Altered Expression of Genes in Signaling Pathways Regulating Proliferation of Hematopoietic Stem and Progenitor Cells in Mice with Subchronic Benzene Exposure
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Animals and Treatments
2.3. Blood Parameters, Relative Organ Weight and Bone Marrow Smears
2.4. Collection of Hematopoietic Stem cells
2.5. Colony Forming Cells Assay
2.6. Real-time PCR Assay
2.7. Statistical Analysis
Primer Name | Primer Set | Sequence (5′-3′) |
---|---|---|
β-actin | Outer | Forward: CGAGCGGTTCCGATGCCCTG; Reverse: ACGCAGCTCAGTAACAGTCCGC |
Inner | Forward: ATCCGCAAAGACCTGT; Reverse: GGGTGTAACGCAACTAAG | |
β-catenin | Outer | Forward: GACACCTCCCAAGTCCTTT; Reverse: CCTCTGAGCCCTAGTCATT |
Inner | Forward: ACACCTCCCAAGTCCTTTATGAAT; Reverse: CCCGTCAATATCAGCTACTTGCT | |
c-Myc | Outer | Forward: GCGCTCTCCTTCCTCGGACTCGC; Reverse: GGTTTGCCTCTTCTCCACAG; |
Inner | Forward: GCCCCTAGTGCTGCATGAG; Reverse: CCACAGACACCACATCAATTTCTT | |
Notch1 | Outer | Forward: GGCCTTGAGACAGGCAACA; Reverse: CTGCACTGGCCTCCAGCAA |
Inner | Forward: CCTTGAGACAGGCAACAGTGAAG; Reverse: GAGAGTATCGGGCAGCCAAGT | |
cyclinD1 | Outer | Forward: GCGCTTCCAACCCACCCTCCATG; Reverse: GCGCCGCAGGCTTGACTCCAGAA |
Inner | Forward: GGGCAGCCCCAACAACTT; Reverse: GCAGTCCGGGTC ACACTTG | |
Bmi1 | Outer | Forward: CAGAGGGATGGACTGACGA; Reverse: GCGTGAGGGAACTGTGGGTGA |
Inner | Forward: GAAAGTGACTCTGGGAGTGACAAG; Reverse: GGGACTGGGCAAACAGGAA | |
p53 | Outer | Forward: CTTCCACCTGGGCTTCCTG; Reverse: GTGGCGCTGACCCACAACT |
Inner | Forward: TGGCGCTGACCCACAACT; Reverse: CCAAGTCTGTTATGTGCACGTACTCT |
3. Results
3.1. Body Weight and Relative Organ Weight
Group | Relative Organ Weight (Mean ± SD) | |||
---|---|---|---|---|
Relative Liver Weight | Relative Spleen Weight | Relative Lung Weight | Relative Kidney Weight | |
Control | 5.31 ± 0.89 | 0.38 ± 0.10 | 0.88 ± 0.44 | 1.93 ± 0.10 |
Benzene 1 | 5.14 ± 0.39 | 0.41 ± 0.10 | 0.91 ± 0.44 | 1.92 ± 0.17 |
Benzene 2 | 5.37 ± 0.57 | 0.52 ± 0.14*,# | 0.98 ± 0.43 | 1.89 ± 0.25 |
3.2. Blood Parameters and Bone Marrow Smears
3.3. Frequency of LSK Cells and Number of Colony Forming
Time Points | Blood Parameters (Mean ± SD) | ||||
---|---|---|---|---|---|
Group | WBC (109/L) | RBC (1012/L) | Hgb (g/L) | Pit (109/L) | |
Three months after benzene exposure | Control | 5.05 ± 1.56 | 8.28 ± 0.32 | 138.42 ± 4.07 | 661.92 ± 87.01 |
Benzene 1 | 1.60 ± 0.72* | 5.63 ± 1.15* | 114.00 ± 19.8* | 374.00 ± 144.18* | |
Benzene 2 | 2.19 ± 1.16* | 5.84 ± 1.39* | 90.00 ± 37.98* | 318.67 ± 163.54* | |
Ten months post-benzene exposure | Control | 7.43 ± 2.03 | 8.23 ± 1.22 | 130.27 ± 22.12 | 514.40 ± 85.60 |
Benzene 1 | 5.81 ± 1.82 | 7.59 ± 1.07 | 117.75 ± 16.12 | 677.37 ± 155.22* | |
Benzene 2 | 6.10 ± 2.49 | 6.75 ± 1.27*,# | 106.65 ± 13.10* | 701.73 ± 196.21* |
3.4. Expression of β-catenin, c-Myc, Notch1, Cyclin D1, Bmi 1 and P53
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sun, R.; Zhang, J.; Xiong, M.; Wei, H.; Tan, K.; Yin, L.; Pu, Y. Altered Expression of Genes in Signaling Pathways Regulating Proliferation of Hematopoietic Stem and Progenitor Cells in Mice with Subchronic Benzene Exposure. Int. J. Environ. Res. Public Health 2015, 12, 9298-9313. https://doi.org/10.3390/ijerph120809298
Sun R, Zhang J, Xiong M, Wei H, Tan K, Yin L, Pu Y. Altered Expression of Genes in Signaling Pathways Regulating Proliferation of Hematopoietic Stem and Progenitor Cells in Mice with Subchronic Benzene Exposure. International Journal of Environmental Research and Public Health. 2015; 12(8):9298-9313. https://doi.org/10.3390/ijerph120809298
Chicago/Turabian StyleSun, Rongli, Juan Zhang, Mengzhen Xiong, Haiyan Wei, Kehong Tan, Lihong Yin, and Yuepu Pu. 2015. "Altered Expression of Genes in Signaling Pathways Regulating Proliferation of Hematopoietic Stem and Progenitor Cells in Mice with Subchronic Benzene Exposure" International Journal of Environmental Research and Public Health 12, no. 8: 9298-9313. https://doi.org/10.3390/ijerph120809298