Serum-Derived Exosomal MicroRNA Profiles Can Predict Poor Survival Outcomes in Patients with Extranodal Natural Killer/T-Cell Lymphoma
Abstract
:Simple Summary
Abstract
1. Introduction
2. Methods
2.1. Study Design and Patients
2.2. Isolation of Exosomes from Serum Samples and Nanostring nCounter Analyses
2.3. Cell Lines and Isolation of Exosomes from Cell Culture Medium
2.4. Transmission Electron Microscopy and Nanoparticle Tracking Analysis
2.5. RNA Extraction
2.6. Quantitative Real-Time Polymerase Chain Reaction
2.7. Western Blot
2.8. Establishment of Etoposide-Resistant Cell Lines and Cell Viability Assay
2.9. miRNA Transfection and Cytokine Array
2.10. Transwell Co-Culture Assay
2.11. Data Preparation and Statistical Analyses
3. Results
3.1. Isolation of Exosomes from Patients’ Serum in the Training Cohort
3.2. NKTL Cell Line-Derived Exosomal miRNA Profiles
3.3. Validation of the Prognostic Relevance of Exosomal miRNAs in ENKTL
3.4. Expression of miR-21-5p, miR-4454, and miR-320e in Treatment-Resistant NKTL Cell Lines
3.5. Effect of Upregulated miR-21-5p and miR-320e on Cytokine Production
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Characteristics | Groups | Poor Outcomes n (%) | Favorable Outcomes n (%) | p |
---|---|---|---|---|
Age | ≤60 years | 13 (59) | 20 (87) | 0.047 |
>60 years | 9 (41) | 3 (13) | ||
Sex | Male | 13 (59) | 15 (65) | 0.763 |
Female | 9 (41) | 8 (35) | ||
Performance status | ECOG 0/1 | 15 (68) | 23 (100) | 0.004 |
ECOG ≥ 2 | 7 (32) | 0 | ||
Serum LDH | Normal | 2 (9) | 17 (74) | <0.001 |
Increased | 20 (91) | 6 (26) | ||
Stage | I/II | 1 (5) | 23 (100) | <0.001 |
III/IV | 21 (95) | 0 | ||
Extranodal involvement | Number 0/1 | 4 (18) | 20 (87) | <0.001 |
Number ≥ 2 | 18 (82) | 3 (13) | ||
Bone marrow | Not involved | 8 (36) | 23 (100) | <0.001 |
Involved | 14 (64) | 0 | ||
Blood EBV DNA | Not detected | 0 | 23 (100) | <0.001 |
Detected | 22 (100) | 0 | ||
PINK-E risk | Low | 0 | 23 (100) | <0.001 |
Intermediate | 4 (18) | 0 | ||
High | 18 (82) | 0 | ||
Primary treatment | SMILE | 11 (50) | 2 (9) | <0.001 |
VIDL | 5 (23) | 0 | ||
VIPD | 3 (14) | 0 | ||
MIDLE | 1 (5) | 0 | ||
CCRT followed by VIDL | 10 (44) | |||
CCRT followed by VIPD | 5 (22) | |||
CCRT followed by MIDLE | 3 (13) | |||
Other | 1 (5) | 3 (13) | ||
Relapse or progression | Did not occur | 0 | 17 (74) | <0.001 |
Occurred | 22 (100) | 6 (26) | ||
Survival outcome | Alive | 0 | 21 (91) | <0.001 |
Dead | 22 (100) | 2 (9) |
Characteristics | miR-4454 | miR-21-5p | miR-320e | ||||||
---|---|---|---|---|---|---|---|---|---|
Low n (%) | High n (%) | p | Low n (%) | High n (%) | p | Low n (%) | High n (%) | p | |
Age | |||||||||
≤60 years | 27 (64) | 35 (81) | 0.091 | 30 (71) | 32 (74) | 0.810 | 29 (69) | 33 (77) | 0.471 |
>60 years | 15 (36) | 8 (19) | 12 (29) | 11 (26) | 13 (31) | 10 (23) | |||
