Non-Hodgkinlymphomas: high expression of Ezh2 in the aggressive variants and dominance of polycomb repressive complex

PRC1.4 over PRC1.2

Virchows Archiv

Lamia Abd Al-Kader1,5, Takashi Oka1, Katsuyoshi Takata１,Xu Sun1,Hiaki Sato2, Ichiro Murakami1,3,Tomohiro Toji1, Akihiro Manabe1, Hiroshi Kimura4 and Tadashi Yoshino1

1Department of Pathology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan; 2Department of Medical Technology, Graduate School of Health Sciences,Okayama University, Okayama, Japan;3Department of Molecular Pathology, Tottori University Medical School; 4Nuclear Dynamics Group, GraduateSchool of Frontier Science, Osaka University; 5Departmentof Pathology, Faculty of Medicine, Mansoura University, Egypt.

Corresponding author: Takashi Oka, PhD., DMSc.

Address: Department of Pathology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-chou, Kita-ku, Okayama 700-8558, Japan

E-mail:

Phone: (+81) 86-235-7150, Fax: (+81) 86-235-7156

Running title: Ezh2 correlates with aggressiveness

Table S1. Patient age, Gender and tumor location of B- and T/NK-cell neoplasms tested

Malignant lymphoma / Gender / Age range (mean age) / Nodal/ Total
M / F / Specimen
HD / 8 / 2 / 38-92 (75) / 2/10
BL / 10 / 2 / 6-70 (40.09) / 2/12
DLBCL-GCB / 8 / 6 / 29-92 (62.93) / 4/14
DLBCL non GCB / 6 / 8 / 28-83 (68.79) / 2/14
FLG3 / 4 / 4 / 34-82 (62) / 7/8
MCL / 8 / 1 / 56-75 (68.13) / 7/9
FLG1- 2 / 12 / 7 / 40-80 (61.42) / 14/19
CLL/SLL / 5 / 0 / 66-78 (69) / 5/5
PCM / 4 / 3 / 56-80 (64.75) / 0/7
PC / 4 / 2 / 45-84 (61.83) / 1/6
LPL / 4 / 1 / 73-83 (79) / 0/5
MALT lymphoma / 3 / 7 / 28-84 (62.4) / 0/10
B-cell neoplasms / 76 / 33 / 6-92 (62.38) / 44/109
T-LBL / 8 / 4 / 12-77 (43.42) / 8/12
ATLL / 13 / 5 / 38-81 (60.06) / 5/18
ALCL / 5 / 4 / 20-80 54.44) / 6/9
PTCL / 8 / 12 / 42-90 (69.74) / 12/20
NK/T / 5 / 5 / 14-79 (52.30) / 1/10
AITL / 4 / 2 / 58-90 (71.20) / 6/6
CTCL / 2 / 1 / 35-55 (51) / 0/3
T- and NK-cell neoplasms / 45 / 33 / 12-90 (58.54) / 38/78

HD: Hodgkin lymphoma, BL: Burkitt lymphoma, DLBCL-GCB: Diffuse large cell B cell lymphoma germinal centre type, DLBCL non GCB: Diffuse large B- cell lymphoma non germinal centre type, FLG3:Follicular lymphoma grade3, FLG1-2: Follicular lymphoma grade 1-2, MCL: Mantle cell lymphoma, CLL/SLL: Chronic lymphocytic leukemia/small lymphocytic lymphoma, PCM: Plasma cell myeloma, PC: Plasmacytoma, LPL: Lymphoplasmacytic lymphoma, MALT: Mucosa associated lymphoid tissue, T-LBL: T-cell lymphoblastic lymphoma, ATLL: Adult T-cell leukemia/lymphoma, ALCL: Anaplastic large cell lymphoma, , PTCL: Peripheral T-cell lymphoma, NK/T: Natural killer/T-cell lymphoma, AITL: Angioimmunoblastic T-cell lymphoma, CTCL: Cutaneous T-cell lymphoma

