Georgian Scientists/ 4(02), 2022 117 Georgian Scientists Vol. 4 Issue 2, 2022 https://doi.org/10.52340/gs.2022.04.02.09 , Luminal A Luminal B 1; 2; 3 1 , 2 3 , (FFPE) (NST) 55 . : ; ; E - ; ; Her2, Ki67; ,, ’’ H&E Qupath- ( 0.3.2). Luminal B . , . , , . : ; ; (TB); Georgian Scientists/ 4(02), 2022 118 [1]. [2]. , , . EMT : 1 , ; 2 - ; 3 , [1,2,3]. EMT ; E- N- , . EMT , [4] . , . , (MET). MET , [2]. EMT- TGF- , Notch Wnt ; , (miRNAs) [5,6]. , (ER) [9]. - : Luminal A, Luminal B; Her2 ; Georgian Scientists/ 4(02), 2022 119 , . , KRAS , KRAS , SNAI2- [4]. [4]. (Tumor budding) [2]–[4]. , , (TB) , . ( - ) ( ). , , , , [4]–[9]. , - (LVI) [8]. , , . (TB) , , . , (EMT)[10], [11]. , , EMT , , . [10], [12]– [16]. , 30% Georgian Scientists/ 4(02), 2022 120 . , , . (FFPE) 55 Luminal A 33 22 – Luminal B. , . 2017-2021 . 1-3): ; ; St. Gallen 2013 criteria [18]; ; ki67; E- , , Her2 ( Antibody Clones: ER–clone:6F11; PR-clone:16; HER2-clone: CB11; Ki67-clone:MM1; Vimentin – clone:V9; E-cadherin – clone:36B5; Beta-Catenin Clone:17C2; - NovolinkTM Max Polymer Detection System). . H&E QuPath. (TB - ) H&E ( ; ). H&E , Qupath- Quantitative Pathology & Bioimage Analysis 0.3.2. [19] 1 2). Georgian Scientists/ 4(02), 2022 121 Georgian Scientists/ 4(02), 2022 122 ,, ’’ 2016 International Tumor Budding Consensus Conference (ITBCC) 2016[20]. (TB) : : 5 [20]. H&E ( ; ). : Spearman’s rank test - Mann-Whitney Kruskal-Wallis . 95% . P <0.05 . SPSS statistical software V20.0- . Georgian Scientists/ 4(02), 2022 123 Luminal A 33 Grade 1 3 (9,1%); Grade 2 – 11 (33.3%); Grade 3 – 19 (57.6%); Luminal A 33 0-10%- 7 (21.2%); 11-50% - 12 (36.4%), >51% - 14 (42.4%); 0-10%- 10 (30.3%); 11-50%- 12 (36.4); >51%> 14 (42.4%); 0-10% 6 (18.2%); 11-50%- >51%- . 0-10%- 7 (21.2%); 11-50% >51%- ; KI-67 1-3% 33 3 (9.1%), 4-9% 8 (24.2%); 9-14% 22 (66.7%), >14 % . Luminal B 22 Grade 1 3 (13.6%); Grade 2 -12 (54.5%); Grade 3 – 7 (31.8%); Luminal B 22 0-10%- 5 (22.7%); 11-50% - 9 (40.9%), >51% - 8 (36.4%); 0-10%- 4 (18.2%); 11-50%- 11 (50.0%); >51%> 7 (31.8%); 0-10% 9 (40.9%); 11-50%- >51%- . 