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Tumor Discovery Colorectal cancer: miRNA, mRNA, protein insights

Table 2. (Continued) Table 2. (Continued)
Variable miRNA expression P P‑value of Variable miRNA expression P P‑value of
Normal (%) High (%) Bonferroni Normal (%) High (%) Bonferroni
correction test correction test
Nodal state C 19 (33.3) 12 (50.0)
pN 0 22 (61.1) 28 (62.2) 0.6 0.9 EMVI
pN I 12 (33.3) 12 (26.7) 0 31 (54.4) 10 (41.7) 0.3 0.7
pN II 2 (5.6) 5 (11.1) 1 24 (42.1) 14 (58.3)
Dukes’ stage 2 2 (3.5) 0 (0.0)
A 4 (11.1) 8 (17.8) 0.6 0.9 miR-224
B 18 (50.0) 20 (44.4) Tumor grade
C 14 (38.9) 17 (37.8) Well 1 (2.3) 1 (2.7) 0.4 0.8
EMVI Good 39 (88.6) 35 (94.6)
0 19 (52.8) 22 (48.9) 0.4 0.8 Poor 4 (9.1) 1 (2.7)
1 17 (47.2) 21 (46.7) Nodal state
2 0 (0.0) 2 (4.4) pN 0 29 (65.9) 21 (56.8) 0.6 0.9
miR-31 pN I 11 (25.0) 13 (35.1)
Tumor grade pN II 4 (9.1) 3 (8.1)
Well 1 (2.9) 1 (2.1) 0.1 0.3 Dukes’ stage
Good 29 (85.3) 45 (95.7) A 6 (13.6) 6 (16.2) 0.5 0.9
Poor 4 (11.8) 1 (2.1) B 23 (52.3) 15 (40.5)
Nodal state C 15 (34.1) 16 (43.2)
pN 0 22 (64.7) 28 (59.6) 0.1 0.3 EMVI
pN I 7 (20.6) 17 (36.2) 0 23 (52.3) 18 (48.6) 0.9 0.9
pN II 5 (14.7 2 (4.3) 1 20 (45.5) 18 (48.6)
Dukes’ stage 2 1 (2.3) 1 (2.7)
A 2 (5.9) 10 (21.3) 0.1 0.3 Abbreviation: EMVI: Extramural vascular invasion.
B 19 (55.9) 19 (40.4)
C 13 (38.2) 18 (38.3) expected bands for each antibody in specific cell lysates:
EMVI SMAD4 in SW480, RASA1 in HT29 and Lovo, KLF4 in
0 14 (41.2) 27 (57.4) 0.3 0.7 HT29, and TGFBRII in HT29 and SW480 (Figure S1
1 19 (55.9) 19 (40.4) in Supplementary File). These results validated the
specificity of the antibodies for staining target proteins
2 1 (2.9) 1 (2.1) within CRC TMAs via immunohistochemistry. Other
miR-92a antibodies used in our study, sourced from our group and
Tumor grade the histopathological department at Nottingham QMC,
Well 1 (1.8) 1 (4.2) 0.03 0.2 underwent similar validation procedures. In addition,
Good 55 (96.5) 19 (79.2) we optimized the concentration of each antibody for
Poor 1 (1.8) 4 (16.7) immunohistochemical staining. For example, anti-
Nodal state SMAD4 at 1:100 exhibited optimal staining without
pN 0 38 (66.7) 12 (50.0) 0.2 0.6 background, while concentrations of 1:50 and 1:200
were either too high or too low for detection. Similarly,
pN I 14 (24.6) 10 (41.7) optimal concentrations were determined for TGFBRII
pN II 5 (8.8) 2 (8.3) (1:400), RASA1 (1:40), and KLF4 (1:100). Staining
Dukes’ stage for BCL2 and PTEN was performed separately by the
A 9 (15.8) 3 (12.5) 0.3 0.7 histopathology department at QMC. Further details
B 29 (50.9) 9 (37.5) on the staining optimization process can be found in
(Cont’d...) Table 1 and Figure S2 (in Supplementary file).

Volume 4 Issue 1 (2025) 73 doi: 10.36922/td.4631