The Comparative Experimental Study of Sodium and Magnesium Dichloroacetate Effects on Pediatric PBT24 and SF8628 Cell Glioblastoma Tumors Using a Chicken Embryo Chorioallantoic Membrane Model and on Cells In Vitro
Abstract
:1. Introduction
2. Results
2.1. Stereomicroscopic Findings of Transplanted PBT24 and SF8628 Cell Tumors on CAM
2.2. PBT24 and SF8628 Tumor Growth, Tumor Invasion into CAM Rate, the Number of Blood Vessels, and CAM Thickness
2.3. The PCNA Expression of PBT24 and SF8628 in Control and DCA-Treated Tumors
2.4. EZH2 Expression of PBT24 and SF8628 in Control and DCA-Treated Tumors
2.5. The p53 Expression in Control and DCA-Treated PBT24 and SF8628 Tumors
2.6. The Survivin Expression in the Control and DCA-Treated PBT24 and SF8628 Tumors
2.7. The Expression of NKCC1, KCC2, SLC5A8, E- and N-Cadherins Genes in the Studied PBT24 and SF8628 Cells Groups
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Cell Culture
4.2. Groups Studied for PBT24 and SF8628 Tumors on CAM
4.3. Application of the CAM Model to the Study of PBT24 and SF8626 Tumors on CAM
4.4. Immunohistochemical Study
4.5. Extraction of RNA from Study Cells and Determination of the Gene Expression in PBT24 and SF8628 Cells
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Study Group | n | Invasion Frequency (%) | Number of Blood Vessels | CAM Thickness (µm) |
---|---|---|---|---|
Median (Range) | ||||
PBT24-control | 13 | 76.9 | 15 (6–28) | 300.9 (65.2–700.9) |
PBT24-5 mM NaDCA | 13 | 53.9 a,b | 12 (5–29) | 236.02 (31.3–484.4) |
PBT24-10 mM NaDCA | 12 | 16.7 c | 11.5 (3–25) | 199.2 (54.4–627.6) |
PBT24-2.5 mM MgDCA | 11 | 45.5 | 9 (2–23) | 138.6 (21.3–383.0) n |
PBT24-5 mM MgDCA | 11 | 18.2 d | 7 (2–27) i | 96.5 (36.3–591.9) o |
SF8628-control | 13 | 84.6 | 15 (5–21) | 282.5 (47.85–539.7) |
SF8628-5 mM NaDCA | 14 | 28.6 e | 6 (1–22) j | 247.5 (86.76–879.3) |
SF8628-10 mM NaDCA | 13 | 7.7 f | 5 (2–16) k | 199.4 (52.09–776.2) |
SF8628-2.5 mM MgDCA | 14 | 57.1 g | 10.5 (5–23) l | 244.2 (55.02–409.1) |
SF8628-5 mM MgDCA | 11 | 27.3 h | 8 (2–31) m | 140.5 (50.56–636.4) |
Study Group | PBT24 Tumor PCNA-Positive Cells (%) | SF8628 Tumor PCNA-Positive Cells (%) | ||
---|---|---|---|---|
n | Median (Range) | n | Median (Range) | |
Control | 9 | 63.8 (34.9–81.0) | 8 | 90.8 (76.3–100) |
5 mM NaDCA | 6 | 15.0 (10.0–31.1) a,b | 7 | 31.5 (29.0–48.5) g,h |
10 mM NaDCA | 8 | 10.5 (2.4–24.3) c,d | 7 | 19.1 (9.1–24.6) i |
2.5 mM MgDCA | 7 | 43.1 (19.5–93.6) | 6 | 52.2 (19.3–60.7) j,k,l |
5 mM MgDCA | 6 | 20.7 (16.4–31.7) e,f | 7 | 16.7 (8.2–44.6) m |
Study Group | PBT24 Tumor EZH2-Positive Cells (%) | SF8628 Tumor EZH2-Positive Cells (%) | ||