0-10%- 11 (50.0%); 11-50% >51%- ; Ki67- 1-3% , 4-9% 1 (4.5%) 9- 14% 3 (13.6%) 18 >14% 1). Georgian Scientists/ 4(02), 2022 124 1: (n=55) Table 1 Luminal A Luminal B N % N % Luminal 33.0 60.0 22.0 40.0 Nottingham Grade Grade 1 3 9.1 3 13.6 Grade 2 11 33.3 12 54.5 Grade 3 19 57.6 7 31.8 ER - 0-10% 7 21.2 5 22.7 11-50% 12 36.4 9 40.9 >51% 14 42.4 8 36.4 PR - 0-10% 10 30.3 4 18.2 11-50% 12 36.4 11 50.0 >51% 11 33.3 7 31.8 0-10% 6 18.2 9 40.9 11-50% 0 0.0 0 0.0 >51% 0 0.0 0 0.0 0-10% 7 21.2 11 50.0 11-50% 0 0.0 0 0.0 >51% 0 0.0 0 0.0 Ki67 1-3% 3 9.1 0 0.0 4-9% 8 24.2 1 4.5 9-14% 22 66.7 3 13.6 14 >% 0 0.0 18 81.8 Luminal A 33 (TB: 0-5) 6/33 (18.2%); 6 11-50 % 2 (33.3%); 51% 4 (66.7%); 11-50% 3 (50.0%); 51%> 3 (50.0%). 0-1% 2 (33.3%); 1-10%- 3 (50.0%); 11-50%- - 1 (16.7%); 0-1% 3 (50.0%); 1-10% 2 (33.3%); 11-50% - 1 (16.7); E- 11-50% 2 (33.3); 51%> 4 (66.7%); Ki67- 4-9%- 1 (16.7%); 9-14%- 5 (83.3%); Georgian Scientists/ 4(02), 2022 125 (TB: 5-9) 18/33 (54.5%); 18 11-50% 7 (38.9%); 51%> 11 (61.1%); 11-50% 10 (55.6%) >51% 8 (44.4%); 0-1% 4 (22.2%); 1-10% 11 (61.1%) 11-50% 3 (16.7%) >51% ; 0-1% 5 (27.8%) 1-10% 9 (50.0%); 11-50% 4 (22.2%), >51%- . E- 18 11-50% 8 (44.4%) >51%- 10 (55.6%), 1-10% 0-1% E- . Ki67- 18 1-3% 3 (16.7%), 4-9% - 6 (33.3%) 9-14% -9 (50.0%) <14%- . (TB: >9) 9/33 (27.3%); 9 1-10% 6 (66.7%) 11-50% 3 (33.3%) >51% . 1-10% 4 (44.4%) 11-50% 5 (55.6) >51% . 0-1% 2 (22.2%), 1-10% 3 (33.3%) 11-50%% - 4 (44.4%) >51% . 0-1% 1 (11.1%) 1-10% 2 (22.2%), 11-50% - 6 (66.7%) >51% . E- 0-1% 1-10% . 11-50% 8 (44.4%) >51% - 10 (55.6%). Ki 67- 1-3%- 4 (44.4%) 4-9% - 2 (22.2%), 9-14% 2 (22.2%) Ki67- >14%- . Luminal B 22 (TB :0-5) 3/22 (13.6%); 1- Georgian Scientists/ 4(02), 2022 126 10% 2 (66.7%), 11-50% 1 (33.3%) >51% . 1-10% 1 (33.3%), 11-50% 2 (66.7%), >51%- . 0-1% 1 (33.3%), 11-50% 2 (66.7%) 1-10% >51% . 0-1% 1 (33.3%), 1-10% 1 (33.3%), 11- 50% - 1 (33.3) >51%- . E- 11-50% 3 (100%) 0- 1%, 1-10% >51% E- . Ki67- 9-14% 1 (33.3%) >14%- 2 (66.7%) 4-9% 1-3%- . (TB: 5-9) 5/22 (22.7%); 1-10% 3 (60%), 11-50% 2 (40.0%) >51- . 1-10% 2 (40%) 11- 50% 3 (60.0%) >51% . 0-1% 1 (20.0%) 11-50% 4 (80%) 1-10 >51% . 0-1% 1 (20%) 1-10% 1 (20%) 11-50% 3 (60%) >51% . E- 1-10% 1 (20%) 11-50% 4 (80%) 0-1% >51% E- . Ki67- 9-14% 2 (40%), >14% 3 (60%) 1-3% 4-9% (TB: >9) 14/22 (63.6%); 1-10% 10 (71.4%) , 11-50% 4 (28.6%), >51% . 1-10% 9 (64.3%), 11-50% 5 (35.7%), >51% . Georgian Scientists/ 4(02), 2022 127 0-1% 2 (14.3%) 11-50% 5 (35.7%) >51%- - 7 (50.0%). 1-10% . 0-1% 2 (14.3%) 11-50% 3 (21.4%) >51% 9 (64.3%). 1-10% . E- 0-1% 2 (14.3%) 1-10% 5 (35.7%) 11-50%- 7 (50.0%) >51% . Ki-67 4-9% 2 (14.3%) 9-14%- 4 (28.6%) >14% 8 (57.1%) 2). 