Article Figures & Data
Figures
- Fig. 1.
Immunohistochemical analyses of CD44, CD24, and ALDH1 in breast cancer. A, immunohistochemical staining patterns in normal breast tissue. CD24 and CD44 displayed uniform luminal and basal cell-specific pattern, respectively, whereas ALDH1 expression varied depending on the area of the tissue. B, representative examples of immunohistochemical analyses of IDC and DCIS according to tumor subtype using the indicated antibodies. In luminal A subtype, tumor cells positive for CD44 and CD24 (apical membranous pattern) can be seen whereas ALDH1 was not detected. HER2+ tumor is negative for CD44, and is positive for CD24 (membrano-cytoplasmic pattern) and ALDH1. In basal-like subtype, tumor cells are positive for CD44 and ALDH1, but negative for CD24. C, representative examples of double IHC for CD44 (brown) and CD24 (red) in IDC and in DCIS.
- Fig. 2.
Immunohistochemical analyses of genes differentially expressed between CD44+ and CD24+ breast cancer cells. A, representative examples of immunohistochemical staining patterns observed in invasive breast tumors using antibodies against the indicated proteins. A subset of markers that were highly expressed in CD44+ compared with CD24+ breast cancer cells based on our prior study are expressed only in basal-like tumors (vimentin, osteonectin, and caveolin 1), whereas Connexin 43 is expressed both in luminal A and basal-like subtypes. Markers more abundantly expressed in CD24+ compared with CD44+ breast cancer cells (Keratin 18, MUC1, and GATA3) are expressed in luminal A tumors. B, representative examples of immunohistochemical staining patterns observed in DCIS using antibodies against the indicated proteins. Vimentin is expressed in basal-like but not in luminal A DCIS. Strong vimentin staining is detected in stromal cells in both tumor subtypes. In contrast, GATA3 is expressed in luminal A but not in basal-like DCIS. C, immunohistochemical analyses of the expression of markers between invasive (red asterisk) and in situ (black asterisk) components of the same tumor. CD44 is expressed in both invasive and in situ components of luminal A subtype. HER2+ tumors are negative for CD44; only stromal staining is detected. In basal-like IDC, keratin 18 is expressed only in the in situ component whereas vimentin is expressed in both invasive and in situ areas.
Tables
- Table 1.
Distributions of the tumors in the four histologic groups
Subtype Luminal A Luminal B HER2+ Basal-like Total IDC alone 35 75% 1 2% 5 11% 6 13% 47 IDC with DCIS 85 63% 16 12% 18 13% 16 12% 135 DCIS with microinvasion 11 31% 6 17% 14 40% 4 11% 35 Pure DCIS 41 71% 7 12% 9 16% 1 2% 58 Total 172 63% 30 11% 46 17% 27 10% 275 NOTE: Number of patients (and raw percentage) in each of the histologic groups who had each of these tumor subtypes.
- Table 2.
CD24 and CD44 expression pattern defined based on immunohistochemical analyses of adjacent sections for the two markers according to tumor subtype and histologic groups
Histologic groups Pattern Tumor subtypes Total Luminal A Luminal B HER2+ Basal-like P = 0.001 P = NS P = NS P = NS P = 0.0001 IDC alone* P = NS CD44 and CD24 7 (20) 0 (0) 0 (0) 0 (0) 7 (15) CD44 only 9 (26) 1 (100) 3 (60) 5 (83) 18 (38) CD24 only 4 (11) 0 (0) 1 (20) 0 (0) 5 (11) Lack of both 15 (43) 0 (0) 1 (20) 1 (17) 17 (36) Total 35 (100) 1 (100) 5 (100) 6 (100) 47 (100) IDC with DCIS P = NS CD44 and CD24 24 (28) 3 (19) 4 (22) 2 (13) 33 (24) CD44 only 30 (35) 6 (37) 2 (11) 10 (63) 48 (36) CD24 only 10 (12) 4 (25) 5 (28) 0 (0) 19 (14) Lack of both 21 (25) 3 (19) 7 (39) 4 (25) 35 (26) Total 85 (100) 16 (100) 18 (100) 16 (100) 135 (100) DCIS with microinvasion P = NSAA CD44 and CD24 4 (36) 3 (50) 3 (21) 0 (0) 10 (29) CD44 only 5 (46) 2 (33) 1 (7) 4 (100) 12 (34) CD24 only 2 (18) 1 (17) 7 (50) 0 (0) 10 (29) Lack of both 0 (0) 0 (0) 3 (21) 0 (0) 3 (9) Total 11 (100) 6 (100) 14 (100) 4 (100) 35 (100) Pure DCIS† P = NS CD44 and CD24 15 (37) 3 (43) 3 (33) 1 (100) 22 (38) CD44 only 21 (51) 3 (43) 4 (44) 0 (0) 28 (48) CD24 only 4 (10) 1 (14) 0 (0) 0 (0) 5 (9) Lack of both 1 (2) 0 (0) 2 (22) 0 (0) 3 (5) Total 41 (100) 7 (100) 9 (100) 1 (100) 58 (100) Total P = 0.0006 CD44 and CD24 50 (29) 9 (30) 10 (22) 3 (11) 72 (26) CD44 only 65 (38) 12 (40) 10 (22) 19 (70) 106 (39) CD24 only 20 (12) 6 (20) 13 (28) 0 (0) 39 (14) Lack of both 37 (22) 3 (10) 13 (28) 5 (19) 58 (21) Total 172 (100) 30 (100) 46 (100) 27 (100) 275 (100) NOTE: Numbers in parentheses indicate column percentage. P values were calculated using Fisher's exact tests.
