Gap Junction Intercellular Communication Positively Regulates Cisplatin Toxicity by Inducing DNA Damage through Bystander Signaling
Abstract
:1. Introduction
2. Results
2.1. Cisplatin Treatment at High Density Sensitizes Cancer Cell Lines
2.2. Connexin Expression in Lung and Ovarian Cancer Cells
2.3. Cx43 Knockdown Cells Leads to Cisplatin Resistance at High-Density Treatment
2.4. Cisplatin Treatment Produces DNA DSBs in Bystander Cells
2.5. DSB Production in Bystander Cells Depends on Functional GJIC
2.6. Functional Gap Junction Communication Potentiates Cisplatin Cytotoxicity in DNA Repair Deficient Cells
3. Discussion
4. Materials and Methods
4.1. Colony Survival Assay
4.2. Chemicals
4.3. Cell Culture
4.4. siRNA Sequence and Transfections
4.5. Western Blot
4.6. RNA Isolation, Reverse Transcription and Transcript Abundance
4.7. Lucifer Yellow Dye-Transfer Assay
4.8. Gamma-H2AX Phosphorylation for DNA DSB Measurement
4.9. Immunofluorescence with Cisplatin-DNA Intra-Adduct Specific Antibody
4.10. Mutation Frequency and Survival Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
NSCLC | Non-small cell lung cancer |
GJ | Gap junction |
GJIC | Gap junction intercellular communication |
ICL | Interstrand crosslinks |
DSBs | Double strand breaks |
Cx43 | Connexin-43 |
ERCC1 | Excision repair cross-complementation group 1 |
XPF | Xeroderma pigmentation group F |
siRNA | Small interfering RNA |
StaRT–PCR | Standardized reverse transcription–polymerase chain reaction |
ACTB | β-Actin |
RIBE | Radiation induced bystander effect |
BE | Bystander effect |
SAHA | Suberoylanilide hydroxamic acid |
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Arora, S.; Heyza, J.R.; Chalfin, E.C.; Ruch, R.J.; Patrick, S.M. Gap Junction Intercellular Communication Positively Regulates Cisplatin Toxicity by Inducing DNA Damage through Bystander Signaling. Cancers 2018, 10, 368. https://doi.org/10.3390/cancers10100368
Arora S, Heyza JR, Chalfin EC, Ruch RJ, Patrick SM. Gap Junction Intercellular Communication Positively Regulates Cisplatin Toxicity by Inducing DNA Damage through Bystander Signaling. Cancers. 2018; 10(10):368. https://doi.org/10.3390/cancers10100368
Chicago/Turabian StyleArora, Sanjeevani, Joshua R. Heyza, Elaine C. Chalfin, Randall J. Ruch, and Steve M. Patrick. 2018. "Gap Junction Intercellular Communication Positively Regulates Cisplatin Toxicity by Inducing DNA Damage through Bystander Signaling" Cancers 10, no. 10: 368. https://doi.org/10.3390/cancers10100368