CI, Confidence Interval; HR, Hazard Ratio

Revised manuscript # ASO-2014-08-1562. Inamura K et al. Signet-ring cell/mucinous colorectal cancer. Supplementary Tables. Page 1

Supplementary Table 1. Studies on prognostic significance of signet-ring cell carcinoma or mucinous carcinoma in colorectal carcinoma

Ref. / Authors (Year) / Sample size / Tumor location / Disease stage / N of signet-ring cell carcinoma (N of events) / N of mucinous carcinoma
(N of events) / Multivariate HR (95% CI; signet-ring cell carcinoma vs. conventional adenocarcinoma as a referent) / Multivariate HR (95% CI; mucinous carcinoma vs. conventional adenocarcinoma as a referent) / Variables examined in multivariate analysis
2 / Kang et al.
(2005) / 164,628 / Colon and rectum / I-IV / 1,522 (-) / 16,991 (-) / 1.37 (1.27-1.48) / 1.01 (0.98-1.04) / Covariates were age, sex, race/ethnicity, marital status, tumor location, stage at presentation, tumor grade, receipt of cancer-directed surgery, and receipt of radiation.
3 / Chew et al.
(2010) / 2,764 / Colon and rectum / I-IV / 30 (-) / 167 (-) / 1.9 (1.1-3.0) / 1.0 (0.7-1.3) / Covariates were age, sex, ethnicity, tumor location, tumor grade, histological subtype, T-stage, N-stage, perineural invasion, vascular emboli, and stage of disease.
4 / Shin et al.
(2011) / 368 / Rectum / II-III / - / 23 (-) / - / 2.36 (1.05-5.3) / Covariates were age, sex, comorbidity, pathologic nodal involvement, and perineal resection.
5 / Hyngstrom et al.
(2012) / 244,794 / Colon and rectum / I-IV / colon:
1,812 (-);
rectum:
448 (-) / colon:
21,037 (-);
rectum:
4,509 (-) / colon:
1.42 (1.33-1.51);
rectum:
1.57 (1.38-1.77) / colon:
1.03 (1.00-1.06);
rectum:
1.22 (1.16-1.29) / Adjusted by tumor stage and year of diagnosis. Covariates were age, sex, race, grade, chemotherapy, radiotherapy, and cancer-directed surgery.
6 / Nitsche
et al.
(2013) / 3,479 / Colon and rectum / I-IV / 30 (-) / 375 (-) / 2.5 (1.6-3.8) / 1.0 (0.83-1.2) / Covariates were sex, tumor location, stage (T, N, M), lymphatic invasion, angioinvasion, tumor grade, surgical resection margin (R)
Inamura
et al.
(current study) / 1,336 / Colon and rectum / I-IV / 20 (13) / 140 (45) / 4.53 (2.53-8.12) / 0.82 (0.54-1.23) / Adjusted by stage. Covariates were sex, age, year of diagnosis, family history of colorectal carcinoma, tumor location, tumor differentiation, microsatellite instability, CpG island methylator phenotype, LINE-1 methylation, and KRAS, BRAF, and PIK3CA mutations

CI, confidence interval; HR, hazard ratio.

Revised manuscript # ASO-2014-08-1562. Inamura K et al. Signet-ring cell/mucinous colorectal cancer. Supplementary Tables. Page 1

Supplementary Table 2. Microsatellite instability (MSI) status in colorectal carcinoma and patient mortality, stratified by the proportion of

signet-ring cell component

Colorectal carcinoma-specific mortality / Overall mortality
N of cases / N of events / Univariate
HR (95% CI) / Stage-stratified HR (95% CI) / Multivariate HR (95% CI) / N of events / Univariate
HR (95% CI) / Stage-stratified HR (95% CI) / Multivariate HR (95% CI)
Signet-ring cell component=0%
MSS / 934 / 280 / 1 (referent) / 1 (referent) / 1 (referent) / 499 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 141 / 12 / 0.26
(0.14-0.46) / 0.34
(0.19-0.60) / 0.29
(0.16-0.53) / 67 / 0.83
(0.64-1.07) / 0.95
(0.73-1.23) / 0.79
(0.60-1.05)
Signet-ring cell component=1-50%
MSS / 69 / 37 / 1 (referent) / 1 (referent) / 1 (referent) / 45 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 44 / 8 / 0.27
(0.13-0.58) / 0.57
(0.26-1.25) / 0.45
(0.20-1.01) / 17 / 0.47
(0.27-0.83) / 0.78
(0.44-1.37) / 0.55
(0.31-0.99)
Signet-ring cell component>50%
MSS / 12 / 8 / 1 (referent) / 1 (referent) / 1 (referent) / 10 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 5 / 3 / 0.70
(0.18-2.67) / 1.21
(0.31-4.68) / 0.53
(0.13-2.16) / 4 / 0.56
(0.18-1.81) / 0.73
(0.23-2.38) / 0.37
(0.11-1.23)
Pinteraction* / 0.33 / 0.058 / 0.15 / 0.10 / 0.50 / 0.12

The multivariate Cox regression modelswere stage-stratified. In addition to microsatellite instability, covariates initially includedsex, age at diagnosis, year of diagnosis, family history of colorectal carcinoma, tumor location, CpG island methylator phenotype, LINE-1 methylation,and KRAS, BRAF, and PIK3CA mutations.