Abbreviations: NS, not significant (crude, unadjusted P > 0.05); NSAA, not significant after adjustment (using Holm's method).
- Table 3.
Presence of CD44+/CD24− and CD44−/CD24+ cells defined by double IHC according to tumor subtypes and histologic groups
Cell staining Tumor subtypes Total CD44+/CD24 CD24− cells CD44−/CD24+ cells Luminal A Luminal B HER2+ Basal-like P < 0.0001 P = 0.0009 P = NS P = NS P = NS IDC alone* P = NS Positive Positive 1 (3) 0 (0) 0 (0) 1 (17) 2 (4) Positive Negative 19 (54) 1 (100) 3 (60) 5 (83) 28 (60) Negative Positive 6 (17) 0 (0) 1 (20) 0 (0) 7 (15) Negative Negative 9 (26) 0 (0) 1 (20) 0 (0) 10 (21) Total 35 (100) 1 (100) 5 (100) 6 (100) 47 (100) IDC with DCIS P < 0.0001 Positive Positive 10 (12) 2 (13) 2 (11) 1 (6) 15 (11) Positive Negative 43 (51) 8 (50) 5 (28) 15 (94) 71 (53) Negative Positive 16 (19) 6 (38) 8 (44) 0 (0) 30 (22) Negative Negative 16 (19) 0 (0) 3 (17) 0 (0) 19 (14) Total 85 (100) 16 (100) 18 (100) 16 (100) 135 (100) DCIS with microinvasion P = NS Positive Positive 6 (55) 3 (50) 3 (21) 1 (25) 13 (37) Positive Negative 4 (36) 2 (33) 4 (29) 3 (75) 13 (37) Negative Positive 1 (9) 1 (17) 7 (50) 0 (0) 9 (26) Negative Negative 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) Total 11 (100) 6 (100) 14 (100) 4 (100) 35 (100) Pure DCIS† P = NS Positive Positive 12 (29) 2 (29) 2 (22) 0 (0) 16 (28) Positive Negative 23 (56) 3 (43) 5 (56) 1 (100) 32 (55) Negative Positive 4 (10) 0 (0) 1 (11) 0 (0) 5 (9) Negative Negative 2 (5) 2 (29) 1 (11) 0 (0) 5 (9) Total 41 (100) 7 (100) 9 (100) 1 (100) 58 (100) Total P = 0.0006 Positive Positive 29 (17) 7 (23) 7 (15) 3 (11) 46 (17) Positive Negative 89 (52) 14 (47) 17 (37) 24 (89) 144 (52) Negative Positive 27 (16) 7 (23) 17 (37) 0 (0) 51 (19) Negative Negative 27 (16) 2 (7) 5 (11) 0 (0) 34 (12) Total 172 (100) 30 (100) 46 (100) 27 (100) 275 (100) - Table 4.
