Mechanism of Interaction between hsa_circ_0002854 and MAPK1 Protein in PM2.5-Induced Apoptosis of Human Bronchial Epithelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell and Cell Culture
2.2. Collection and Treatment of Particulate Matter
2.3. Cell Counting Kit-8 Assay
2.4. Flow Cytometry Experiment
2.5. RT-qPCR Analysis
2.6. RNase R Digestive Experiment
2.7. Nucleocytoplasmic Separation Experiment
2.8. Fluorescence In Situ Hybridization Experiment (FISH)
2.9. Small Interfering RNA (siRNA) Transfection
2.10. RNA Pull-Down Experiment
2.11. Protein Extraction and Western Blot Analysis
2.12. Statistical Analysis
3. Result
3.1. Survival Rate of 16HBE Cells after 24 h of Exposure to PM2.5
3.2. Apoptosis Rate of 16HBE Cells after 24 h of Exposure to PM2.5
3.3. qRT-PCR-Validated circRNAs Associated with Apoptosis Pathways
3.4. RNase R Digestive Test
3.5. Nucleocytoplasmic Separation Experiment
3.6. Fluorescence In Situ Hybridization Experiment
3.7. Interference Efficiency of siRNA
3.8. hsa_circ_0002854 Functional Verification Experiment
3.9. Study on the Mechanism of hsa_circ_0002854
3.10. Western Blot Assay Verified the Regulatory Effect of hsa_circ_0002854 on MAPK1 Protein
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer | Primer Sequence (5′ to 3′) |
---|---|
ACTB-F | TCTGGCGGCACCACCATGTAC |
ACTB-R | CTGCTTGCTGATCCACATCTGCTG |
U6_F1 | CTCGCTTCGGCAGCACA |
U6_R1 | AACGCTTCACGAATTTGCGT |
hsa_circ_0002854_F | GTCAGCTCAAACCACTAGTTATACAGGTAC |
hsa_circ_0002854_R | CAGTCATCGTCATCATCATTTTCGTCC |
hsa_circ_0003036_F | GGAGAAATTAACTGAATGCCAAGGG |
hsa_circ_0003036_R | TGATTTCGTATGATCCGGTCGC |
hsa_circ_0034296_F | TTCCAGCCAGCCTGCTAAAC |
hsa_circ_0034296_R | AGCTGTTTCCTCCATTGCTGT |
hsa_circ_0000780_F | TTAGCACACTGGTGGAAGATCA |
hsa_circ_0000780_R | CCAAGATACAACCAAAAGCCAT |
hsa_circ_0003352_F | ACTGTTGGATGCAGATGGATGAG |
hsa_circ_0003352_R | GAAATAGCCCAGGGGAAGGAGAT |
hsa_circ_0001571_F | CGTGCTGAAAGCCGAGGG |
hsa_circ_0001571_R | CCAAAGCCTCCGCTGTCC |
hsa_circ_0005791_F | AGAACAGTGAACAAGGTGG |
hsa_circ_0005791_R | AAACAGTTAATATCATCCC |
hsa_circ_0071106_F | GGTTTTGTTTTGACATAGAAGCTGCTG |
hsa_circ_0071106_R | CTGCCTCTTGTAAATGTGAGTCTTTCT |
Probe Name | Probe Sequence | |
---|---|---|
full-length sequence | sense | GTACGAGTGGCTGCTTTACAGAATCTGGTGAAGATAAT GTCCTTATATTATCAGTACATGGAGACATATATGGGTCC TGCTCTTTTTGCAATCACAATCGAAGCAATGAAAAGTGA CATTGATGAGGTGGCTTTACAAGGGATAGAATTCTGGTC CAATGTCTGTGATGAGGAAATGGATTTGGCCATTGAAGC TTCAGAGGCAGCAGAACAAGGACGGCCCCCTGAGCACAC CAGCAAGTTTTATGCGAAGGGAGCACTACAGTATCTGGT TCCAATCCTCACACAGACACTAACTAAACAGGACGAAAA TGATGATGACGATGACTGGAACCCCTGCAAAGCAGCAGG GGTGTGCCTCATGCTTCTGGCCACCTGCTGTGAAGATGA CATTGTCCCACATGTCCTCCCCTTCATTAAAGAACACAT CAAGAACCCAGATTGGCGGTACCGGGATGCAGCAGTGAT GGCTTTTGGTTGTATCTTGGAAGGACCAGAGCCCAGTCA GCTCAAACCACTAGTTATACAG |
anti-sense | CTGTATAACTAGTGGTTTGAGCTGACTGGGCTCTGGTC CTTCCAAGATACAACCAAAAGCCATCACTGCTGCATCC CGGTACCGCCAATCTGGGTTCTTGATGTGTTCTTTAAT GAAGGGGAGGACATGTGGGACAATGTCATCTTCACAGC AGGTGGCCAGAAGCATGAGGCACACCCCTGCTGCTTTG CAGGGGTTCCAGTCATCGTCATCATCATTTTCGTCCTG TTTAGTTAGTGTCTGTGTGAGGATTGGAACCAGATACT GTAGTGCTCCCTTCGCATAAAACTTGCTGGTGTGCTCA GGGGGCCGTCCTTGTTCTGCTGCCTCTGAAGCTTCAAT GGCCAAATCCATTTCCTCATCACAGACATTGGACCAGA ATTCTATCCCTTGTAAAGCCACCTCATCAATGTCACTTT TCATTGCTTCGATTGTGATTGCAAAAAGAGCAGGACCC ATATATGTCTCCATGTACTGATAATATAAGGACATTAT CTTCACCAGATTCTGTAAAGCAGCCACTCGTAC | |
joint sequence | sense | CCACTAGTTATACAGGTACGAGTGGCTGCT |
anti-sense | AGCAGCCACTCGTACCTGTATAACTAGTGG |
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Hong, J.; Tan, Y.; Wang, Y.; Wang, H.; Li, C.; Jin, W.; Wu, Y.; Ni, D.; Peng, X. Mechanism of Interaction between hsa_circ_0002854 and MAPK1 Protein in PM2.5-Induced Apoptosis of Human Bronchial Epithelial Cells. Toxics 2023, 11, 906. https://doi.org/10.3390/toxics11110906
Hong J, Tan Y, Wang Y, Wang H, Li C, Jin W, Wu Y, Ni D, Peng X. Mechanism of Interaction between hsa_circ_0002854 and MAPK1 Protein in PM2.5-Induced Apoptosis of Human Bronchial Epithelial Cells. Toxics. 2023; 11(11):906. https://doi.org/10.3390/toxics11110906
Chicago/Turabian StyleHong, Jinchang, Yi Tan, Yuyu Wang, Hongjie Wang, Caixia Li, Wenjia Jin, Yi Wu, Dechun Ni, and Xiaowu Peng. 2023. "Mechanism of Interaction between hsa_circ_0002854 and MAPK1 Protein in PM2.5-Induced Apoptosis of Human Bronchial Epithelial Cells" Toxics 11, no. 11: 906. https://doi.org/10.3390/toxics11110906