Use of Histone Deacetylase 8 (HDAC8), a New Marker
of Smooth Muscle Differentiation, in the Classification
of Mesenchymal Tumors of the Uterus
Laurence de Leval, MD, PhD,* David Waltregny, MD, PhD,w Jacques Boniver, MD, PhD,*
Robert H. Young, MD,z Vincent Castronovo, MD, PhD,w and Esther Oliva, MDz
Am J Surg Pathol Volume 30, Number 3, March 2006
ESTs vs. SMTs
ESTs may be hypocellular secondary to fibrous or myxoid background
smooth muscle tumors.
leiomyoma variants, typically HCL, or less frequently highly cellular intravenous leiomyomatosis
ESTs.
markers of smooth muscle
1) smooth muscle alpha-actin (SMA) actin isoform mostly restricted to smooth mucle cells, myoe
pithelial cells, and myofibroblasts
2) Desmin Intermediate filament also expressed in some ESTs.
markers of smooth muscle
3) h-caldesmon a cytoplasmic protein ,interactions with actin a
nd tropomyosin more specific than desmin in the diagnosis of
uterine SMTs.
4) smooth muscle myosin heavy chain widely used in the diagnosis of smooth muscle
tumors of the uterus.
marker of ESTs
CD10 specific marker of ESTs CD10 in a substantial proportion of SMTs, espe
cially leiomyosarcomas and highly cellular leiomyomas,
SMTs that more often may be mistaken for ESTs.
Histone deacetylases (HDACs)
Histone deacetylases (HDACs)
Histone core represses transcription in vivo and in vitro.
Deacetylzed histone amino termini decreasing their affinity for DNA
allow the termini to be displaced from the nucleosomeincreasing access to transcription factors
NATURE |VOL 389 | 25 SEPTEMBER 1997
Histone deacetylases 8(HDAC8)
mainly a cytosolic distribution expressed by cells showing smooth muscle differe
ntiation(visceral and vascular smooth muscle cells, myoepithelial cells , myofibroblasts)
not been assessed in mesenchymal neoplastic tissues, and it remains to be determined
can be used as a marker of smooth muscle differentiation in neoplastic conditions
Study Materials 15 typical leiomyomas (LMs), 9 highly cellular leiomyomas (HCLs), 8 epithelioid smooth muscle tumors (epithelioid SMT
s), 13 leiomyosarcomas (LMSs), 17 endometrial stromal tumors (ESTs) (4 endometrial stromal nodules and 13 low-grade en
dometrial stromal sarcomas) (3 with sex-cord-like differentiation and 5 with smoot
h muscle differentiation)
Study Materials
10 samples including myometrium and endometrium were examined as controls.
from the Pathology Departments of the Massachusetts General Hospital and of the C.H.U. Sart-Tilman, and from the consultation files of one of us (R.H.Y.).
Immunohistochemistry
All cases were stained with specific antibodies against HDAC8, desmin, h-caldesmon, a-SMA, smooth muscle myosin heavy chain, and CD10. using the avidin-biotin detection system
Negative controls were performed Cases were considered negative when less than 5
% of the tumor cells were immunoreactive.
Immunohistochemistry Five-micrometer, formalin-fixed, paraffin-embedded t
issue sections deparaffinized in xylene rehydrated in graded alcohols. blocking of the endogenous peroxidase activity with
0.3% hydrogen peroxide in methanol for 30 minutes heat-induced antigen retrieval cool down incubation with primary antibodies.
Result- Normal Uterine Tissues
HDAC8 strongly stained myometrial and vascular smooth muscle cells (Fig. 1A).
Result- Normal Uterine TissuesHDAC8 In the endometrium, endometrial stromal and glandular cells were uniformly negativearteriolar smooth muscle cells stained positive
Result- Normal Uterine Tissues
HDAC8 The same distribution of expression was observed in foci of adenomyosis
Result- Normal Uterine Tissues
HDAC8only stains the vascular walls in an endometrial polyp
Result- Normal Uterine Tissues
Desmin, h-caldesmon, and smooth muscle myosin showed the same staining pattern than that observed for HDAC8.
SMA displayed moderate but extensive staining of the endometrial stroma.
CD10 stained strongly and diffusely the endometrial stroma, and it was negative in the myometrium.
Result-Conventional Leiomyomas
HDAC8Conventional leiomyoma
moderate to strong HDAC8 expression in most tumor cells (90%–100%)
Result-Conventional Leiomyomas
smooth muscle myosin
conventional leiomyoma
strongly positive
Result-Conventional Leiomyomas
Desmin, h-caldesmon, and SMA were also diffusely and strongly expressed by all tumors.