Sex | |||||||||
Male | 32 (76) | 27 (63) | 0.240 | 30 (71) | 29 (67) | 0.815 | 27 (64) | 32 (74) | 0.353 |
Female | 10 (24) | 16 (37) | 12 (29) | 14 (33) | 15 (36) | 11 (26) | |||
Serum LDH | |||||||||
Normal | 28 (67) | 19 (44) | 0.050 | 30 (71) | 17 (40) | 0.004 | 25 (60) | 22 (51) | 0.515 |
Increased | 14 (33) | 24 (56) | 12 (29) | 26 (60) | 17 (40) | 21 (49) | |||
Stage | |||||||||
I/II | 29 (69) | 21 (49) | 0.078 | 33 (79) | 17 (40) | <0.001 | 28 (67) | 22 (51) | 0.188 |
III/IV | 13 (31) | 22 (51) | 9 (21) | 26 (60) | 14 (33) | 21 (49) | |||
Extranodal involvement | |||||||||
Number 0/1 | 26 (62) | 22 (51) | 0.384 | 29 (69) | 19 (44) | 0.029 | 27 (64) | 21 (49) | 0.191 |
Number ≥ 2 | 16 (38) | 21 (49) | 13 (31) | 24 (56) | 15 (36) | 22 (51) | |||
Bone marrow | |||||||||
Not involved | 39 (93) | 35 (81) | 0.195 | 39 (93) | 35 (81) | 0.195 | 39 (93) | 35 (81) | 0.195 |
Involved | 3 (7) | 8 (19) | 3 (7) | 8 (19) | 3 (7) | 8 (19) | |||
Blood EBV DNA | |||||||||
Not detected | 20 (48) | 13 (30) | 0.122 | 24 (57) | 9 (21) | 0.001 | 16 (38) | 17 (40) | >0.999 |
Detected | 22 (52) | 30 (70) | 18 (43) | 34 (79) | 26 (62) | 26 (60) | |||
PINK-E risk | |||||||||
Low | 22 (52) | 18 (41) | 0.420 | 26 (62) | 14 (33) | 0.021 | 21 (50) | 19 (44) | 0.146 |
Intermediate | 9 (21) | 8 (19) | 7 (17) | 10 (23) | 11 (26) | 6 (14) | |||
High | 11 (26) | 17 (40) | 9 (21) | 19 (44) | 10 (24) | 18 (42) | |||
Relapse or progression | |||||||||
Did not occur | 25 (60) | 16 (37) | 0.052 | 27 (64) | 14 (33) | 0.005 | 21 (50) | 20 (47) | 0.829 |
Occurred | 17 (40) | 27 (63) | 15 (36) | 29 (67) | 21 (50) | 23 (53) | |||
Survival | |||||||||
Alive | 30 (71) | 22 (51) | 0.075 | 32 (76) | 20 (47) | 0.007 | 29 (69) | 23 (54) | 0.183 |
Dead | 12 (29) | 21 (49) | 10 (24) | 23 (53) | 13 (31) | 20 (46) |
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Ryu, K.J.; Lee, J.Y.; Choi, M.E.; Yoon, S.E.; Cho, J.; Ko, Y.H.; Shim, J.H.; Kim, W.S.; Park, C.; Kim, S.J. Serum-Derived Exosomal MicroRNA Profiles Can Predict Poor Survival Outcomes in Patients with Extranodal Natural Killer/T-Cell Lymphoma. Cancers 2020, 12, 3548. https://doi.org/10.3390/cancers12123548
Ryu KJ, Lee JY, Choi ME, Yoon SE, Cho J, Ko YH, Shim JH, Kim WS, Park C, Kim SJ. Serum-Derived Exosomal MicroRNA Profiles Can Predict Poor Survival Outcomes in Patients with Extranodal Natural Killer/T-Cell Lymphoma. Cancers. 2020; 12(12):3548. https://doi.org/10.3390/cancers12123548
Chicago/Turabian StyleRyu, Kyung Ju, Ji Young Lee, Myung Eun Choi, Sang Eun Yoon, Junhun Cho, Young Hyeh Ko, Joon Ho Shim, Won Seog Kim, Chaehwa Park, and Seok Jin Kim. 2020. "Serum-Derived Exosomal MicroRNA Profiles Can Predict Poor Survival Outcomes in Patients with Extranodal Natural Killer/T-Cell Lymphoma" Cancers 12, no. 12: 3548. https://doi.org/10.3390/cancers12123548