Expression of Growth Hormone-Releasing Hormone and Its Receptor Splice Variants in a Cohort of Hungarian Pediatric Patients with Hematological and Oncological Disorders: A Pilot Study
Abstract
:1. Introduction
2. Results
2.1. PCR Analyses of the Expression of GHRH and SV1 of GHRH-Rs
2.2. Western Blot Analysis
2.3. GHRH Receptor Binding Studies
3. Discussion
4. Materials and Methods
4.1. Sample Collecting
4.2. RNA Isolation and Reverse Transcription Reaction (RT-PCR)
4.3. Reverse Transcription PCR (RT-PCR)
4.4. RT-PCR Reaction for GHRH and SV1 of GHRH-R
4.5. Western Blot Analysis of GHRH-R Protein
4.6. Preparation of Membranes and Radioligand Binding Studies
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Models Investigated | |
---|---|
Breast | Renal |
Ovarian | Osteosarcoma |
Endometrial | Ewing sarcoma |
Prostate | Glioblastoma |
Lung | Esophageal squamous cell carcinoma |
Pancreatic | Pleural mesothelioma |
Colorectal | Acute myeloid leukemia |
Sample | Clinical Condition | GHRH mRNA | SV1 mRNA |
---|---|---|---|
1 | HL | + | + |
2 | RMS | + | + |
3 | RMS | + | + |
4 | TR/adenocarcinoma components | + | + |
5 | ALL | + | + |
6 | ALL | + | + |
7 | ALL | + | + |
8 | ALL | − | - |
9 | ALL | + | + |
10 | BMH | + | + |
11 | FD | + | + |
12 | JM | + | + |
13 | CBN | − | − |
14 | HS | − | − |
15 | ITP | − | − |
Sample | Clinical Condition | GHRH-R Protein | GHRH-R Binding | |
---|---|---|---|---|
Kd (nM) | Bmax (fmol/mg Prot.) | |||
1 | HL | + | 1.37 | 398.7 |
2 | RMS | + | 3.84 | 222.1 |
3 | RMS | + | 4.38 | 245.0 |
4 | TR/adenocarcinoma components | + | 8.99 | 475.0 |
5 | ALL | N/A | N/A | |
6 | ALL | N/A | N/A | |
7 | ALL | N/A | N/A | |
8 | ALL | N/A | N/A | |
9 | ALL | N/A | N/A | |
10 | BMH | + | 7.27 | 733.0 |
11 | FD | + | 4.81 | 261.4 |
12 | JM | + | 1.35 | 294.6 |
13 | CBN | N/A | N/A | |
14 | HS | N/A | N/A | |
15 | ITP | N/A | N/A |
Sample | Clinical Condition | Sex | Sample Origin | Age at Sampling | Disease Outcome |
---|---|---|---|---|---|
1 | HL | F | Lymph node | 15 years | Alive |
2 | RMS | M | Pelvic tumor tissue | 12.5 years | † Died |
3 | RMS | F | Neck mass biopsy | 8 years | † Died |
4 | TR/adenocarcinoma components | M | Mediastinal tumor tissue | 14.5 years | † Died |
5 | ALL | F | Bone marrow | 5 years | Alive |
6 | ALL | M | Bone marrow | 13 years | Alive |
7 | ALL | M | Bone marrow | 9.5 years | † Died |
8 | ALL | M | Bone marrow | 6.5 years | Alive |
9 | ALL | M | Peripheral blood | 15 years | † Died |
10 | BMH | M | Mediastinal mass | 4 years | Alive |
11 | FD | F | Bone tumor tissue | 10.5 years | Alive |
12 | JM | M | Right pelvic bone tumor tissue | 2 years | Alive |
13 | CBN | M | Bone marrow | 12.5 months | Alive |
14 | HS | F | Bone marrow | 9 months | Alive |
15 | ITP | M | Bone marrow | 10 months | Alive |
Primers | Forward Sequences | Reverse Sequences | Tm (°C) Annealing Temperature | Product Size (bp) |
---|---|---|---|---|
ACTB | 5′-GGCATCCTCACCCCCTGAAGTA-3′ | 5′-GGGGTGAAGGTCTCAAA-3′ | 60 °C | 203 bp |
GHRH | 5′-GGAGTTGTGGCTAGAGAG-3′ | 5′-TGTCTGTCTACCTGACGACCAA-3′ | 64 °C | 150 bp |
SV1 | 5′-GGAGTTGTGGCTAGAGAG-3′ | 5′-GCATAGAACAGTGGAGAAG-3′ | 67 °C | 373 bp |
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Juhász, É.; Szabó, Z.; Schally, A.V.; Király, J.; Fodor, P.; Kónya, G.; Dezső, B.; Szabó, E.; Halmos, G.; Kiss, C. Expression of Growth Hormone-Releasing Hormone and Its Receptor Splice Variants in a Cohort of Hungarian Pediatric Patients with Hematological and Oncological Disorders: A Pilot Study. Int. J. Mol. Sci. 2024, 25, 8831. https://doi.org/10.3390/ijms25168831
Juhász É, Szabó Z, Schally AV, Király J, Fodor P, Kónya G, Dezső B, Szabó E, Halmos G, Kiss C. Expression of Growth Hormone-Releasing Hormone and Its Receptor Splice Variants in a Cohort of Hungarian Pediatric Patients with Hematological and Oncological Disorders: A Pilot Study. International Journal of Molecular Sciences. 2024; 25(16):8831. https://doi.org/10.3390/ijms25168831
Chicago/Turabian StyleJuhász, Éva, Zsuzsanna Szabó, Andrew V. Schally, József Király, Petra Fodor, Gábor Kónya, Balázs Dezső, Erzsébet Szabó, Gábor Halmos, and Csongor Kiss. 2024. "Expression of Growth Hormone-Releasing Hormone and Its Receptor Splice Variants in a Cohort of Hungarian Pediatric Patients with Hematological and Oncological Disorders: A Pilot Study" International Journal of Molecular Sciences 25, no. 16: 8831. https://doi.org/10.3390/ijms25168831