2: ; Luminal A - 33 Luminal B - 22 TB TB 0-5 Low 5-9 (Medium) >9 (High) 0-5 Low 5-9 (Medium) >9 (High) N/Frequency 6 18.2 18 54.5 9 27.3 3 13.6 5 22.7 14 63.6 ER 1-10% 0 0 0 0 6 66.7 2 66.7 3 60 10 71.4 11-50% 2 33.3 7 38.9 3 33.3 1 33.3 2 40 4 28.6 51%> 4 66.7 11 61.1 0 0 0 0 0 0 0 0 PR 1-10% 0 0 0 0 4 44.4 1 33.3 2 40 9 64.3 11-50% 3 50 10 55.6 5 55.6 2 66.7 3 60 5 35.7 51%> 3 50 8 44.4 0 0 0 0 0 0 0 0 0-1% 2 33.3 4 22.2 2 22.2 1 33.3 1 20 2 14.3 1-10% 3 50 11 61.1 3 33.3 0 0 0 0 0 0 11-50% 1 16.7 3 16.7 4 44.4 2 66.7 4 80 5 35.7 51%> 0 0 0 0 0 0 0 0 0 0 7 50 - 0-1% 3 50 5 27.8 1 11.1 1 33.3 1 20 2 14.3 1-10% 2 33.3 9 50 2 22.2 1 33.3 1 20 0 0 11-50% 1 16.7 4 22.2 6 66.7 1 33.3 3 60 3 21.4 51%> 0 0 0 0 0 0 0 0 0 0 9 64.3 0-1% 0 0 0 0 0 0 0 0 0 0 2 14.3 E- 1-10% 0 0 0 0 3 33.3 0 0 1 20 5 35.7 11-50% 2 33.3 8 44.4 5 55.6 3 100 4 80 7 50 51%> 4 66.7 10 55.6 0 0 0 0 0 0 0 0 Ki67 1-3% 0 0 3 16.7 4 44.4 0 0 0 0 0 0 4-9% 1 16.7 6 33.3 2 22.2 0 0 0 0 2 14.3 9-14% 5 83.3 9 50 2 22.2 1 33.3 2 40 4 28.6 14 >% 0 0 0 0 0 0 2 66.7 3 60 8 57.1 Georgian Scientists/ 4(02), 2022 128 Luminal A 33 (TB: 0- 5) 6 1-10% 1 (16.7%) 11-50% 2 (33.3%) >51% 3 (50.05%) 0-1% . (TB: 5-9) 18 1-10% 3 (16.7%), 11-50% - 6 (33.3%) >51% 9 (50.0) 0-1% . (TB: >9) 9 0-1% 2 (22.2%), 1-10% 6 (66.7%) 11-50% 1 (11.1%) >51% . Luminal B 22 (TB: 0- 5) 3 1-10% (100.%) 0-1%; 11-50% >51% . (TB: 5-9) 5 1-10% 4 (80%) 11-50% 1 (20%). 0-1% >51% . (TB: >9) 14 0-1% 3 (21.4%) 1-10% 9 (64.3%) 11-50% 2 (14.3%). >51% . Luminal A (TB: 0-5) 6 (6/33) 1-10% 1 (16.7%) 11-50% 3 (50.0%), >51% 2 (33,3), 0-1% . (TB: 5-9) 18 (18/33) Georgian Scientists/ 4(02), 2022 129 0-1% 1 (5.56%), 1-10% 3 (16.7%), 11-50% 8 44,4%, >51%- 6 (33,3%). (TB: >9) 9 (9/33) 0-1% 1 (11.1%), 1-10% 5 (55.6%), 11-50% 3 (33,3%), >51% . Luminal B (TB: 0-5) 3 (3/22) 1-10% 2 (66,7%) 11-50% 1 (33,3%), 0-1% >51% . (TB: 5-9) 5 (5/22) 1-10% 3 (60.0%) 11-50% 2 (40.0%), 0-1% >51%- . (TB: >9) 14 0-1% 3 (21.4%) 1-10% 8 (57.1%) , 11-50% 3 (21.4%) %), 0-1% >51%- . Luminal A (TB: 0-5) 6 (6/33) 0-1% 4 (66.7%), 1-10% 2 (33.3%), 11-50% >51% . (TB: 5-9) 18 (18/33) 0-1% 8 (44.4%) 1-10% 7 (38.9%), 11-50% 3 (16.7%) >51% . (TB: >9) 9 (9/33) 0-1% 4 (44.4%), 1-10% 4 (44.4%), 11-50% 1 (11.1%) >51% . Georgian Scientists/ 4(02), 2022 130 Luminal B (TB: 0-5) 3 (3/22) 0-1% 2 (66.7%), 1-10% 1 (33.3%) 