11, diluted
1:50; Oncogene Research Products, Cambridge, MA), MSH6
(clone GRBP.P1/2.D4, diluted 1:200; Serotec, Raleigh, NC),
and PMS2 (clone A16-4, diluted 1:200; BD PharMingen).
Nonneoplastic colonic mucosa and colorectal tumors known
2432
to be deficient of MLH1, MSH2, MSH6, and PMS2 were
used as external positive and negative controls, respectively.
Abnormal MMR IHC was defined as complete absence of
nuclear staining within tumor cells in the presence of
concurrent positive labeling in internal nonneoplastic tissues.
IHC for ARID1A was performed on the TMA slides using a
polyclonal antibody against ARID1A (HPA005456; Sigma-
Aldrich, St. Louis, MO, USA) [4] and on the Ventana, Tucson,
AZ, USA Discovery XT. The concentration of the polyclonal
antibody was 0.2 mg/mL. The dilution factor of this antibody
was optimized by using liver parenchyma as negative control
and tonsil tissue as positive control as recommended. In addition
to antibody concentration, we also optimized target retrieval
buffer. After optimizing the IHC conditions, the TMA slides
were deparaffinized. Target retrieval was performed in standard
CC1 slightly basic buffer. The primary antibody was diluted by
1:100 with a final concentration of 0.2 μg/mL. The incubation
time for the primary antibody was 60 minutes. The slides were
washed, followed by incubation with the secondary antibody
and final development with the 3,3′-Diaminobenzidine Map Kit
(Ventana, catalog no. 750-169) for 60 minutes. Abnormal
ARID1A IHC was defined as complete loss of nuclear staining in
the tumor. Conversely, normal ARID1A IHC included cases
with positive nuclear staining, including cases with weak
staining (intensity comparable with stromal cells) and strong
staining (intensity stronger than stromal cells). Cases with no
analyzable core in the TMA were excluded from final analysis.
The minimal requirement for a case to be regarded as analyzable
was the presence of at least 1 intact core.
2.5. MLH1 promoter methylation
For each case, a 5-micron section of formalin-fixed,
paraffin-embedded tumor tissue was stained by hematoxylin
and eosin to identify areas of tumor on the slide. Dissection
of the tumor area was carried out from the corresponding
unstained sections. The tumor tissue was deparaffinized, and
DNA extraction was performed using the Qiagen DNeasy
Tissue Kit (cat no. 69504; Qiagen, Valencia, CA, USA).
Tumor DNA was amplified to test for the presence of
hypermethylation of 5 CpG sites within the MLH1 promoter
(in region −209 to −181 from the transcription start site). The
polymerase chain reaction product was subject to pyrose-
quencing on a Pyromark Q24 pyrosequencer. Because
bisulfite treatment converts unmethylated Cs to Ts, the
degree of methylation for each CpG site was calculated as
follows: methylation % = peak height of methylated (C)/
(peak height of methylated [C] + peak height nonmethylated
[T]) × 100. Methylation status was graded as “present,” when
all 5 CpG sites were methylated above 10%.
2.6. Statistical analysis
Cases were categorized into subgroups based on staining
results for MMR proteins and ARID1A. The frequencies of
J. Ye et al.
the various subgroups were documented. The clinical and
pathologic features were comparatively analyzed when
feasible. Continuous data were described as mean and SE
or median and range as appropriate. Categorical data were
described as numbers and percentages. The significance of
differences in categorical variables between groups was
tested by the Fisher exact tests. Survival probabilities were
estimated by the method of Kaplan-Meier and compared
using the log-rank test. Statistical significance was defined
as P b .05.
3. Results
A total of 302 CRCs from 301 patients were included in the
TMA. Of them, 45 cases (15%) were excluded due to tissue
loss, leaving 257 analyzable cases obtained from 256 patients.
Of the 257 tumors, 59 (23%) had abnormal MMR IHC
(Table 1). The abnormal patterns were as follows: 44 tumors
losing MLH1/PMS2, 2 tumors losing PMS2 alone, 9 tumors
losing MSH2/MSH6, and 4 tumors losing MSH6 alone.
The staining for ARID1A was easily interpretable (Fig.).
Normal staining was detected in 235 cases (235/257, 91%);
55% of which (129/235) showed a strong intensity, whereas
the remaining 45% (106 cases) had a weaker but easily
discernible intensity. Abnormal ARID1A IHC defined as loss
of tumor cell staining was detected in 9% (22/257) of the
series; all abnormal cases had discernible positive internal
control. Some variation in staining intensity (moderate versus
strong or moderate versus weak) was noted in a small subset of
positively stained cases. Overall, we did not encounter
significant staining heterogeneity among different cores.
