Background
Methods
Patients and tissue samples
Radiotherapy
RNA isolation and cDNA synthesis
Global microRNA profiling
Validation of the differentially expressed microRNAs
Statistical analysis
Results
Characteristics of the patients
Number of cases (%) | Radiosensitive | Radioresistant | X2 (p value) | |
---|---|---|---|---|
n (%) | n (%) | |||
Gender | ||||
Male | 30 (88.2) | 12 (40.0) | 18 (60.0) | 1.0 |
Female | 4 (11.8) | 2 (50.0) | 2 (50.0) | |
Age | ||||
≤60 years | 16 (47.1) | 5 (31.2) | 11 (68.8) | 0.51 |
>60 years | 18 (52.9) | 9 (50.0) | 9 (50.0) | |
Tobacco consumption | ||||
Yes | 31 (91.2) | 14 (45.2) | 17 (54.8) | 0.25 |
No | 3 (8.8) | 0 (0) | 3 (100.0) | |
Tumor site | ||||
Supraglottic | 7 (20.6) | 2 (28.6) | 5 (71.4) | 1.0 |
Glottic | 27 (79.4) | 12 (44.4) | 15 (55.6) | |
Tumor stage | ||||
I | 16 (47.1) | 8 (50) | 8 (50) | 0.49 |
II | 18 (52.9) | 6 (33.3) | 12 (66.7) | |
Radiotherapy dose | ||||
<70 Gy | 10 (29.4) | 5 (50.0) | 5 (50.0) | 0.7 |
≥70 Gy | 24 (70.6) | 9 (37.5) | 15 (62.5) | |
Treatment time | ||||
≤55 days | 20 (58.8) | 9 (45.0) | 11 (55.0) | 0.72 |
>55 days | 14 (41.2) | 5 (37.5) | 9 (64.3) |
MicroRNA expression in LSCC samples
Up-regulated | Down-regulated | ||||
---|---|---|---|---|---|
microRNA | Fold-change | FDR-adjusted p-value | microRNA | Fold-change | FDR-adjusted p value |
hsa-miR-642 | 2.02 | 0.3869 | hsa-miR-106b | 2.01 | 0.2162 |
hsa-miR-744 | 2.02 | 0.4254 | hsa-miR-302b | 2.05 | 0.5657 |
hsa-miR-221 | 2.03 | 0.3475 | hsa-miR-136* | 2.06 | 0.5306 |
hsa-miR-193b* | 2.07 | 0.3359 | hsa-miR-127-5p | 2.09 | 0.5626 |
hsa-miR-193b | 2.08 | 0.4149 | hsa-miR-152 | 2.09 | 0.3678 |
hsa-miR-18b | 2.11 | 0.3012 | hsa-miR-34c-5p | 2.09 | 0.4936 |
hsa-miR-222*- | 2.15 | 0.4874 | hsa-miR-199a-5p | 2.11 | 0.6033 |
hsa-miR-944 | 2.16 | 0.4246 | hsa-miR-190b | 2.12 | 0.6416 |
hsa-miR-766- | 2.21 | 0.3723 | hsa-miR-140-3p | 2.14 | 0.4081 |
hsa-miR-106b* | 2.22 | 0.3155 | hsa-let-7i* | 2.16 | 0.5682 |
hsa-miR-942 | 2.35 | 0.3275 | hsa-miR-195 | 2.17 | 0.508 |
hsa-miR-135a | 2.41 | 0.1506 | hsa-miR-302c | 2.17 | 0.5853 |
hsa-miR-652 | 2.41 | 0.2572 | hsa-miR-488 | 2.17 | 0.5469 |
hsa-miR-124 | 2.42 | 0.3872 | hsa-miR-411* | 2.17 | 0.5271 |
hsa-miR-105 | 2.51 | 0.2277 | hsa-miR-21 | 2.18 | 0.3333 |
hsa-miR-923 | 2.62 | 0.5362 | hsa-miR-590-5p | 2.18 | 0.37 |
hsa-miR-138 | 2.71 | 0.2162 | hsa-miR-29c | 2.23 | 0.4804 |
hsa-miR-181a-2* | 2.78 | 0.2034 | hsa-miR-454 | 2.24 | 0.3152 |
hsa-miR-23b | 2.84 | 0.2235 | hsa-miR-10b* | 2.25 | 0.5091 |