11-50% >51% . (TB: 5-9) 5 (5/22) 0-1% 3 (60%) 1-10% 2 (40%) 11-50% >51% . (TB: >9) 14 0-1% 4 (28,6%) 1-10% 5 (35.7%), 11-50% 5 (35,7%) >51%- . Luminal A (TB: 0-5) 6 (6/33) - 0- 1% 2 (33.3%) 1-10% 3 (50%), 11- 50% 1 (16.7%), <51% . (TB: 5-9) 18 (18/33) 0-1% 7 (38.9%), 1-10% 7 (38.9%), 11-50% 4 (22.2%), >51% . (TB: >9) 9 (9/33) 0-1% 4 (44.4%), 1-10% 3 (33.3%), 11-50% 2 (22.2%) >51% . Luminal B (TB: 0-5) 3 (3/22) 0-1% 3 (100%). 1-10% , 11-50% >51% . (TB: 5-9) 5 (5/22) - 0-1% 3 (60%), 1-10% 1 (20%), 11-50% 1 (20%), >51 . (TB: >9) 14 - Georgian Scientists/ 4(02), 2022 131 0-1% 6 (42.9%), 1-10% 4 (28.6%), 11-50% 2 (14.3%) >51% 2 (14.3%). Luminal A (TB: 0-5) 6 (6/33) E- 11- 50% 1 (16.7%) >51% 5 (83.3%) 0-1% 1-10% . (TB: 5-9) 18 (18/33) E- 11-50% 9 (50%), >51% 9 (50%) 0-1% 1-10% . (TB: >9) 9 (9/33) E- 1-10% 1 (11.1%) 11-50% 6 (66.7%) >51% 2 (22.2%) 0-1% . Luminal B (TB: 0-5) 3 (3/22) E- 11-50% 2 (66.7) >51% 1 (33,3%) 0- 1% 1-10% . (TB: 5-9) 5 (5/22) E- 11-50% 4 (80%), >51% 1 (20%) 0-1% 1-10% . (TB: >9) 14 (14/22) E- 1-10% 3 (21.4%) 11-50% 9 (64.3%) >51% 2 (14.3%) 0-1% 3). Georgian Scientists/ 4(02), 2022 132 3: Luminal A - 33 Luminal B - 22 TB score 0-5 Low 5-9 (Medium) >9 (High) 0-5 Low 5-9 (Medium) >9 (High) N/Frequency 6 18.2 18 54.5 9 27.3 3 13.6 5 22.7 14 63.6 0-1% 0 0 0 0 2 22.2 0 0 0 0 3 21.4 ER 1-10% 1 16.7 3 16.7 6 66.7 3 100 4 80 9 64.3 11- 50% 2 33.3 6 33.3 1 11.1 0 0 1 20 2 14.3 51%> 3 50 9 50 0 0 0 0 0 0 0 0 0-1% 0 0 1 5.56 1 11.1 0 0 0 0 3 21.4 PR 1-10% 1 16.7 3 16.7 5 55.6 2 66.7 3 60 8 57.1 11- 50% 3 50 8 44.4 3 33.3 1 33.3 2 40 3 21.4 51%> 2 33.3 6 33.3 0 0 0 0 0 0 0 0 Vimentin 0-1% 4 66.7 8 44.4 4 44.4 2 66.7 3 60 4 28.6 1-10% 2 33.3 7 38.9 4 44.4 1 33.3 2 40 5 35.7 11- 50% 0 0 3 16.7 1 11.1 0 0 0 0 5 35.7 51%> 0 0 0 0 0 0 0 0 0 0 0 0 - 0-1% 2 33.3 7 38.9 4 44.4 3 100 3 60 6 42.9 1-10% 3 50 7 38.9 3 33.3 0 0 1 20 4 28.6 11- 50% 1 16.7 4 22.2 2 22.2 0 0 1 20 2 14.3 51%> 0 0 0 0 0 0 0 0 0 0 2 14.3 0-1% 0 0 0 0 0 0 0 0 0 0 0 0 E- 1-10% 0 0 0 0 1 11.1 0 0 0 0 3 21.4 11- 50% 1 16.7 9 50 6 66.7 2 66.7 4 80 9 64.3 51%> 5 83.3 9 50 2 22.2 1 33.3 1 20 2 14.3 , Luminal A , (11-50 <51%), Luminal B Georgian Scientists/ 4(02), 2022 133 ( N1). 