Comparison of the expression of 12 proteins between tumor subtypes in each of the four histologic groups
IDC IDC alone IDC with DCIS Luminal A (n = 35) Basal-like (n = 6) HER2+ (n = 5) P Luminal A (n = 85) Basal-like (n = 16) HER2+ (n = 18) Luminal B (n = 16) P CD44+ cell genes Vimentin 7 (20) 6 (100) 2 (40) 0.0003 10 (12) 13 (81) 2 (11) 1 (6) <0.0001 Osteonectin 0 (0) 3 (50) 1 (20) 0.001 6 (7) 4 (25) 0 (0) 1 (6) NS Connexin 43 16 (46) 2 (33) 1 (20) NS 35 (41) 7 (44) 0 (0) 1 (6) 0.0001 ALDH1 0 (0) 2 (33) 0 (0) NSAA 1 (1) 3 (19) 4 (22) 0 (0) 0.001 Caveolin1 1 (3) 3 (50) 0 (0) NSAA 0 (0) 2 (12) 0 (0) 0 (0) NSAA CD44 16 (46) 5 (83) 3 (60) NS 54 (64) 12 (75) 6 (33) 9 (56) NS CD44 median 14 (40) 5 (83) 2 (40) NS 44 (52) 11 (69) 5 (28) 6 (38) NS EPCR 1 (3) 0 (0) 1 (20) NS 2 (2) 1 (6) 0 (0) 1 (6) NS CD24+ cell genes High pos CK18 22 (63) 0 (0) 0 (0) 0.0002 49 (58) 1 (6) 9 (50) 6 (38) 0.0008 GATA3 29 (83) 0 (0) 1 (20) <0.0001 78 (92) 1 (6) 6 (33) 11 (69) <0.0001 GATA3 median 21 (60) 0 (0) 1 (20) 0.004 56 (66) 0 (0) 1 (6) 6 (38) <0.0001 High pos MUC1 23 (66) 0 (0) 0 (0) 0.0002 34 (40) 2 (13) 6 (33) 7 (44) NS Claudin7 9 (26) 2 (33) 0 (0) NS 28 (33) 2 (13) 5 (28) 6 (38) NS CD24 11 (31) 0 (0) 1 (20) NS 34 (40) 2 (13) 9 (50) 7 (44) NS DCIS DCIS with microinvasion Pure DCIS Luminal A (n = 11) Luminal B (n = 6) HER2+ (n = 14) Basal-like (n = 4) P Luminal A (n = 41) Luminal B (n = 7) HER2+ (n = 9) P CD44+ cell genes Vimentin 2 (18) 2 (33) 1 (7) 3 (75) NSAA 4 (10) 1 (14) 4 (44) NSAA Osteonectin 0 (0) 2 (33) 1 (7) 1 (25) NS 1 (2) 1 (14) 0 (0) NS Connexin 43 1 (9) 0 (0) 0 (0) 1 (25) NS 21 (51) 1 (14) 0 (0) 0.003 ALDH1 0 (0) 0 (0) 1 (7) 0 (0) NS 0 (0) 1 (14) 1 (11) NS CD44 9 (82) 5 (83) 4 (29) 4 (100) NSAA 36 (88) 6 (86) 7 (78) NS CD44 median 4 (36) 4 (67) 4 (29) 3 (75) NS 24 (59) 3 (43) 4 (44) NS EPCR 4 (36) 0 (0) 0 (0) 0 (0) NSAA 4 (10) 0 (0) 0 (0) NS CD24+ cell genes High pos CK18 4 (36) 3 (50) 11 (79) 0 (0) NSAA 32 (78) 5 (71) 4 (44) NS GATA3 9 (82) 3 (50) 5 (36) 0 (0) NSAA 38 (93) 5 (71) 0 (0) <0.0001 GATA3 median 6 (55) 0 (0) 0 (0) 0 (0) 0.0008 29 (71) 3 (43) 0 (0) 0.0001 High pos MUC1 7 (64) 3 (50) 5 (36) 0 (0) NS 24 (59) 3 (43) 4 (44) NS Claudin7 5 (45) 1 (17) 1 (7) 1 (25) NS 22 (54) 0 (0) 2 (22) NSAA CD24 6 (55) 4 (67) 10 (71) 0 (0) NS 19 (46) 4 (57) 3 (33) NS NOTE: Numbers in parentheses indicate percentage of patients who were coded as positive for the binary marker variables (column percentage). P values were calculated using Fisher's exact tests. Significance levels were adjusted using Holm's method. Luminal B subtype was omitted in IDC alone group because there was only one case in this group. Basal-like subtype was omitted in pure DCIS group because there was only one case in this group. All DCIS tumors were negative for Caveolin1.
Abbreviation: High pos, higly positive.
Supplementary Data
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Volume 16, Issue 3
