Tumor Cell-Derived Microvesicles Induced Not Epithelial-Mesenchymal Transition but Apoptosis in Human Proximal Tubular (HK-2) Cells: Implications for Renal Impairment in Multiple Myeloma
Abstract
:1. Introduction
2. Results
2.1. Validation and Characteristics of Myeloma Cell-Derived Microvesicles ( MM-MVs)
2.2. MM-MVs Inhibit Viability and Induce Apoptosis in Human Kidney-2 Cells (HK-2 Cells)
2.3. MM-MVs Activate Apoptic Pathways of Caspase-3, -8, -9 and Bcl-2 Family Members
2.4. MM-MVs Up-Regulate E-Cadherin Protein and Down-Regulate Vimentin Protein in HK-2 Cells
2.5. Selective miRNAs in MM-MVs that Confer Caspase-3-Induced Apoptosis in HK-2 Cells
2.6. CD138+ Circulating MV (cirMV) Counts Positively Correlate with Renal Impairment
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. MV Isolation and Quantification
4.3. Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM)
4.4. Small RNA Sequencing and Data Analyses
4.5. Cell Viability Detection
4.6. Apoptosis Detection
4.7. Western Blotting
4.8. Immunofluorescent Stainning
4.9. Construction of miRNA Regulatory Network
4.10. Realtime PCR
4.11. Clinical Samples Collection
4.12. Quantitative Detection of cirMVs
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
RI | Renal Impairment |
MM | Multiple Myeloma |
MVs | Microvesicles |
EMT | Epithelial-Mesenchymal Transition |
RPM | Reads Per Million Mapped Reads |
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Clinical Characteristics | Patient Number (n) |
---|---|
Total | 61 |
Sex | |
Male | 40 |
Female | 21 |
Median age, years (range) | 59 (40–82) |
Durie-Salmon stage | |
I | 3 |
II | 11 |
III | 38 |
International staging system | |
I | 6 |
II | 25 |
III | 27 |
Type of monoclonal Ig | |
IgG | 27 |
IgA | 16 |
IgD | 3 |
LC a only | 8 |
Renal function | |
Group 1 (SCr b < 2 mg/dL) | 45 |
Group 2 (SCr ≥ 2 mg/dL) | 16 |
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Share and Cite
Zhao, A.; Kong, F.; Liu, C.-J.; Yan, G.; Gao, F.; Guo, H.; Guo, A.-Y.; Chen, Z.; Li, Q. Tumor Cell-Derived Microvesicles Induced Not Epithelial-Mesenchymal Transition but Apoptosis in Human Proximal Tubular (HK-2) Cells: Implications for Renal Impairment in Multiple Myeloma. Int. J. Mol. Sci. 2017, 18, 513. https://doi.org/10.3390/ijms18030513
Zhao A, Kong F, Liu C-J, Yan G, Gao F, Guo H, Guo A-Y, Chen Z, Li Q. Tumor Cell-Derived Microvesicles Induced Not Epithelial-Mesenchymal Transition but Apoptosis in Human Proximal Tubular (HK-2) Cells: Implications for Renal Impairment in Multiple Myeloma. International Journal of Molecular Sciences. 2017; 18(3):513. https://doi.org/10.3390/ijms18030513
Chicago/Turabian StyleZhao, Aiqi, Fancong Kong, Chun-Jie Liu, Guoxin Yan, Fei Gao, Hao Guo, An-Yuan Guo, Zhichao Chen, and Qiubai Li. 2017. "Tumor Cell-Derived Microvesicles Induced Not Epithelial-Mesenchymal Transition but Apoptosis in Human Proximal Tubular (HK-2) Cells: Implications for Renal Impairment in Multiple Myeloma" International Journal of Molecular Sciences 18, no. 3: 513. https://doi.org/10.3390/ijms18030513