0- 1% 1- 10% 11- 50% 51%> A - 1 21 36 42 B - 1 23 41 36 A - TB: 1 1 33 67 A - TB: 1 1 39 61 A - TB: 1 67 33 1 B - TB: 1 67 33 1 B - TB: 1 60 40 1 B - TB: 1 71 29 1 A - TB: / 1 17 33 50 A - TB: / 1 17 33 50 A - TB: / 22 67 11 1 B - TB: / 1 100 1 1 B - TB: / 1 80 20 1 B - TB: / 21 64 14 1 Luminal A Luminal B , . , (TB) 96.5%- , 0 0 0 0 0 0 0 0 0 0 22 0 0 2121 23 0 0 67 67 60 71 17 17 67 100 80 64 36 41 33 39 33 33 40 29 33 33 11 0 20 14 42 36 67 61 0 0 0 0 50 50 0 0 0 0 0 10 20 30 40 50 60 70 80 90 100 A - B - A -T B: A -T B: A -T B: B -T B: B -T B: B -T B: A -T B: / A -T B: / A -T B: / B -T B: / B -T B: / B -T B: / 1: ER- Luminal A B , (TB) 0-1% 1-10% 11-50% 51%> Georgian Scientists/ 4(02), 2022 134 Luminal A Luminal B [6 ]. Luminal A . Luminal B ( N2). 0- 1% 1- 10% 11- 50% 51%> A - 1 18 1 1 B - 1 41 1 1 A - TB: 33 50 17 1 A - TB: 22 61 17 1 A - TB: 22 33 44 1 B - TB: 33 1 67 1 B - TB: 1 80 1 20 B - TB: 14 1 36 50 A - TB: / 67 33 1 1 A - TB: / 44 39 17 1 A - TB: / 44 44 11 1 B - TB: / 67 33 1 1 B - TB: / 60 40 1 1 B - TB: / 29 36 36 1 0 0 33 22 22 33 0 14 67 44 44 67 60 29 18 41 50 61 33 0 80 0 33 39 44 33 40 36 0 0 17 17 44 67 0 36 1 17 11 1 1 36 0 0 0 0 0 0 20 50 1 1 1 1 1 1 0 10 20 30 40 50 60 70 80 90 100 A - B - A -T B: A -T B: A -T B: B -T B: B -T B: B -T B: A -T B: / A -T B: / A -T B: / B -T B: / B -T B: / B -T B: / 2: Luminal A B , (TB) 0-1% 1-10% 11-50% 51%> Georgian Scientists/ 4(02), 2022 135 Luminal A 2/3- Luminal B- . . [21] Luminal A . Luminal B . 1/3- . Luminal B 100% 30%- ( N3). 0- 1% 1- 10% 11- 50% 51%> A - 1 21 1 1 B - 1 50 1 1 A - TB: 50 33 17 1 A - TB: 28 50 22 1 A - TB: 11 22 67 1 B - TB: 33 33 33 1 B - TB: 20 20 60 1 B - TB: 14 1 21 64 A - TB: / 33 50 17 1 A - TB: / 39 39 22 1 A - TB: / 44 33 22 1 B - TB: / 100 1 1 1 B - TB: / 60 20 20 1 B - TB: / 43 29 14 14 Georgian Scientists/ 4(02), 2022 136 E- . 15% N4). 0- 1% 1- 10% 11- 50% 51%> A - TB: 1 1 33 67 A - TB: 1 1 44 56 A - TB: 1 33 56 1 B - TB: 1 1 100 1 B - TB: 1 20 80 1 B - TB: 14 36 50 1 A - TB: / 1 1 17 83 A - TB: / 1 1 50 50 A - TB: / 1 11 67 22 B - TB: / 1 1 67 33 B - TB: / 1 1 80 20 B - TB: / 1 21 64 14 0 0 50 28 11 33 20 14 33 39 44 100 60 43 21 50 33 50 22 33 20 0 50 39 33 0 20 29 0 0 17 22 67 33 60 21 17 22 22 0 20 14 0 0 0 0 0 0 0 64 0 0 0 0 0 14 0 10 20 30 40 50 60 70 80 90 100 A - B - A -T B: A -T B: A -T B: B -T B: B -T B: B -T B: A -T B: / A -T B: / A -T B: / B -T B: / B -T B: / B -T B: / 3: Luminal A B , (TB) 0-1% 1-10% 11-50% 51%> Georgian Scientists/ 4(02), 2022 137 Luminal B . , . , , . 