*Pinteraction values (two-sided) were calculated by using the Wald test on the cross product of signet-ring cell component (0%, 1-50%, 50%; ordinal) and MSI status(MSI-high vs. MSS) in a Cox model.

CI, confidence interval; HR, hazard ratio; MSI, microsatellite instability; MSS, microsatellite stable.

Supplementary Table 3. Microsatellite instability (MSI) status in colorectal carcinoma and patient mortality, stratified by the proportion of

mucinous component

Colorectal carcinoma-specific mortality / Overall mortality
N of cases / N of events / Univariate
HR (95% CI) / Stage-stratified HR (95% CI) / Multivariate HR (95% CI) / N of events / Univariate
HR (95% CI) / Stage-stratified HR (95% CI) / Multivariate HR (95% CI)
Mucinous component=0%
MSS / 670 / 202 / 1 (referent) / 1 (referent) / 1 (referent) / 357 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 66 / 8 / 0.38
(0.19-0.77) / 0.37
(0.18-0.76) / 0.32
(0.15-0.65) / 36 / 0.99
(0.71-1.40) / 1.00
(0.70-1.41) / 0.82
(0.57-1.18)
Mucinous component=1-50%
MSS / 264 / 86 / 1 (referent) / 1 (referent) / 1 (referent) / 145 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 76 / 10 / 0.37
(0.19-0.71) / 0.62
(0.32-1.20) / 0.55
(0.28-1.08) / 33 / 0.75
(0.51-1.09) / 0.95
(0.65-1.40) / 0.80
(0.54-1.19)
Mucinous component>50%
MSS / 81 / 37 / 1 (referent) / 1 (referent) / 1 (referent) / 52 / 1 (referent) / 1 (referent) / 1 (referent)
MSI-high / 48 / 5 / 0.18
(0.07-0.46) / 0.41
(0.16-1.05) / 0.27
(0.10-0.72) / 19 / 0.47
(0.28-0.80) / 0.74
(0.43-1.27) / 0.51
(0.29-0.90)
Pinteraction* / 0.26 / 0.79 / 0.95 / 0.024 / 0.33 / 0.12

The multivariate Cox regression modelswere stage-stratified. In addition to microsatellite instability, covariates initially includedsex, age at diagnosis, year of diagnosis, family history of colorectal carcinoma, tumor location, CpG island methylator phenotype, LINE-1 methylation,and KRAS, BRAF, and PIK3CA mutations.

*Pinteraction values (two-sided) were calculated by using the Wald test on the cross product of mucinous component (0%, 1-50%, 50%; ordinal)and MSI status(MSI-high vs. MSS) in a Cox model.

CI, confidence interval; HR, hazard ratio; MSI, microsatellite instability; MSS, microsatellite stable.

Supplementary Table 4. Prognostic associations of each molecular feature (MSI, CIMP, KRAS, BRAF, PIK3CA, and

LINE-1 methylation)in colorectal carcinoma patients

Molecular feature / Colorectal carcinoma-specific mortality
Multivariate stage-stratified HR (95% CI) / P
MSI-high (vs. MSS) / 0.34 (0.22-0.54) / <0.0001
CIMP-high (vs. CIMP-low/negative) / 0.82 (0.52-1.29) / 0.38
KRAS mutation (vs. wild type) / 1.09 (0.87-1.37) / 0.43
BRAF mutation (vs. wild type) / 1.65 (1.20-2.27) / 0.0021
PIK3CA mutation (vs. wild type) / 0.88 (0.65-1.19) / 0.42
LINE-1 methylation (for a 30% decrease) / 1.77 (1.25-2.50) / 0.0013

The multivariate stage-stratified Cox regression modelinitially includedMSI, CIMP,KRAS, BRAF, PIK3CA, LINE-1 methylation,sex, age at diagnosis, year of diagnosis, family history of colorectal carcinoma, tumor location, tumor differentiation, signet-ring cell component variable (0%, 1-50%, 50%; ordinal) and mucinous component variable (0%, 1-50%, 50%; ordinal).

CI, confidence interval; CIMP, CpG island methylator phenotype; HR, hazard ratio; LINE-1, long interspersed nucleotide element-1; MSI, microsatellite instability;MSS, microsatellite stable.