Abnormal ARID1A was significantly more prevalent in
the MMR-abnormal group than in the MMR-normal group
(14/59 or 24% versus 8/198 or 4%, P b .05). Conversely,
abnormal MMR was significantly more common in the
ARID1A-abnormal group than in the ARID1A-normal
group (14/22 or 64% versus 45/236 or 19%, P b .05).
Overall, a total of 14 cases (14/257, 5%) were abnormal in
both MMR and ARID1A; of the 14 cases, 13 (93%) were
Table 1 Number of colorectal carcinoma cases showing
normal versus abnormal immunohistochemical expression of
DNA MMR proteins and ARID1A a
ARID1A IHC b MMR IHC b
Abnormal Normal Total
Abnormal 14 (24%) 8 (4%) 22 (9%)
Normal 45 (76%) 190 (96%) 235 (91%)
Total 59 198 257
a
P b.05.
b
For IHC for both MMR proteins and ARID1A, “abnormal”
indicates no nuclear staining in the tumor cells, and “normal” indicates
the presence of nuclear staining in the tumor cells.
ARID1A in colorectal carcinoma 2433

Fig. Photomicrographs of 2 colonic adenocarcinomas with different ARID1A immunohistochemical staining results. One is a moderately
differentiated adenocarcinoma (A) and shows positive nuclear labeling for ARID1A in both the tumor cells and the background nonneoplastic
cells (B). The second tumor is a medullary carcinoma (C) and shows loss of ARID1A staining in the tumor cells, whereas the background
nonneoplastic cells are positively stained (D) (original magnification, hematoxylin and eosin ×100 [A-D]).

abnormal in MLH1/PMS2 with only 1 case showing a ed promoter hypermethylation (as defined in the Methods
different abnormal pattern (loss of MSH6 alone). In contrast, section). All 10 cases had loss of both MLH1 and PMS2 by
31% (14/45) of the MMR-abnormal/ARID1A-normal cases IHC. Three MLH1/PMS2-abnormal cases (aged 36, 65, and 79
showed an MMR abnormal pattern different from loss of years, respectively) and 1 MSH6-abnormal case (age 50 years)
MLH1/PMS2. This is outlined in Table 2. did not exhibit MLH1 promoter methylation.
MLH1 promoter methylation analysis was performed on the The clinicopathological features of the various subgroups
14 ARID1A-abnormal/MMR-abnormal cases; 10 demonstrat- of the cases are summarized in Table 3.
Among MMR-abnormal cases, the group that also had
ARID1A abnormality showed an average age 11 years
older than the group with normal ARID1A (mean, 69.6
Table 2 Patterns of MMR-protein abnormality in colorectal
carcinomas with and without abnormal ARID1A a versus 58.4 years; median, 73.5 versus 62 years, P = .04).
When compared with the group with normal ARID1A, the
Patterns of MMR ARID1A b ARID1A-abnormal group tended to have tumors with
IHC abnormality b Abnormal Normal Total high histologic grade (11/14 or 79% versus 10/44 or 23%,
Loss of MLH1/PMS2 13 (93%) 31 (69%) 44 (75%) P b .01) and of medullary type, that is, having a
Other patterns medullary component in greater than or equal to 50% of
Loss of PMS2 0 2 2 the tumor (8/14 or 43% versus 5/45 or 11%, P b .01),
Loss of MSH2/MSH6 0 9 9 presented at later TNM stages (stages III/IV, 7/14 or 50%
Loss of MSH6 1 3 4 versus 8/45 or 18%, P = .03), and exhibited a trend (not
Subtotal 1 (7%) 14 (31%) 15 (25%) statistically significant) toward a worse 5-year overall
Total no. of cases 14 45 59 survival (68% versus 80%, P = .23) with a median
a
P = .08. follow-up time of 57 months (range, 1-138 months). TILs
b
For IHC for both MMR proteins and ARID1A, “abnormal” were increased in both ARID1A-normal and abnormal
indicates no nuclear staining in the tumor cells, and “normal” indicates
groups with no significant difference in the numbers of
the presence of nuclear staining in the tumor cells.
TILs/10 high-power fields.