---|---|---|---|---|
n | Median (Range) | n | Median (Range) | |
Control | 6 | 71.0 (42.6–78.7) | 8 | 85.4 (72.0–97.5) |
5 mM NaDCA | 6 | 18.8 (9.8–78.7) a,b | 6 | 41.6 (18.0–53.5) g,h |
10 mM NaDCA | 7 | 19.6 (14.9–44.6) c,d | 7 | 25.9 (13.8–35.3) i |
2.5 mM MgDCA | 7 | 44.7 (21.5–94.8) | 6 | 38.4 (9.9–61.1) j |
5 mM MgDCA | 7 | 17.9 (2.8–44.4) e,f | 6 | 22.4 (8.9–50.0) k |
Study Group | PBT24 Tumor p53-Positive Cells (%) | SF8628 Tumor p53-Positive Cells (%) | ||
---|---|---|---|---|
n | Median (Range) | n | Median (Range) | |
Control | 7 | 75.1 (61.2–96.9) | 8 | 85.3 (56.7–92.9) |
5 mM NaDCA | 8 | 53.7 (33.9–75.5) a | 6 | 40.7 (27.1–48.9) h |
10 mM NaDCA | 8 | 29.6 (11.5–52.3) b,c,d | 6 | 19.5 (11.3–52.6) i |
2.5 mM MgDCA | 7 | 51.9 (32.0–91.0) e | 6 | 56.3 (27.8–87.1) j |
5 mM MgDCA | 6 | 24.2 (19.3–61.6) f,g | 7 | 44.9 (5.4–52.8) k |
Study Group | PBT24 Tumor Survivin- Positive Cells (%) | SF8628 Tumor Survivin-Positive Cells (%) | ||
---|---|---|---|---|
n | Median (Range) | n | Median (Range) | |
Control | 6 | 53.8 (40.6–81.1) | 6 | 52.8 (45.7–89.6) |
5 mM NaDCA | 6 | 33.1 (4.3–56.8) | 7 | 45.0 (15.6–62.3) |
10 mM NaDCA | 8 | 8.5 (1.3–29.3) a,b,c | 7 | 12.4 (8.1–37.0) d,e |
2.5 mM MgDCA | 7 | 43.9 (8.4–58.7) | 6 | 37.0 (13.0–8.6) |
5 mM MgDCA | 6 | 19.6 (0.9–79.4) | 6 | 22.4 (10.6–58.1) f |
Study Group | n | CT Mean | ΔCT Mean ± SD | ΔΔCT | |
---|---|---|---|---|---|
SLC12A2 | GAPDH | ||||
PBT24-control | 6 | 22.95 | 19.37 | 3.58 ± 0.73 | |
PBT24-3 mM NaDCA | 6 | 22.72 | 18.69 | 4.03 ± 0.48 | 0.45 |
PBT24-1.5 mM MgDCA | 6 | 23.00 | 19.31 | 3.69 ± 1.33 | 0.11 |
SF8628-control | 6 | 22.89 | 19.02 | 3.88 ± 0.21 | |
SF8628-3 mM NaDCA | 6 | 23.61 | 19.77 | 3.84 ± 0.29 | −0.04 |
SF8628-1.5 mM MgDCA | 6 | 23.66 | 19.63 | 4.04 ± 0.27 | 0.16 |
SLC12A5 | GAPDH | ΔCT Mean ± SD | ΔΔCT | ||
PBT24-control | 6 | 32.56 | 19.37 | 13.19 ± 0.83 | |
PBT24-3 mM NaDCA | 6 | 31.94 | 18.69 | 13.25 ± 0.72 | 0.06 |
PBT24-1.5 mM MgDCA | 6 | 32.39 | 19.31 | 13.08 ± 1.19 | −0.11 |
SF8628-control | 6 | 36.83 | 19.02 | 17.81 ± 0.43 a | |
SF8628-3 mM NaDCA | 6 | 37.84 | 19.77 | 18.07 ± 0.81 b | 0.260 |
SF8628-1.5 mM MgDCA | 6 | 37.66 | 19.63 | 18.04 ± 0.29 c | 0.223 |
SLC5A8 | GAPDH | ΔCT Mean ± SD | ΔΔCT | ||
PBT24-control | 6 | 39.42 | 19.37 | 20.04 ± 1.82 | |
PBT24-3 mM NaDCA | 6 | 38.91 | 18.69 | 20.22 ± 1.81 | 0.18 |
PBT24-1.5 mM MgDCA | 6 | 39.47 | 19.31 | 20.17 ± 2.24 | 0.12 |
SF8628-control | 6 | 38.97 | 19.02 | 19.96 ± 0.59 | |
SF8628-3 mM NaDCA | 6 | 40.06 | 19.77 | 20.29 ± 0.68 | 0.34 |
SF8628-1.5 mM MgDCA | 6 | 40.03 | 19.63 | 20.41 ± 0.54 | 0.45 |
Study Group | n | CT Mean | ΔCT Mean ± SD | ΔΔCT | |