CD10 showed variable positivity in six cases
(weak to moderate [2] and moderate to strong [4] in up to 60% of tumor cells).
Result-Highly Cellular LeiomyomasHDAC8
8 of 9 HCLs were positive
(ranging from 40% to 90% positive tumor cells)
(intensity of staining being weak in 1 case, moderate in 4 cases, and strong in 4 cases )
Result-Highly Cellular Leiomyomas
smooth muscle myosin
9/9 strongly expressed
(in 7/9 neoplasms at least 80% of the tumor cells.)
desmin –9/9 strongest positivity (3+ in all cases).
h-caldesmon – 9/9 varied from 10% to 90%.
Result-Epithelioid Smooth Muscle Tumors
HDAC8 (8/8)
diffuse cytoplasmic and membranous staining
weak (4 cases) moderate (3 cases)moderate to strong(1) positive cells varied from 40% to 100%
Result-Epithelioid Smooth Muscle Tumors
smooth muscle myosin(3/8)
diffuse cytoplasmic and membranous staining
(20% to 70% of neoplastic cells)
h-caldesmon – 1/8 in one third of the tumor cells.
Result-LeiomyosarcomasHDAC8 11/13Strongly positive (3) 15% - 90% cellsModerate positive (6) in 20% - 100% of cellsweak to moderate positivity (2) in 35% - 50% of cells
Result-LeiomyosarcomasSmooth muscle myosin (10/13)
strongly positive(9) in 25 - 100%
moderate staining (1) of 10% tumor cells
h-caldesmon –12/13 strongly positive (11) 15% to 100% cells weak staining (1) in 45% of cells
SMA –12/13 20% to 100% of tumor cells
CD10 –5/13 >30% of tumor cells, strong immunoreactivity (4)
Result-Endometrial Stromal Tumors
HDAC8 (0/9) negative in conventional ESTs
Result-Endometrial Stromal TumorsSmooth muscle myosin (0/9) negative in a conventional EST
Result-Endometrial Stromal Tumors
HDAC8 (0/3)negative in 3 ESTs with sex-cord differentiation
Result-Endometrial Stromal TumorsEST with smooth muscle differentiation
Result-Endometrial Stromal TumorsHDAC8 (5/5) EST with smooth muscle differentiationweak to moderate positity
staining restricted to areas of smooth muscle differentiation
Result-Endometrial Stromal TumorsSmooth muscle myosin(3/5)positive in an area of smooth muscle differentiation
*Only in areas of smooth muscle differentiation.
In conventional areas of EST (1) and in areas of smooth muscle differentiation (3).
In conventional areas of EST and in areas of smooth muscle differentiation (4) and only in metaplastic areas (1).
Discussion
☆Our current study confirms the lack of specificity of CD10
☆HDAC8 in LMSs tended to be less intense ☆ESTs may be positive for actin and desmin
☆SMA also stained endometrial stromal cells surrounding endometrial glands
Discussion-about CD10
CD10 Chu and Arber first reported
16/16 ESSs versus 2/19 SMTs (2 cellular leiomyomas)
several follow-up studies CD10 is expressed to variable extent in uterine SMTs, bein
g more commonly positive in LMSs and also other sarcomas
McCluggage WG, Sumathi VP, Maxwell P.Oliva E, Young RH, Amin MB, et al.Toki T, Shimizu M, Takagi Y, et al.
☆ desmin displayed the highest intensity and extent of expression In HCLs.
☆ all uterine epithelioid SMTs were HDAC8 positive, although the degree of positivity was usually weak to moderate.
☆most epithelioid SMTs were completely negative for both desmin and h-caldesmon
Discussion
HDAC8 expression is seen in the myometrium and vascular wall
s not in the endometrial stroma, endometrial glands, or en
dothelium current findings confirm the results of the original
work. Waltregny D, Glenisson W, Tran SL, et al. Histone deacetylas
e HDAC8 associates with smooth muscle alpha-actin and is essential for smooth muscle cell contractility. FASEB J. 2005;19:966–968.
☆HDAC8 and h-caldesmon only stained areas of smooth muscle differentiation, but HDAC8 was more sensitive in detecting smooth muscle differentiation
北醫 萬方醫院
HCL 27 Pt’(95~22y/o)
1 Pt’
LMS 6 Pt’ (78~30 y/o)
2 Pt’ (44~57y/o)
Epithelioid SMT
2 Pt’ (45~53 y/o)
1 Pt’ (36 y/o)
EST 8 Pt’ (36~51 y/o)
6 Pt’ (42~55 y/o)