Allopregnanolone Alters the Gene Expression Profile of Human Glioblastoma Cells
Abstract
:1. Introduction
2. Results
2.1. 3α-THP and F Promote Gene Expression Changes in U87 Cells
2.2. 3α-THP and F Increase the mRNA Levels of Proteins Involved in Several Cellular and Metabolic Processes
2.3. 3α-THP and F Increase the Expression Level of Genes Selected for Validation
2.4. CREB1 and CEBPa Could Mediate 3α-THP-Dependent Transcriptional Effects
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Treatments
4.2. RNA Extraction and Microarrays
4.3. Validation of Selected Differentially Expressed Genes by RT-qPCR
4.4. Bioinformatic Analysis of Transcription Factor Binding Sites
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gene Full Name | Treatment vs. V (Fc) | Treatment vs. V (p-Value) | Ontogeny Categories | ||
---|---|---|---|---|---|
3α-THP | F | 3α-THP | F | ||
ESF1 nucleolar pre-rRNA processing protein homolog (ESF1) | 3.07 | 2.01 | 0.0103 | 0.044 | ND |
Translocated promoter region, nuclear basket protein (TPR) | 2.75 | - | 0.010 | - |
|
Replication timing regulatory factor 1 (RIF1) | 2.69 | - | 0.015 | - | ND |
RAD50 double-strand break repair protein (RAD50) | 2.31 | 2.16 | 0.010 | 0.022 |
|
Rho-associated coiled-coil containing protein kinase 1 (ROCK1) | 1.94 | 2.04 | 0.009 | 0.018 |
|
Rho-associated coiled-coil containing protein kinase 2 (ROCK2) | 1.94 | 1.8 | 0.041 | 0.034 |
|
REV3-like, DNA directed polymerase zeta catalytic subunit (REV3L) | 1.86 | - | 0.014 | - |
|
Pericentriolar material 1 protein (PCM1) | 1.68 | 1.54 | 0.037 | 0.044 |
|
Dynein cytoplasmic 2 heavy chain 1 (DYNC2H1) | 1.64 | 1.55 | 0.001 | 0.023 |
|
Coiled-coil domain containing 91, P56 protein (CCDC91) | 1.63 | - | 0.04 | - | ND |
Gene | Primer Sequence 5′→3′ | Amplified Fragment (bp) |
---|---|---|
ESF1 | FW: GCTCCTCGTGCTGATGAGATTA | 176 |
RV: TGCTCTTCCTTCATCCTCTCCT | ||
PCM1 | FW: TCAAGACAAGAAAAGCGTCTGC | 180 |
RV: GGGCTGAATGTCTGTTCCTACT | ||
TPR | FW: TTTGGCACAGTTTCGGCTAC | 164 |
RV: TCTTCCTCAGTTCCTACAGGTG | ||
RIF1 | FW: TAATAAGGTTCGCCGTGTCTCC | 177 |
RV: CCTTTGGCTGAAGTGGTATTATGC | ||
REV3L | FW: TGAGAAATGAGGTGGCTCTAAC | 168 |
RV: CACGGACACGGCTAACATAA | ||
RAD50 | FW: GCCTCACTCATCATTCGCCT | 168 |
RV: AAGCTGGAAGTTACGCTGCT | ||
ROCK1 | FW: ATGGAACCAGTACAACAAGCTGA | 159 |
RV: GCATCTTCGACACTCTAGGGC | ||
ROCK2 | FW: GAAGAGCAGCAGAAGTGGGT | 170 |
RV: GGCAGTTAGCTAGGTTTGTTTGG | ||
CCDC91 | FW: AAGTCAGGAAACTGTTAAGGCAG | 152 |
RV: ACAGGCTTCTTTGGCGGAT | ||
DYNC2H1 | FW: GCTTGGCGGAGCAGATTAAA | 159 |
RV: CCAGGATGCCCGATTCAGTAT | ||
18S | FW: AGTGAAACTGCAATGGCTC | 167 |
RV: CTGACCGGGTTGGTTTTGAT |
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Zamora-Sánchez, C.J.; Del Moral-Morales, A.; Hernández-Vega, A.M.; Hansberg-Pastor, V.; Salido-Guadarrama, I.; Rodríguez-Dorantes, M.; Camacho-Arroyo, I. Allopregnanolone Alters the Gene Expression Profile of Human Glioblastoma Cells. Int. J. Mol. Sci. 2018, 19, 864. https://doi.org/10.3390/ijms19030864
Zamora-Sánchez CJ, Del Moral-Morales A, Hernández-Vega AM, Hansberg-Pastor V, Salido-Guadarrama I, Rodríguez-Dorantes M, Camacho-Arroyo I. Allopregnanolone Alters the Gene Expression Profile of Human Glioblastoma Cells. International Journal of Molecular Sciences. 2018; 19(3):864. https://doi.org/10.3390/ijms19030864
Chicago/Turabian StyleZamora-Sánchez, Carmen J., Aylin Del Moral-Morales, Ana M. Hernández-Vega, Valeria Hansberg-Pastor, Ivan Salido-Guadarrama, Mauricio Rodríguez-Dorantes, and Ignacio Camacho-Arroyo. 2018. "Allopregnanolone Alters the Gene Expression Profile of Human Glioblastoma Cells" International Journal of Molecular Sciences 19, no. 3: 864. https://doi.org/10.3390/ijms19030864