Table S2.Culture condition and origin of cell lines

Cell origin / Cell line name / Culture condition
T-cell acute lymphoblastic leukemia (T-LBL)
HTLV-1-immortalized T-cell
ATLL patient derived non-leukemic cell
ATLL patient derived leukemic cell
Natural killer/T cell (NK/T)
Burkitt Lymphoma (BL)
B-cell acute lymphoblastic leukemia (B-LBL)
Diffuse large B- cell lymphoma (DLBCL)
Follicular lymphoma(FL)
EBV- infected cell to FL18
EBV-immortalized normal B-cell(LCL)
Plasma cell myeloma
Cervical cancer cell
Human mesenchymal stem cell / Jurkat
MOLT4
IWA1
MT4
MT2
ED50823
ATL16T
ATL45T
ED40515
ATL55T
NK-L
Ramos
Dawdi
Raji
Scott
DLBCL2
SU-DHL-4
FL218
FL318
FL18
FL18EB
TW-EBV-LCL
KMS-21
KMS-34
KMS-12-PE
HeLa
hMSCs / RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS, 20u/ml IL2
RPMI1640, 10%FCS
RPMI1640, 10%FCS, 20u/ml IL2
RPMI1640, 10%FCS
RPMI1640, 10%FCS, 20u/ml IL2
RPMI1640, 15%FCS,100u/ml IL2
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
RPMI1640, 10%FCS
DMEM, 5%FCS
DMEM, 5%FCS

HTLV-1: Human T- cell leukemia virus-1, EBV: Epstein Barr virus.

Table S3. Difference in expression of Bmi-1, Ezh2 and Ki67 among different subtypes of T-cell

non Hodgkin lymphoma

Marker / ATLL / PTCL / T-LBL / NKT / AITL / ALCL
Bmi-1 / PTCL / - 0.480
P=1.000
T-LBL / - 0.780
P=0.605 / - 0.300
P=1.000
NKT / - 0.200
P=1.000 / 0.280
P=1.000 / 0.580
P=1.000
AITL / - 0.700
P=1.000 / - 0.220
P=1.000 / 0.080
P=1.000 / - 0.500
P=1.000
ALCL / - 0.530
P=1.000 / - 0.050
P=1.000 / 0.250
P=1.000 / - 0.330
P=1.000 / 0.170
P=1.000
CTCL / -0.530
P=1.000 / -0.050
P=1.000 / 0.250
P=1.000 / - 0.330
P=1.000 / 0.170
P=1.000 / 0.000
P=1.000
Ezh2 / PTCL / 0.106
P=1.000
T-LBL / - 0.361
P=1.000 / - 0.467
P=1.000
NKT / 0.156
P=1.000 / 0.050
P=1.000 / 0.517
P=1.000
AITL / 0.222
P=1.000 / 0.117
P=1.000 / 0.5837
P=1.000 / 0.067
P=1.000
ALCL / 0.056
P=1.000 / - 0.050
P=1.000 / 0.417
P=1.000 / -0.1000
P=1.000 / -0.167
P=1.000
CTCL / - 0.444
P=1.000 / - 0.550
P=1.000 / -0.083
P=1.000 / -0.600
P=1.000 / -0.667
P=1.000 / - 0.500
P=1.000
Ki-67 / PTCL / 0.043
P=1.000
T-LBL / -0.063
P=1.000 / -0.105
P=1.000
NKT / 0.038
P=1.000 / -0.005
P=1.000 / 0.1000
P=1.000
AITL / -0.063
P=1.000 / -0.105
P=1.000 / 0.000
P=1.000 / - 0.1000
P=1.000
ALCL / -0.063
P=1.000 / -0.105
P=1.000 / 0.000
P=1.000 / - 0.100
P=1.000 / 0.000
P=1.000
CTCL / -0.063
P=1.000 / -0.105
P=1.000 / 0.000
P=1.000 / - 0.100
P=1.000 / 0.000
P=1.000 / 0.000
P=1.000

One way ANOVA with Bonferroni correction, upper panel is the mean difference (Significant P <0.05 level).ATLL: Adult T-cell leukemia/lymphoma, PTCL: Peripheral T-cell lymphoma, T-LBL: T-cell lymphoblastic Lymphoma, NK/T Natural killer/T-cell lymphoma,AITL: Angioimmunoblastic T-cell lymphoma, ALCL: Anaplastic large cell lymphoma, CTCL: Cutaneous T-cell lymphoma.