Supplementary data 3:
Correlation of artemin expression with cancer type and clinical outcome
To determine the clinical relevance of artemin expression in human mammary carcinoma, we used the Oncomine cancer microarray database to analyse the expression profile of artemin in a variety of human cancers (Table S4). We compared the expression level of artemin in normal and cancer tissues for all the microarray data in the database.
Increased expression of artemin was observed in a number of common cancers, including lung, ovarian, cervix, pancreatic, skin, head and neck, seminoma and leukemia (Table S4).
Next, we analysed differential artemin expression patterns in breast cancer (Table S5).
Although artemin expression did not differ significantly between normal tissue and breast cancer, one study (Su et al.
, 2001) found significantly higher expression in breast cancer compared with other types of cancers ( p <0.001). Interestingly, artemin expression was significantly increased from atypical ductal hyperplasia to developed cancer ( p <0.042).
Within breast cancer, higher expression was observed in basal tumors compared to apocrine and luminal tumors (Farmer et al.
, 2005). Invasive ductal carcinoma was observed to have higher artemin expression compared to ductal carcinoma in situ (Radvanyi et al.
, 2005).
Elevated artemin expression was observed from grade 1 to 3 with a correlation factor of 0.274
(Ginestier et al.
, 2006). In addition, artemin expression was positively correlated with ER and progesterone receptor (PR) status. Three studies demonstrated that ER+ tumors had significantly higher artemin expression than did ER- tumors (Hess et al.
, 2006; Richardson et al.
, 2006; Bild et al.
, 2006). Significantly higher expression was found in PR+ tumors in two studies (Ma et al.
, 2004; Bild et al.
, 2006). Increased expression was also observed in male breast cancer patients compared with female patients (Bittner, 2005). HER2/neu (c-erbB-2)-
S3-1

positive tumors exhibited lower artemin expression than did HER2/neu-negative tumors
(Hess et al.
, 2006; Richardson et al.
, 2006).
Increased artemin expression was observed to predict breast cancer recurrence and poor outcome (Table S5). One study demonstrated that patients with recurrence of ER+ tumors exhibited higher expression than did those with 5-year disease-free survival (van de
Vijver et al.
, 2002). Two studies demonstrated that patients with poor outcome (death) exhibited higher expression of artemin than did those with 5-year disease-free survival
(Ivshina et al.
, 2006; Desmedt et al.
, 2007). Patients with distant metastasis had significantly higher artemin expression than did those with no distant metastasis (Desmedt et al.
, 2007).
Patients who exhibited a complete response to chemotherapy had significantly lower artemin expression than those who did not, with p <0.001 for the ER+ group and p <0.05 for all patients (Hess et al.
, 2006; Desmedt et al.
, 2007). Expression of artemin in mammary carcinoma is therefore significantly correlated with clinical outcome.
The expression profile of GDNF in a variety of human cancers and human breast cancers and normal tissues was also analyses with the Oncomine cancer microarray database
(Table S6). GDNF lacks the multiple highly significant correlations with disease and outcome observed for artemin.
S3-2

Table S4: Increased expression of artemin in human cancers compared with normal tissues. Data were taken from the cancer microarray database Oncomine
(www.oncomine.org).
Type of
Cancers
Lung
Tissue
Ovary
Cervix
Pancreas
Head &
Neck
Normal lung
Carcinoid
Normal lung
Lung adenocarcinoma
Normal lung
Small cell lung cancer
Normal lung
Squamous cell lung carcinoma
Normal lung
Squamous cell carcinoma
Normal lung
Adenocarcinoma
Normal ovary
Ovarian mucinous adenocarcinoma
Normal ovary
Ovarian clear cell adenocarcinoma
Normal ovary
Ovarian serous adenocarcinoma
Normal ovary
Ovarian endometrioid adenocarcinoma
Normal
Cervical Cancer
Microdissected normal pancreatic duct
Microdissected pancreatic tumor epithelia
Normal oral mucosa
Head & neck squamous cell carcinoma
Normal
Leukemia
Head And Neck Cancer
Normal bone marrow
Acute myeloid leukemia
Normal CD3+ Purified Peripheral Blood Cells
inv(14)(q11q32)-Positive T-Cell
Prolymphocytic Leukemia
Seminoma Normal testis
Adult male germ cell tumor
Bladder Normal bladder
Bladder carcinoma
20
11
14
13
41
14
41
4
37
8
42
6
23
8
13
4
8
4
5
19
20
4
Sample number
17
20
17
139
17
6
17
21
5
5
6
91
48
109
Median p-value References
-0.04
-0.69
0.01
-0.11
0.10
-0.47
-0.2
-1.24
-1.04
-0.26
1.18
0.57
0.69
-0.69
-0.12
-0.69
0.00
-0.69
0.07
-0.86
0.96
-0.75
-0.07
0.80
1.26
0.01
0.40
0.01
0.52
0.01
0.26
0.83
-0.72
-0.60
-0.19
0.03
6E-6
6.4E-5
9.9E-5
0.009
0.004
0.024
6.2E-6
8.4E-6
1E-5
1E-5
2E-4
0.024
0.001
9.1E-4
6E-6
2E-4
2.2E-5
7.6E-6
(Bhattacharj ee et al.
,
2001)
(Wachi et al.
, 2005)
(Stearman et al.
, 2005)
(Hendrix et al.
, 2006)
(Pyeon et al.
,
2007)
(Grutzmann et al.
, 2004)
(Ginos et al.
,
2004)
(Pyeon et al.
,
2007)
(Andersson et al.
, 2007)
(Durig et al.
,
2007)
(Korkola et al.
, 2006)
(Sanchez-
Carbayo et al.
, 2006)
Skin
Normal bladder - biopsy
Normal bladder mucosa - cystectomy
Superficial transitional cell carcinoma
Invasive transitional cell carcinoma
Normal skin
Benign nevus
Melanoma
9
5
27
13
7
18
45
-0.78
-0.73
-0.45
-0.69
-0.21
-0.35
-0.06
0.003
2.9E-4
(Dyrskjot et al.
, 2004)
(Talantov et al.
, 2005)
S3-3

