演講人：陳玉昆副教授 高雄醫學大學附設醫院口腔病理科主任 暨主治醫師 日 期： 2010 年 12 月 24 日地 點：第一教學大樓 B1 演藝廳

Part I ： Case report-Mandibular aneurysmal bone cystPart II ： Case report-Pleomorphic adenoma with extensive necrosis Part II ： Experimental study-Ameloblastoma

Content

Part I Chin Dent J 1990;19:33-41

Aneurysmal bone cyst of the mandible – a case report

Yuk-Kwan Chen, Li-Min Lin, Cheng-Chung Lin and Steven Lai

Chin Dent J 1990;19:33-41

Aneurysmal bone cyst of the mandible – a case report

Yuk-Kwan Chen, Li-Min Lin, Cheng-Chung Lin and Steven Lai

IHC: Factor VIII (-ve staining)

+internalcontrol

Citation: 3 times

Oral Diseases 2004;10:54-9

Part II

Expression of osteonectin/secreted protein acidic and rich in cysteine (SPARC) and matrix metalloproteinases (MMPs) in ameloblastoma

Shen, Ling-ChangChen, Yuk-KwanShaw, Shyh-Yu

Part III

Ameloblastoma/ 釉質母細胞瘤• Odontogenic epithelial origin

• The most common clinically significant odontogenic tumor (59.3%~63.2%)

• A locally aggressive neoplasm

• Surgery

• High recurrent rate (50-90%)

Contemporary oral and maxillofacial pathology 1997, 126-34Contemporary oral and maxillofacial pathology 1997, 126-34

SPARC

• Secreted Protein Acidic and Rich in Cysteine• Osteonectin/BM-40/43-kd protein• Bone-specific protein linking mineral to collagen

• Multifunctional glycoprotein (tissue differentiation, embryo

development) • Modulate cell-matrix interactions and cell function

• Re-appear in response to tissue injury, inflammation, and remodeling, as well as in tumor growth

and metastasis

Termine et al. Cell 1981;26:99-105, FASEB J 8:163-73, 1994, Matrix biology 2000;19:816-827Termine et al. Cell 1981;26:99-105, FASEB J 8:163-73, 1994, Matrix biology 2000;19:816-827

Tissue specific of SPARC in tumorsTumor enhancer Description

Glioma Enhances invasion (migration)

Melanoma Promotes metastasis (proliferation and migration)

Non small cell lung cancer Favors invasion and migration

Invasive meningiomas Diagnostic marker (negative in benign/non-invasive type)

Tumor suppressor Description

Ovary carcinoma Induces apoptosis

Pancreatic carcinoma Target for aberrant methylation

Breast cancer cells Inhibits proliferation (MDA-MB-231)

Neuroblastoma Inhibits angiogenesis

Role is not the same in different tumors

Int J Cancer: 2006;118,310-316. Journal of cellular biochemistry 2004;92:679-90Int J Cancer: 2006;118,310-316. Journal of cellular biochemistry 2004;92:679-90

Colorectal carcinoma Tumor-associated blood vessels

Matrix metalloproteinases (MMPs)• Six subfamilies • Zinc- and calcium-dependent enzymes that degrade native

and denatured collagen and other extracellular components <Extracellular matrix (ECM) turnover>• Rise rapidly when tissues undergo remodeling, such as in

inflammation, wound healing, and cancer

• Gelatinase, MMP-2 and -9, are essential proteases for basement membrane-invasive events (nonfibrillar collagen type IV)

Circ Res 2003; 92:827-39, modified Circ Res 2003; 92:827-39, modified

Invasion of the surrounding tissue

Gene Therapy 2006;13:464–77Gene Therapy 2006;13:464–77

MMPs

Angiogenesis switch

Matrix metalloproteinases (MMPs) in the invasiveness of ameloblastoma

Histopathology 2004;45:65–72Histopathology 2004;45:65–72

Bone Neoplasm

Bone

Bone Bone

Neoplasm

Neoplasm Neoplasm

(a)Mitogens

(b)Mitogens

MMPs

(1,2,9)(1,2,9)MMPs

(c)MMPs inducebone resorption andrelease of mitogensfrom the bone matrix

(d)Mitogens induce proliferationof ameloblastoma cellsas cords branching outfrom the central portionsof the neoplasm

