NEDD9 Inhibition by miR-25-5p Activation Is Critically Involved in Co-Treatment of Melatonin- and Pterostilbene-Induced Apoptosis in Colorectal Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. MLT+Ptero Exerted Synergistic Cytotoxicity in CRCs
2.2. MLT+Ptero Cleaved PARP and Caspase-3 and Modulated NEDD9-Related Proteins in CRCs
2.3. MLT+Ptero Increased the Number of TUNEL-Positive Cells and ROS Production and Suppressed Ki67-Positive Cells in CRCs
2.4. Differentially Expressed microRNA Profile in MLT- and/or Ptero-Treated HT29 Cells
2.5. Late-Stage Autophagy Inhibitor CQ, but not 3-MA, Enhanced Cytotoxicity and Decreased p62 and Activated LC3II in Atorvastatin-Treated H596, H460, and H1299 Cells
2.6. Inhibition of miR-25-5p Rescued the Decreased Expression of NEDD in MLT+Ptero-Treated HT29 Cells
2.7. Inhibition of miR-25-5p Reduced the Antiproliferative and Apoptotic Effects of MLT+Ptero in CRCs
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Chemicals and Reagents
4.3. MTT Assay
4.4. Colony Formation Assay
4.5. Observation of Apoptotic Morphological Features
4.6. TUNEL Assay
4.7. ROS Measurement
4.8. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
4.9. Transfection Assay
4.10. Immunofluorescence Assay
4.11. Western Blotting
4.12. MicroRNA Microarray and Data Analysis
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Jung, J.H.; Shin, E.A.; Kim, J.-H.; Sim, D.Y.; Lee, H.; Park, J.E.; Lee, H.-J.; Kim, S.-H. NEDD9 Inhibition by miR-25-5p Activation Is Critically Involved in Co-Treatment of Melatonin- and Pterostilbene-Induced Apoptosis in Colorectal Cancer Cells. Cancers 2019, 11, 1684. https://doi.org/10.3390/cancers11111684
Jung JH, Shin EA, Kim J-H, Sim DY, Lee H, Park JE, Lee H-J, Kim S-H. NEDD9 Inhibition by miR-25-5p Activation Is Critically Involved in Co-Treatment of Melatonin- and Pterostilbene-Induced Apoptosis in Colorectal Cancer Cells. Cancers. 2019; 11(11):1684. https://doi.org/10.3390/cancers11111684
Chicago/Turabian StyleJung, Ji Hoon, Eun Ah Shin, Ju-Ha Kim, Deok Yong Sim, Hyemin Lee, Ji Eon Park, Hyo-Jung Lee, and Sung-Hoon Kim. 2019. "NEDD9 Inhibition by miR-25-5p Activation Is Critically Involved in Co-Treatment of Melatonin- and Pterostilbene-Induced Apoptosis in Colorectal Cancer Cells" Cancers 11, no. 11: 1684. https://doi.org/10.3390/cancers11111684