Table S5. Expression of artemin in human normal mammary tissue and mammary carcinoma. Data were taken from the cancer microarray database Oncomine
(www.oncomine.org).
Analysis type
Analysis
Class (sample number)
3
Correlation
(up/down)
p-value References
Cancer vs. cancer
Tissue – Type
Breast Carcinoma -
Type
Breast Carcinoma -
Type
1
Various carcinoma tissues
¶
Apocrine
Tumor (6),
Luminal Tumor
(27)
Ductal
Carcinoma In
Situ (3)
2
Breast Ductal
Adenocarcinoma
(26)
Basal Tumor
(16)
Invasive Ductal
Carcinoma (33)

*


9.50E-07
0.017
0.042
(Su et al.
,
2001)
(Farmer et al.
, 2005)
(Radvanyi et al.
, 2005)
ER+ Breast
Carcinoma - Disease
Free Survival - 5 Years
No Disease
(164)
Relapse (51)

0.021
(van de
Vijver et al.
,
2002)
Prognosis
Breast Carcinoma -
Disease-Free Survival
- 5 Years
Alive (158) Dead (69)

0.028
(Ivshina et al.
, 2006)
Alive (135) Dead (56)

0.047
(Desmedt et al.
, 2007)
Tumor grade
Breast Carcinoma -
Distant Metastasis At
5 Years
Breast Carcinoma -
Grade
Breast Carcinoma -
Nottingham Grade
Breast Carcinoma -
ER Status
Breast Carcinoma - PR
Status
No Distant
Metastasis
(154)
Grade 1 (4)
1 (3)
Negative (51)
Negative (24)
Negative (48)
Negative (57)
Negative (6)
Distant
Metastasis (35)
Grade 2 (12)
2 (5)
Positive (82)
Positive (15)
Positive (110)
Positive (101)
Positive (34)
Grade
3 (39)
3 (2)

0.274
-0.657





0.048
0.043
0.039
0.024
0.035
4.7E-5
8.7E-4
0.029
Breast Carcinoma -
Sex
Female (128) Male (4)

7.60E-04
Misc.
Breast Atypical Ductal
Hyperplasia -
Developed Cancer
ER+ Breast
Carcinoma -
Chemotherapy
Response
ER+/- Breast
Carcinoma -
Chemotherapy
Response
Negative (4)
Residual
Disease (75)
Residual
Disease (99)
Positive (4)
Complete
Response (7)
Complete
Response (34)


*

0.042
2.90E-05
0.038
(Poola et al.
,
2005)
(Hess et al.
,
2006)
(Hess et al.
,
2006)
ER+ Breast
Carcinoma -
HER2/neu Status
Breast Carcinoma -
HER2/neu Status
Negative (67)
Negative (29)
Positive (15)
Positive (8)


0.03
0.042
(Hess et al.
,
2006)
(Richardson et al.
, 2006)
Breast Carcinoma -
Negative (69) Positive (6)