2434 J. Ye et al.

Table 3 Correlation between immunohistochemical expression of MMR protein and ARID1A and clinicopathological features a
MMR IHC abnormal MMR IHC normal
ARID1A ARID1A
Total, Abnormal, Normal, P Total, Abnormal, Normal, P
n = 59 n = 14 n = 45 n = 198 a n=8 n = 190
Age (y)
Mean 61.0 69.6 58.4 .04 47.7 48.8 47.6 .77
Median 64.0 73.5 62 47 49.5 47
Range 29-89 36-89 29-85 18-86 44-53 18-86
Sex (n)
Male 27 5 22 .54 105 b 5 100 .72
Female 32 9 23 93 3 90
Tumor location (n)
Right colon 50 12 38 .69 55 0 55 .11
Left colon 7 1 6 58 5 53
Rectum 2 1 1 85 3 82
Histology grade (n)
Low 37 3 34 b.01 179 8 171 1.00
High 21 11 10 14 0 14
NA 1 0 1 5 0 5
Medullary carcinoma c (n) 13 8 5 b.01 1 0 1 1.00
TIL (count per 10 HPFs)
Mean 200 211 191 .44 14 6 14 .43
Median 198 231 173 4 1 4
Range 0-680 6-560 0-680 0-247 0-30 0-247
Tumor stage (n)
I and II 44 7 37 .03 84 3 81 1.00
III and IV 15 7 8 114 5 109
5-Year overall survival rates (%) 77% 68% 80% .23 66% 83% 65% .87
Abbreviations: NA, not available; TIL, tumor-infiltrating lymphocytes; HPFs, high-power fields.
a
For IHC for both MMR proteins and ARID1A, “abnormal” indicates no nuclear staining in the tumor cells and “normal” indicates the presence of
nuclear staining in the tumor cells.
b
One patient had 2 synchronous cancers.
c
Defined as having greater than or equal to 50% of the tumor showing medullary pattern.

In contrast, among the MMR-normal cases, no apparent
difference was detected between the groups stratified by the
ARID1A status in any of the features examined (Table 3).
4. Discussion
This study analyzed a specific population of colorectal
cancer patients that was considered to be at increased risk for
Lynch syndrome based on the Bethesda guidelines. The rate
of MMR protein loss by IHC was 23%. In such a population,
we identified ARID1A loss (defined as loss of IHC staining
in the tumor cells) in 9% of the cases. Our results further
indicated that (1) there existed a tight association between
ARID1A loss and MMR deficiency in CRC, with most
ARID1A-abnormal tumors being MLH1/PMS2 deficient
and showing MLH1 promoter hypermethylation, although
notably, ARID1A loss was observed in an MSH6-deficient
tumor as well; and (2) among the cases with abnormal MMR,
ARID1A loss correlated with the following clinicopatholog-
ical features: old age (on average, approximately 10 years
older than ARID1A-normal cases); higher tumor histologic
grade; medullary histology; and a more advanced TNM
stage, with a trend (not reaching statistical significance)
toward a worse 5-year overall survival.
The incidence of ARID1A loss in colorectal cancers in
general remains to be determined. Our study population was
biased toward patients with increased risk for Lynch
syndrome; thus, the observed rate of 9% may not represent
the general colorectal cancer patient population. Interesting-
ly, however, this rate is very close to the 10% of colorectal
cancer cases showing somatic mutations in ARID1A reported
by Jones et al [7] and suggests that, although the subset of
colorectal cancers that has ARID1A abnormality might be
small, it is not negligible. Current concepts hold that driver
genes frequently present in 1 tumor entity often turn out to be
significant in small subsets of other tumor types as well.
Given that ARID1A is shown to be a driver gene that is
frequently altered in ovarian carcinomas [2], the observation
ARID1A in colorectal carcinoma
that it is abnormal in a small subset of colorectal cancers as
well suggests that this gene also bears the potential to be
important in these colorectal cancers. This is of value and
deserves our attention.
Our finding of a significantly higher frequency of
ARID1A loss in MMR-deficient CRCs than in MMR-
normal CRCs is in line with the published literature
[7,10,14]. Adding to the existing literature, we further
observed that the MMR-deficient CRCs that show ARID1A
loss are primarily deficient in MLH1/PMS2 and harbor
MLH1 promoter hypermethylation, that is, sporadic MSI
cancers, and these patients, not surprisingly, are on average
approximately 10 years older than patients with MMR-
abnormal/ARID1A-normal tumors. In contrast, of the MMR-
deficient/ARID1A-normal group, more than 30% of the
tumors were MMR deficient in patterns other than concurrent
loss of MLH1/PMS2, suggesting mechanisms other than
MLH1 promoter hypermethylation. Unfortunately, however,
detailed genetic data were not available.
A significant association between abnormal ARID1A and
sporadic MSI with MLH1 promoter methylation has indeed
been reported in endometrial cancers [6]. There, it has been
speculated that ARID1A loss might have resulted in
epigenetic alterations by a deficient SWI/SNF complex
with subsequent MLH1 promoter methylation, or that the
loss of both ARID1A and MLH1 may be a reflection of
global genomic hypermethylation, that is, the CpG island
methylator phenotype. This is a tenable hypothesis because
ARID1A has been shown to harbor CpG islands in its
promoter, and ARID1A promoter hypermethylation has indeed
been described in breast carcinoma in a recent report [15].