SPARC and MMPs

• SPARC expression in human melanoma cells directly correlated with MMP-2 and MMP-9 levels and activity

• SPARC induces MMP-2 activation in human breast cancer cell line

• Both SPARC and MMP-9 have impact on ECM deposition and angiogenesis in the tumor microenvironment

Nature medicine 1997;3,171-5, , Cancer research 1998;58,5529-36,Experimental biology and medicine online April 29,2008

Nature medicine 1997;3,171-5, , Cancer research 1998;58,5529-36,Experimental biology and medicine online April 29,2008

AIMS

• To verify the pathologic expression of SPARC in ameloblastoma

• To establish the association between the presence of SPARC and MMPs in a cohort of ameloblastoma patients.

IHC expression of MMP-1, -2, -9 and SPARC

MMP-1MMP-1 MMP-2MMP-2

MMP-9MMP-9 SPARCSPARC

Immunohistochemical reactivity

• Immunoreactivity for SPARC, MMP-1, MMP-2 & MMP-9 was evaluated and classified into four groups:

score 0-2 (+)

score 3-5 (++)

score 6-8 (+++)

score 9-12 (++++)

• Pearson’s χ2-test. P -values less than 0.05 were considered to indicate statistical significance

Am J Pathol 1996;149,:1449-57, modifiedAm J Pathol 1996;149,:1449-57, modified

Case no. SPARC MMP-1 MMP-2 MMP-9

1* ++ ++ ++ +++

2 +++ ++ ++++ ++++

3 + ++ + ++

4* ++++ ++ ++ ++++

5* +++ ++ ++++ ++++

6 +++ ++ + +++

7* +++ ++ ++ ++++

8 +++ +++ +++ ++++

9 +++ ++ ++ ++++

10 ++ +++ ++ +++

11 ++++ +++ ++++ ++++

12 ++++ + + ++++

Case no. SPARC MMP-1 MMP-2 MMP-9

13 ++++ + ++ ++++

14 +++ + ++ ++++

15* ++ ++ ++ ++

16 +++ ++ + ++++

17 +++ + +++ ++++

18 ++++ +++ + ++++

19* ++++ + + ++++

20* +++ + ++ ++++

21 ++++ ++ +++ ++++

22 ++++ ++ ++ ++++

23 ++++ ++ ++ ++++

Immunohistochemical reactivity

1. 在 ameloblastoma tissue 當中，的確有 SPARC 的不正常表現2. 不同的 sample 之間，同一個蛋白質，有不同的表現強度；同一個 sample 的不同蛋白質，也有不同表現強度

SPARC MMP-1 MMP-2 MMP-9

SPARC 0.8122 0.5984 0.0003*

MMP-1 0.8122 0.8374 0.5062

MMP-2 0.5984 0.8374 0.8644

MMP-9 0.0003* 0.5062 0.8644

Associations between the immunoscores of SPARC, MMP-1, -2 and -9

IHC of MMP-9 and SPARC

MMP-9MMP-9 SPARCSPARC

Co-expression of SPARC and MMP-9 ( Double stain IHC)

MMP-9MMP-9 SPARCSPARC

Co- expressionCo- expression

A

B C

Conclusion I

• First to identify the pathologic expression of SPARC in ameloblastoma tissues

• SPARC was elevated consistently in matched MMP-9 expression

SPARC protein

MMP-9

VEGF

Angiogenesis

Expression in

Ameloblastoma

Tumor Invasion

Pinheiro et al, 2004

Kumamoto et al, 2002

Hei

ssig

et a

l, 20

02

Framson & Sage, 2004

Kato et al, 2001

J Oral Pathol Med 2010;39:242-9 J Oral Pathol Med 2010;39:242-9

Conclusion II :Putative association between SPARC and MMP-9 in ameloblastoma

Thank you very much