Article
microRNA Expression in Sentinel Nodes from Progressing Melanoma Patients
Identifies Networks Associated with Dysfunctional Immune Response
Viviana Vallacchi, Chiara Camisaschi, Matteo Dugo, Elisabetta Vergani, Paola Deho, Ambra Gualeni, Veronica Huber, Annunziata Gloghini, Andrea Maurichi,
Mario Santinami, Marialuisa Sensi, Chiara Castelli, Licia Rivoltini, Monica Rodolfo
Table S1. Clinicopathological data for the sentinel node biopsy (SNB) samples analyzed in the study
ID
SNB
Status a
LN7
LN12
LN1
LN11
LN10
LN17
LN22
LN8
LN3
LN6
LN9
LN13
LN14
LN15
LN19
LN49
LN24
LN23
LN4
LN2
neg
neg
neg
neg
neg
neg
neg
neg
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
Genes 2016, 7, 124
Patient
status at 5y
FU b
N
N
N
N
N
N
N
N
PN
PN
PN
PN
PN
PN
PN
PN
PP
PP
PP
PP
SNB Tumor Burden at
Histopathology c
Stage after
Surgery d
Primary Tumor
Classification
Primary Tumor
Localization e
Gender
Age
neg
neg
neg
neg
neg
neg
neg
neg
6, m, 1.8, ip
single cells, ip
3, M, 2.88, ip
3, m, sc + ip
2, m, ip
1, m, ip
1, m, sc
6, M, 1.8, ip
4, M, ip
2, m, sc
1, 2.1, ip
1, M, ip
IB
IIB
IIA
IB
IB
IB
IB
IB
IIIA
IIIA
IIIB
IIIA
IIIA
IIIA
IIIA
IIIB
IIIC
IIIB
IIIC
IIIC
T1b
T3b
T3a
T1b
T2b
T1b
T1b
T2a
T3a
T3a
T3b
T2b
T3a
T3b
T2a
T3a
T2b
T2b
T3b
T3b
L
L
L
L
F
T
F
L
F
F
L
T
L
F
F
F
T
F
F
L
F
F
F
M
F
M
F
M
F
F
F
M
F
F
F
F
F
F
F
M
61
30
41
57
51
41
62
52
69
55
52
46
74
38
51
70
58
76
73
68
Relapse
LR/MET f
MET
LR
LR
LR
Tested for g
A
A
A
A
A
A
A
A
A+B
A
A+B
A+B
A+B
A
A+B
A
A+B
A+B
A+B
A+B
www.mdpi.com/journal/genes
Genes 2016, 7, 124
LN21
LN16
LN18
LN5
LN53
LN54
LN65
LN66
LN50
LN51
LN52
LN37
LN39
LN44
LN47
LN64
LN41
LN42
LN43
2 of 3
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
pos
PP
PP
PP
PP
PN
PN
PN
PN
PN
PN
PN
PP
PP
PP
PP
PP
PP
PP
PP
1, M, ip
2, M, ip
4, M, ip
1, m, ip
3,m,0.36,sc + ip
3,m, ip
2, M, 3.5, ip
single cells, ip
3,m, ip
2, M, 3.5, ip
single cells, ip
5, M, 2.4, sc
3, M, 2.6, ip + ex
single cells, ip
1, M, ip
1, M, ip
2,M,0.8, ip,ex
single cells, ip
single cells, ip
IIIC
IIIC
IIIC
IIIC
IIIB
IIIA
IIIA
IIIA
IIIA
IIIB
IIIA
IIIB
IIIC
IIIB
IIIC
IIIC
IIIB
IIIB
IIIB
T3b
T3a
T3b
T2a
T3a
T3b
T2a
T3a
T3A
T3B
T2A
T4b
T4b
T1b
T4b
na
T4B
T4B
T1B
L
T
L
F
L
F
T
F
T
L
T
L
F
L
T
L
L
L
T
M
F
M
M
M
F
F
F
F
F
F
F
M
F
F
F
M
F
F
66
57
69
67
46
74
54
55
54
53
51
71
73
65
29
46
52
65
49
MET
MET
MET
MET
MET
MET
MET
MET
LR
LR
LR
LR
A+B
A+B
A+B
A+B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
a. status of sentinel nodes relative to melanoma infiltration, pos: positive for metastases, neg: negative for metastases; b. disease course in 5 years of follow-up, N: negative,
PN: not progressing, PP: progressive disease; c. hystopathological evaluation of tumor burden; number of metastatic foci; size of metastases, m: <1 mm, M: >1mm, larger
diameter of the larger focus in mm; localization of metastases, ip: intraparenchimal, sc: subcapsular, ex: extracapsular; d. pathologic stage after complete lymph node
dissection (CLND) in SNB-positive patients; e. L: leg, F: foot, T: trunk. All SNB samples were inguinal; f. LR: locoregional relapse, MET: metastatic relapse;
g. A: tested by microRNA (miR) arrays; B: tested by quantitative real-time PCR (qRT-PCR); na: not available.
Genes 2016, 7, 124
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Figure S1. Overexpression of miR-30c-5p, miR-23a-3p and miR-4299 after transfection with miR
mimics. qRT-PCR analysis was performed in CD4+CD25− cells 72 h after transfection with miRNA
mimics or a scrambled control. The relative quantification was determined using 2−∆∆Ct method, using
cells transfected with scrambled control as calibrator. U6 snRNA was used as the internal reference.
* p < 0.0001 by unpaired t-test.
© 2016 by the authors. Submitted for possible open access publication under the
terms and conditions of the Creative Commons Attribution (CC-BY) license
(http://creativecommons.org/licenses/by/4.0/).