hsa-miR-224 | 2.85 | 0.2626 | hsa-miR-378* | 2.29 | 0.4961 |
hsa-miR-141 | 2.89 | 0.3926 | hsa-miR-380* | 2.31 | 0.4251 |
hsa-miR-205 | 3.13 | 0.2706 | hsa-miR-661 | 2.31 | 0.5051 |
hsa-miR-129-3p | 3.53 | 0.1788 | hsa-miR-10a | 2.34 | 0.1452 |
hsa-miR-188-5p | 3.58 | 0.5457 | hsa-miR-148b* | 2.34 | 0.4926 |
hsa-miR-107 | 3.81 | 0.1843 | hsa-miR-592 | 2.35 | 0.4694 |
hsa-miR-183 | 3.91 | 0.1201 | hsa-miR-449b | 2.36 | 0.526 |
hsa-miR-296-5p
|
4.26
|
0.084
| hsa-miR-487a | 2.36 | 0.511 |
hsa-miR-23a
|
4.58
|
0.1771
| hsa-miR-199a-3p | 2.37 | 0.4496 |
hsa-miR-22
|
4.78
|
0.3268
| hsa-miR-181a | 2.41 | 0.3499 |
hsa-miR-801
|
5.87
|
0.127
| hsa-miR-30a* | 2.48 | 0.3854 |
hsa-miR-200a
|
6.22
|
0.3678
| hsa-miR-144* | 2.51 | 0.5976 |
hsa-miR-203
|
6.22
|
0.1755
| hsa-miR-509-3p | 2.56 | 0.4307 |
hsa-miR-429
|
6.68
|
0.1588
| hsa-miR-146b-5p | 2.57 | 0.224 |
hsa-miR-183*
|
7.84
|
0.0203
| hsa-miR-154* | 2.57 | 0.3571 |
hsa-miR-200b*
|
8.02
|
0.1744
| hsa-miR-638 | 2.61 | 0.4688 |
hsa-miR-601
|
8.31
|
0.2335
| hsa-miR-328 | 2.61 | 0.3059 |
hsa-miR-200b
|
9.41
|
0.1332
| hsa-miR-656 | 2.62 | 0.4201 |
hsa-miR-200c
|
9.77
|
0.1334
| hsa-miR-146b-3p | 2.68 | 0.328 |
hsa-miR-200a*
|
9.86
|
0.0974
| hsa-miR-202 | 2.77 | 0.3485 |
hsa-miR-452
|
17.94
|
0.0238
| hsa-miR-548b-5p | 2.77 | 0.4288 |
hsa-miR-650
|
18.46
|
0.1454
| hsa-miR-340 | 2.78 | 0.2592 |
hsa-miR-378
|
28.01
|
0.3958
| hsa-miR-15a* | 2.81 | 0.2874 |
hsa-miR-513-3p
|
229.63
|
0.355
| hsa-miR-29c* | 2.87 | 0.4847 |
hsa-miR-29a* | 2.89 | 0.3125 | |||
hsa-miR-132 | 2.91 | 0.2681 | |||
hsa-miR-345 | 2.98 | 0.0946 | |||
hsa-miR-214 | 3.02 | 0.4376 | |||
hsa-miR-376a | 3.05 | 0.4487 | |||
hsa-miR-20a* | 3.11 | 0.183 | |||
hsa-miR-539 | 3.12 | 0.4698 | |||
hsa-miR-487b | 3.19 | 0.4356 | |||
hsa-miR-433 | 3.21 | 0.3942 | |||
hsa-miR-639 | 3.21 | 0.3717 | |||
hsa-miR-885-5p | 3.27 | 0.3696 | |||
hsa-miR-520c-3p | 3.28 | 0.3564 | |||
hsa-miR-127-3p | 3.29 | 0.4056 | |||
hsa-miR-99a* | 3.32 | 0.2645 | |||
hsa-miR-361-5p | 3.39 | 0.1345 | |||
hsa-miR-134 | 3.41 | 0.369 | |||
hsa-miR-130b* | 3.51 | 0.3143 | |||
hsa-miR-493 | 3.53 | 0.3315 | |||
hsa-miR-212 | 3.58 | 0.241 | |||
hsa-miR-19b-1* | 3.68 | 0.3595 | |||
hsa-miR-30d* | 3.74 | 0.091 | |||
hsa-miR-760 | 3.75 | 0.4218 | |||
hsa-miR-125b-1* | 3.77 | 0.3397 | |||
hsa-miR-140-5p | 3.81 | 0.1951 | |||
hsa-miR-34b* | 3.97 | 0.4022 | |||
hsa-miR-99b*
|
4.17
|
0.214
| |||
hsa-miR-889
|
4.17
|
0.294
| |||
hsa-miR-516a
|
4.17
|
0.244
| |||
hsa-miR-770
|
4.17
|
0.274
| |||
hsa-miR-671
|
4.55
|
0.174
| |||
hsa-miR-100
|
4.76
|
0.204
| |||