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Lugli et al., “Recommendations for reporting tumor budding in colorectal cancer based on the International Tumor Budding Consensus Conference (ITBCC) 2016,” Modern Pathology 2017 30:9, vol. 30, no. 9, pp. 1299–1311, May 2017, doi: 10.1038/modpathol.2017.46. [21] K. A. Bhuvaneshwari, “Expression of vimentin in breast carcinoma and its correlation with histopathological parameters,” Indian Journal of Pathology and Oncology Journal homepage: www.ijpo.co.in Original Research Article, vol. 8, no. 2, pp. 207–212, 2021, doi: 10.18231/j.ijpo.2021.041. Georgian Scientists/ 4(02), 2022 140 Comparative analysis of phenotypic features in the primary tumor, tumour buds and metastatic lymph nodes within Luminal A and Luminal B molecular subtypes of invasive ductal carcinoma of the breast Shota Kepuladze1, Irakli Kokhreidze2, George Burkadze3 1Tbilisi State Medical University PhD student, MD 2Associated Professor MD., PhD. VICE RECTOR of Tbilisi State Medical University Head of Oncology Department of TSMU First University Clinic 3Professor at Tbilisi State Medical University, Head of the Department of Molecular pathology Abstract The study included 55 cases of formalin-fixed and paraffin-embedded (FFPE) tissues of breast invasive ductal carcinoma. The following algorithm has been made for further discussion by using immunohistochemical examination: antibodies against hormonal receptors; HER2; E-cadherin, Vimentin, Beta-catenin; Ki-67; Tumour buds were evaluated by using H&E stained slides and computer software Qupath (version 0.3.2). The results of the study show that estrogen expression is different in primary tumour mass and in tumour buds and its expression is diminished in the Luminal B molecular subtype respectively. Vimentin and Beta-catenin expression is showing similar changes, the quantity is much higher in tumour microclusters compared to the primary tumour and metastatic lymph nodes. It can demonstrate features of epithelial-mesenchymal transformation. Modifications in dynamics of proliferative activity are showing the lowest proliferative activity in tumour microclusters which can be discussed as the indirect manifestation of epithelial-mesenchymal transformation. Key Words: Breast Cancer; Epithelia-mesenchymal transition; Tumor buds;