Substrate Type and Concentration Differently Affect Colon Cancer Cells Ultrastructural Morphology, EMT Markers, and Matrix Degrading Enzymes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Cultures
2.2. RNA Isolation and Real-Time qPCR Analysis
2.3. E-Cadherin Expression by Western Blot
2.4. Statistical Analysis
2.5. Scanning Electron Microscopy
3. Results
3.1. Evaluation of EMT Markers and Matrix Degrading Enzymes in CRC Cells Cultured in Different Matrix Substrates
3.2. Ultrastructural Morphological Features of CRC Cells Cultured on Millipore/Matrigel Mimicking a Normal and a Thick Basement Membrane
3.3. Ultrastructural Morphological Features of LoVo-S and LoVo-R Cells Cultured on Millipore/Collagen Mimicking a Normal and a Desmoplastic Lamina Propria
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Gene | Primer Sequence |
---|---|
E-Cadherin | F: TTCTGCTGCTCTTGCTGTTT, R: TGGCTCAAGTCAAAGTCCTG; |
Vimentin (VIM) | F: AAAACACCCTGCAATCTTTCAGA, R: CACTTTGCGTTCAAGGTCAAGAC; |
SNAIL | F: AGTTTACCTTCCAGCAGCCCTAC, R: AGCCTTTCCCACTGTCCTCATC; |
MMP-2 | F: TGCATCCAGACTTCCTCAGGC, R: TCCTGGCAATCCCTTTGTATGTT; |
MMP-9 | F: GGTGATTGACGACGCCTTTG, R-CTGTACACGCGAGTGAAGGT; |
MMP-14 | F: TGCCATGCAGAAGTTTTACGG, R: TCCTTCGAACATTGGCCTTG; |
Heparanase (HPSE) | F: ATTTGAATGGACGGACTGC R: GTTTCTCCTAACCAGACCTTC; |
GAPDH | F: ACACCCACTCCTCCACCTTT R: TCCACCACCCTGTTGCTGTA; |
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Franchi, M.; Karamanos, K.-A.; Cappadone, C.; Calonghi, N.; Greco, N.; Franchi, L.; Onisto, M.; Masola, V. Substrate Type and Concentration Differently Affect Colon Cancer Cells Ultrastructural Morphology, EMT Markers, and Matrix Degrading Enzymes. Biomolecules 2022, 12, 1786. https://doi.org/10.3390/biom12121786
Franchi M, Karamanos K-A, Cappadone C, Calonghi N, Greco N, Franchi L, Onisto M, Masola V. Substrate Type and Concentration Differently Affect Colon Cancer Cells Ultrastructural Morphology, EMT Markers, and Matrix Degrading Enzymes. Biomolecules. 2022; 12(12):1786. https://doi.org/10.3390/biom12121786
Chicago/Turabian StyleFranchi, Marco, Konstantinos-Athanasios Karamanos, Concettina Cappadone, Natalia Calonghi, Nicola Greco, Leonardo Franchi, Maurizio Onisto, and Valentina Masola. 2022. "Substrate Type and Concentration Differently Affect Colon Cancer Cells Ultrastructural Morphology, EMT Markers, and Matrix Degrading Enzymes" Biomolecules 12, no. 12: 1786. https://doi.org/10.3390/biom12121786