hsa-miR-543
|
4.76
|
0.264
| |||
hsa-miR-410
|
5.00
|
0.285
| |||
hsa-miR-495
|
5.00
|
0.385
| |||
hsa-miR-485
|
5.26
|
0.325
| |||
hsa-miR-411
|
5.56
|
0.235
| |||
hsa-miR-16-1*
|
5.56
|
0.045
| |||
hsa-miR-382
|
5.88
|
0.345
| |||
hsa-miR-100*
|
5.88
|
0.225
| |||
hsa-miR-7-1*
|
5.88
|
0.505
| |||
hsa-miR-337
|
6.25
|
0.756
| |||
hsa-miR-451
|
6.25
|
0.236
| |||
hsa-miR-99a
|
7.14
|
0.167
| |||
hsa-miR-218
|
7.69
|
0.847
| |||
hsa-miR-370
|
7.69
|
0.327
| |||
hsa-miR-379
|
8.33
|
0.788
| |||
hsa-miR-409
|
9.09
|
0.369
| |||
hsa-miR-376c
|
10.00
|
0.1110
| |||
hsa-miR-432
|
11.11
|
0.2111
| |||
hsa-miR-214*
|
12.50
|
0.1412
|
miR-296-5p expression is associated with patient prognosis
Variable | Categories | Number of cases | miR-296-5p | miR-452 | miR-183* | miR-200c | ||||
---|---|---|---|---|---|---|---|---|---|---|
Low | High | Low | High | Low | High | Low | High | |||
n (%) | n (%) | n (%) | n (%) | n (%) | n (%) | n (%) | n (%) | |||
Age (years) | ≤60 years | 16 | 6 (35.3) | 10 (58.8) | 8 (47.1) | 8 (47.1) | 8 (47.1) | 8 (47.1) | 8 (50.0) | 8 (47.1) |
>60 years | 18 | 11 (64.7) | 7 (41.2) | 9 (52.9) | 9 (52.9) | 9 (52.9) | 9 (52.9) | 8 (50.0) | 9 (52.9) | |
p (2-sided) | NS | NS | NS | NS | ||||||
Gender | Female | 4 | 2 (11.8) | 2 (11.8) | 2 (11.8) | 2 (11.8) | 2 (11.8) | 2 (11.8) | 2 (12.5) | 2 (12.5) |
Male | 30 | 15 (88.2) | 15 (88.2) | 15 (88.2) | 15 (88.2) | 15 (88.2) | 15 (88.2) | 14 (87.5) | 15 (88.2) | |
p (2-sided) | NS | NS | NS | NS | ||||||
Smoking | No | 3 | 0 (0) | 3 (17.6) | 1 (5.9) | 2 (11.8) | 0 (0) | 3 (17.6) | 2 (12.5) | 1 (9.1) |
Yes | 31 | 17 (100) | 14 (82.4) | 16 (94.1) | 15 (88.2) | 17 (100) | 14 (82.4) | 14 (87.5) | 16 (94.1) | |
p (2-sided) | NS | NS | NS | NS | ||||||
Tumor site* | Glottic | 27 | 15 (88.2) | 12 (70.6) | 14 (82.4) | 13 (76.5) | 14 (82.4) | 13 (76.5) | 14 (87.5) | 12 (70.6) |
Supraglottic | 7 | 2 (11.8) | 5 (29.4) | 3 1 (7.6) | 4 (23.5) | 3 (17.6) | 4 (23.5) | 2 (12.5) | 5 (29.4) | |
p (2-sided) | NS | NS | NS | NS | ||||||
Tumor stage | I | 16 | 7 (41.2) | 9 (52.9) | 9 (52.9) | 7 (41.2) | 7 (41.2) | 9 (52.9) | 7 (43.8) | 9 (52.9) |
II | 18 | 10 (58.8) | 8 (47.1) | 8 (47.1) | 10 (58.8) | 10 (58.8) | 8 (47.1) | 9 (56.2) | 8 (47.1) | |
p (2-sided) | NS | NS | NS | NS | ||||||
Radiosensibility | Sensivite | 14 | 11 (64.7) | 3 (17.6) | 8 (47.1) | 6 (35.3) | 9 (52.9) | 5 (29.4) | 7 (43.8) | 7 (41.2) |
Resistant | 20 | 6 (35.3) | 14 (82.4) | 9 (52.9) | 11 (64.7) | 8 (47.1) | 12 (70.6) | 9 (56.2) | 10 (58.8) | |
p (2-sided) |
0.01
| NS | NS | NS |