---|---|---|---|---|---|
CDH1 | GAPDH | ||||
PBT24-control | 6 | 33.48 | 19.37 | 14.104 ± 1.1 | |
PBT24-3 mM NaDCA | 6 | 33.61 | 18.69 | 14.92 ± 1.24 | 0.82 |
PBT24-1.5 mM MgDCA | 6 | 33.95 | 19.31 | 14.65 ± 1.56 | 0.54 |
SF8628-control | 6 | 38.69 | 19.02 | 19.67 ± 0.51 a | |
SF8628-3 mM NaDCA | 6 | 38.59 | 19.77 | 18.83 ± 0.79 b | −0.84 |
SF8628-1.5 mM MgDCA | 6 | 37.36 | 19.63 | 17.73 ± 0.32 c,d,e | −1.94 |
CDH2 | GAPDH | ΔCT Mean ± SD | ΔΔCT | ||
PBT24-control | 6 | 22.92 | 19.37 | 3.55 ± 0.98 | |
PBT24-3 mM NaDCA | 6 | 22.55 | 18.69 | 3.86 ± 0.85 | 0.31 |
PBT24-1.5 mM MgDCA | 6 | 22.28 | 19.31 | 2.98 ± 1.09 | −0.57 |
SF8628-control | 6 | 23.45 | 19.02 | 4.43 ± 0.23 | |
SF8628-3 mM NaDCA | 6 | 23.95 | 19.77 | 4.18 ± 0.39 | −0.25 |
SF8628-1.5 mM MgDCA | 6 | 24.14 | 19.63 | 4.52 ± 0.19 f | 0.08 |
Control and Treated Group | Invasion, No. of Vessels, CAM Thickness | PCNA | EZH2 | p53 | Survivin | |||||
---|---|---|---|---|---|---|---|---|---|---|
PBT24 n | SF8628 n | PBT24 n | SF8628 n | PBT24 n | SF8628 n | PBT24 n | SF8628 n | PBT24 n | SF8628 n | |
Control | 13 | 13 | 9 | 8 | 6 | 8 | 7 | 8 | 6 | 6 |
5 mM NaDCA | 13 | 14 | 6 | 7 | 6 | 6 | 8 | 6 | 6 | 7 |
10 mM NaDCA | 12 | 13 | 8 | 7 | 7 | 7 | 8 | 6 | 8 | 7 |
2.5 mM MgDCA | 11 | 14 | 7 | 6 | 7 | 6 | 7 | 6 | 7 | 6 |
5 mM MgDCA | 11 | 11 | 6 | 7 | 7 | 6 | 6 | 7 | 6 | 6 |
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Damanskienė, E.; Balnytė, I.; Valančiūtė, A.; Lesauskaitė, V.; Alonso, M.M.; Stakišaitis, D. The Comparative Experimental Study of Sodium and Magnesium Dichloroacetate Effects on Pediatric PBT24 and SF8628 Cell Glioblastoma Tumors Using a Chicken Embryo Chorioallantoic Membrane Model and on Cells In Vitro. Int. J. Mol. Sci. 2022, 23, 10455. https://doi.org/10.3390/ijms231810455
Damanskienė E, Balnytė I, Valančiūtė A, Lesauskaitė V, Alonso MM, Stakišaitis D. The Comparative Experimental Study of Sodium and Magnesium Dichloroacetate Effects on Pediatric PBT24 and SF8628 Cell Glioblastoma Tumors Using a Chicken Embryo Chorioallantoic Membrane Model and on Cells In Vitro. International Journal of Molecular Sciences. 2022; 23(18):10455. https://doi.org/10.3390/ijms231810455
Chicago/Turabian StyleDamanskienė, Eligija, Ingrida Balnytė, Angelija Valančiūtė, Vaiva Lesauskaitė, Marta Marija Alonso, and Donatas Stakišaitis. 2022. "The Comparative Experimental Study of Sodium and Magnesium Dichloroacetate Effects on Pediatric PBT24 and SF8628 Cell Glioblastoma Tumors Using a Chicken Embryo Chorioallantoic Membrane Model and on Cells In Vitro" International Journal of Molecular Sciences 23, no. 18: 10455. https://doi.org/10.3390/ijms231810455