Short-Chain Fatty Acids Suppress mTOR Signaling in Colon Cancer Cells via Long Non-Coding RNA RMST
Abstract
:1. Introduction
2. Results
2.1. SCFAs Induced RMST Expression in Colon Cancer Cells
2.2. RMST Is Required for SCFA to Reduce mTOR Activation
2.3. RMST Suppresses mTOR Activation
2.4. RMST Modulates SCFAs-Induced Autophagy
2.5. TSC2 Is Involved in the Suppression of mTOR Signaling by SCFA/RMST
2.6. Genomic Editing of RMST Gene Affects mTOR Activities and TSC2 Expression
3. Discussion
4. Materials and Methods
4.1. Constructs and Gene Cloning
4.2. Cell Culture and Stable Transfection
4.3. siRNA Transfections
4.4. RNA Isolation and Real-Time RT-PCR
4.5. RMST Genomic Deletion with CRISPR/CAS9 System
4.6. Western Blot
4.7. ELISA for mTOR pser2448
4.8. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target 1 | TGAAACTTGGAGCAAAGTGG(AGG) |
Target 2 | GCTGAATATCTTCAGGAAAA(TGG) |
Target 3 | TTTTTATTTTGCAGATTCTC(AGG) |
Target 4 | CTATATTTTGTTAGAAAGGA(AGG) |
Target 5 | ACTTAGCATTCCATCCGTGC(AGG) |
Target 6 | CTTATTCTGAACTTTCAGAC(TGG) |
Target 7 | AACCTCTATTGTATTCCAGT(TGG) |
Target 8 | TGTCTGGCTTTCCTTGCAGT(GGG) |
Target 9 | (CCT)GCTCGATGTTTGCATTTAAA |
RMST | ||
siRNA1 | sense | CGGAGACUUGGUAGUGAAAUUCUTC |
antisense | GAAGAAUUUCACUACCAAGUCUCCGUG | |
siRNA2 | sense | GUUUGAUUGACCUUUGUUAGGGUGA |
antisense | UCACCCUAACAAAGGUCAAUCAAACUU | |
siRNA3 | sense | CGAAUUAUUUGCAAUGCCAUUUAAT |
antisense | AUUAAAUGGCAUUGCAAAUAAUUCGUU |
Name | Forward Primer | Reverse Primer |
---|---|---|
qPCR | CCCAGTGCCACCAACTGGGAT | CCAGCTGGCGGGTACGATCAG |
gRMST1 | GGG AAA CAG AGG GAG CAC | GGT ATG GAA CAA GGC AGC |
gRMST2 | ATC AAT GCC CCT ACT CAC | ACG CTG CTC TGC ATT TTC |
gRMST3 | TAG CCC GGC TTT TAG GTC | ACC CGT CAG TTT CCA AGA |
gRMST4 | AGC TCT GTC GTT CCA CCT | GTC ACT CCT CGG AAA GC |
gRMST5 | CAG TAG TGC TTG GGA CTC | ACA GCT GAT GTC CCG TTT |
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Wang, J.; Guo, Y.; Fang, X.; Zhang, Y.; Nie, D. Short-Chain Fatty Acids Suppress mTOR Signaling in Colon Cancer Cells via Long Non-Coding RNA RMST. Kinases Phosphatases 2024, 2, 136-150. https://0-doi-org.brum.beds.ac.uk/10.3390/kinasesphosphatases2020008
Wang J, Guo Y, Fang X, Zhang Y, Nie D. Short-Chain Fatty Acids Suppress mTOR Signaling in Colon Cancer Cells via Long Non-Coding RNA RMST. Kinases and Phosphatases. 2024; 2(2):136-150. https://0-doi-org.brum.beds.ac.uk/10.3390/kinasesphosphatases2020008
Chicago/Turabian StyleWang, Jiuhui, Yande Guo, Xiangwei Fang, Yuanqin Zhang, and Daotai Nie. 2024. "Short-Chain Fatty Acids Suppress mTOR Signaling in Colon Cancer Cells via Long Non-Coding RNA RMST" Kinases and Phosphatases 2, no. 2: 136-150. https://0-doi-org.brum.beds.ac.uk/10.3390/kinasesphosphatases2020008