These hypotheses are likely applicable to colorectal tumors as
well given the findings of our study. However, they remain to
be verified. To this end, it is particularly worth noting that loss
of ARID1A protein has been observed to be associated with
mutations of the gene [4], which would conflict with the
mechanism of promoter methylation. This, coupled with our
finding of abnormal ARID1A in an MSH6-deficient/MLH1
methylation–negative tumor, suggests that ARID1A alteration
in tumors may be mediated by heterogeneous mechanisms.
Very interestingly, our results indicated that, among
MMR-deficient CRCs, those that were also ARID1A
abnormal not only had MLH1 promoter hypermethylation
and were from significantly older patients, they also had
other distinct pathologic features when compared with cases
with normal ARID1A. First, these ARID1A-abnormal
tumors were of higher histologic grade and more frequently
of medullary type. Second and more importantly, these cases
were more likely to present at an advanced TNM stage.
These findings are of interest and engender new speculations
with regard to the prognostic implications of medullary
histology, MSI, and ARID1A in CRC.
In this study, we defined histologic grade according to the
conventional criteria, that is, tumors with greater than 50%
showing solid growth or lacking gland formation were
considered high grade. This was applied to medullary
2435
carcinomas as well, a type of CRC first described by
Jessurun et al [16] and incorporated by the World Health
Organization classification. We observed that a significant
subset (43%) of MMR-abnormal/ARID1A-abnormal tumors
was of medullary type, with greater than 50% of the tumor
having medullary histology, whereas only 11% of the MMR-
abnormal/ARID1A-normal tumors were of medullary type.
In general, although medullary carcinomas are morpholog-
ically similar to poorly differentiated/undifferentiated ade-
nocarcinoma, they are associated with lymphocytic
infiltration and frequently associated with MSI [17,18].
Clinically, both medullary carcinoma and MSI carcinomas of
other histologic subtypes are believed to have a better
prognosis [16,19,20] and be less likely to present with nodal
involvement or advanced N or M stage [21]. For this reason,
the World Health Organization recommends that all MSI
CRCs be regarded as low grade, irrespective of their
histologic grade [22]. On the other hand, however, there
are literature data showing that (1) there exist certain
medullary carcinomas that fare worse than their poorly
differentiated counterparts [23], and (2) some MSI cancers
behave more aggressively than others [24]. In this context,
our finding of the MSI cancers that were ARID1A abnormal
showing overrepresentation of medullary histology yet
unexpected high TNM staging may serve to indicate that
ARID1A abnormality may be linked to a specific subset of
sporadic MSI cancers that is biologically more virulent. As
such, ARID1A loss may bear prognostic significance.
Notably, ARID1A loss has been associated with tumor
grade and stage in other organs as well. In a study of ovarian
clear cell carcinoma, Katagiri et al [25] reported an
association between ARID1A loss and advanced Interna-
tional Federation of Gynecology and Obstetrics (FIGO) stage
and a shorter progression-free survival. Furthermore, their
study suggested that tumors with loss of ARID1A were more
likely to be resistant to platinum-based chemotherapy.
Similar adverse prognostic implication of ARID1A loss
has also been observed in endometrial [26-28] and gastric
[29] cancers. Notably, however, inconsistent reports also
exist. For example, in an analysis of 111 uterine endome-
trioid carcinomas, Rahman et al [30] failed to detect any
significant association between ARID1A loss and the
common clinicopathological features including FIGO stage
and grade. In our study, we also failed to demonstrate a
statistically significant difference in the 5-year overall
survival between ARID1A-normal and ARID1A-abnormal
cases. The limited number of ARID1A-abnormal cases may
have limited our survival analysis. We did indeed observe a
trend toward abnormal ARID1A associating with a worse
outcome among the MMR-abnormal cases. In addition to the
limitations caused by limited number of cases, the discrepant
results among the literature studies may also be due to
differences in organ- and histology type–specific oncogenic
pathways. Attention toward such organ- and histology type–
specific differences is warranted, as efforts on understanding
the role of ARID1A in various tumors are ongoing,
2436
In conclusion, our study indicates that a small but
significant subset of CRCs shows abnormal ARID1A, and
most of these tumors are sporadic MSI cancers with MLH1
promoter hypermethylation and loss of both MLH1and PMS2.
When compared with ARID1A-normal MSI cancers,
ARID1A-abnormal MSI cancers tend to present at a more
advanced TNM stage, suggesting that ARID1A loss might
identify a specific subset of MSI CRCs that is biologically
more aggressive. These findings warrant further investigation.
Notably, it has been speculated that the role of ARID1A and
other related members of the SWI/SNF complex in tumors
may be exploited for the development of novel therapeutic
strategies [1]; as such strategies become available, detection of
the ARID1A protein by IHC may also find application in
predicting tumor response to target-selective agents.
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