0.003
(Desmedt et
Angioinvasion al.
, 2007)
¶
Bladder/Ureter Carcinoma (8), Colorectal Adenocarcinoma (23), Gastroesophageal Adenocarcinoma (12),
Hepatocellular Carcinoma (7), Lung Adenocarcinoma (14), Lung Carcinoma - Squamous Cell (14), Ovarian
Adenocarcinoma - Serous Papillary (27), Pancreatic Adenocarcinoma (6), Prostate Adenocarcinoma (26),
Renal Cell Carcinoma - Clear Cell (11)
*

and

represent a increased and decreased expression in class 2 compared with class 1, respectively.
(Desmedt et al.
, 2007)
(Ginestier et al.
, 2006)
(Turashvili et al.
, 2007)
(Hess et al.
,
2006)
(Richardson et al.
, 2006)
(Bild et al.
,
2006)
(Bild et al.
,
2006)
(Ma et al.
,
2004)
(Bittner,
2005)
S3-4

Table S6: Expression of GDNF in human normal mammary tissue and mammary carcinoma. Data were taken from the cancer microarray database Oncomine (www.oncomine.org).
Analysis type* Analysis
Cancer vs.
Normal (3)
Cancer vs.
Cancer (12)
Tumour Grade
(11)
Prognosis
(all of 19)
Breast - Type
Breast Carcinoma - Type
Breast Carcinoma - Grade
Breast Carcinoma - Distant
Metastasis At 5 Years
Breast Carcinoma - Metastases - 5
Years
Breast Carcinoma - Bone
Metastasis-Free Survival - 5 Years
Breast Carcinoma - Lung
Metastasis Event-Free Survival - 5
ER+ Breast Carcinoma - Disease
Free Survival - 5 Years
ER- Breast Carcinoma - Disease
Free Survival - 5 Years
Breast Carcinoma - Disease-Free
Survival - 5 Years
Breast Carcinoma - Survival - 5
Years
Breast Carcinoma - Recurrence -
5 Years
Breast Carcinoma - Relapse - 5
Years
Breast Carcinoma - Overall
Survival - 5 Years
1
Class (sample number)
2
Normal Breast (7)
Breast
Carcinoma (40)
Apocrine Tumour (6),
Luminal Tumour (27)
Basal Tumour
(16)
1 (4)
No Distant Metastasis
(154)
Negative (51)
No Metastasis (49)
No Metastasis (51)
No Disease (164)
No Disease (135)
No Disease (45)
No Disease (32)
Alive (135)
Alive (158)
No Disease (196)
No Disease (180)
No Recurrence (26)
Alive (232)
Alive (121)
2 (12)
Distant
Metastasis (35)
Positive (46)
Metastasis (11)
Metastasis (14)
Relapse (51)
Relapse (66)
Relapse (27)
Relapse (28)
Dead (56)
Dead (69)
Relapse (79)
Relapse (93)
Recurrence (14)
Dead (48)
Dead (38)
No Recurrence (196) Recurrence (79)
Negative (42)
No Relapse (85)
Alive (166)
Positive (18)
Relapse (28)
Dead (8)
3
3 (39)
Correlation
(up/down)

**
↓
0.344
pvalue
0.047
0.022
0.01
0.174
0.875
0.396
0.735
0.192
0.321
0.665
0.499
0.298
0.134
0.35
0.455
0.179
0.405
0.855
0.35
0.666
0.918
0.353
References
(Richardson et al.
, 2006)
(Farmer et al.
, 2005)
(Ginestier et al.
, 2006)
(Desmedt et al.
, 2007)
(van't Veer et al.
, 2002)
(Minn et al.
,
2005)
(Minn et al.
,
2005)
(van de
Vijver et al.
,
2002)
(Wang et al.
,
2005)
(Wang et al.
,
2005)
(van de
Vijver et al.
,
2002)
(Desmedt et al.
, 2007)
(Ivshina et al.
, 2006)
(van de
Vijver et al.
,
2002)
(Wang et al.
,
2005)
(Ma et al.
,
2004)
(van de
Vijver et al.
,
2002)
(Pawitan et al.
, 2005)
(van de
Vijver et al.
,
2002)
(Ma et al.
,
2004)
(Sotiriou et al.
, 2006)
(Desmedt et al.
, 2007)
Misc. (51)
Breast Carcinoma - Oestrogen
Receptor Status
Breast Carcinoma - HER2/neu
Status
Negative (34)
Negative (52)
Positive (85)
Positive (3)
↓
↓
0.001
0.014
(Sotiriou et al.
, 2006)
(Ma et al.
,
2004)
Molecular Breast Carcinoma - p53 Mutation (Miller et al.
,
Wild Type (179) Mutant (72) ↓ 0.011
Alteration (9) Status 2005)
* Total number of studies is indicated in brackets. Only studies with a p value < 0.05 in T-test are shown except prognosis for which all studies are shown.
**

and

represent an increased and decreased expression in class 2 compared with class 1, respectively.
S3-5

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