Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Breast Tumor Samples
2.2. Raman Spectroscopy
2.3. Multivariate Analysis
2.4. Breast Tumor Sample Staining for Confocal Microscopy Imaging
2.5. In Vitro Synthesis of DNA-HAp Particles
3. Results
3.1. Molecular Raman Imaging and Advanced Multivariate Analysis of Tissue Calcifications
3.2. Study of DNA Mineralization in HAp Microcalcifications
3.2.1. Reference Raman Spectral Databases and Literature
3.2.2. Identification of DNA in HAp Calcifications Found in Breast Cancer Tissues
3.2.3. Fluorescence Staining of Microcalcifications with DAPI
3.3. Raman Images Reveal the Presence and Distribution of Biomolecules Inside HAp Calcifications
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Diagnosis | Oxalate | HAp | DNA | Size a | Shape b | Localisation c | |
---|---|---|---|---|---|---|---|
1 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, D = 20 µm | S | t |
2 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, D = 20 µm | R | t |
3 | Infiltrating ductal carcinoma (IDC) | No | Yes | No | m, D = 10 µm | S | t |
3 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m | R | t |
4 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, D = 20 µm | R | d |
5 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, D = 20 µm | S | t |
5 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 30 × 20 µm | R | t |
6 | Infiltrating ductal carcinoma (IDC) | No | No | No | - | - | - |
7 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, D = 10 µm | R | t |
8 | Infiltrating ductal carcinoma (IDC) | No | No | No | - | - | - |
9 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 25 × 15 µm | R | t |
9 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 20 µm | S | t |
10 | Infiltrating ductal carcinoma (IDC), multifocal | No | Yes | Yes | m, 30 µm | R | t |
11 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m | R | t |
12 | Infiltrating ductal carcinoma (IDC) | No | No | - | - | - | - |
13 | Infiltrating ductal carcinoma (IDC) | No | Yes | No | b | R | t |
14 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes (nc) | S | R | t |
15 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | b | R | t |
16 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 100 µm | R | d |
17 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 40 × 50 µm | R | t |
18 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | s, D = 7 µm | S | t |
19 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m | R | t |
20 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m | R | d |
21 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m | R | d |
22 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | m, 20 × 10 µm | R | t |
23 | Infiltrating ductal carcinoma (IDC) | No | Yes | Yes | b | R | t |
24 | Infiltrating lobular carcinoma (ILC) | No | Yes | Yes | m, D = 20 µm | R | t |
25 | Infiltrating lobular carcinoma (ILC) | Yes | No | No | s | S | t |
25 | Infiltrating lobular carcinoma (ILC) | No | Yes | Yes | m | R | t |
26 | Infiltrating lobular carcinoma (ILC) | No | Yes | Yes | m | R | t |
Component 2: DNA | |||
---|---|---|---|
Band/cm−1 | Assignment | Band/cm−1 | Assignment |
1679 | T, DNA base | 1084 | Phosphodiester groups in nucleic acids |
1656 | T, G, C (ring breathing modes of the DNA/RNA bases); amide I (protein); C=C lipids | 1060 | PO2-stretching of DNA |
1605 | Cytosine (NH2) | 1004 | Phenylalanine |
1551 | Guanine | 972 | T, ribose, Phosphate monoester groups of phosphorylated proteins and cellular nucleic acids |
1457 | A, C, T DNA bases | 937 | G, A DNA bases |
1452 | CH2 CH3 deformation | 855 | Phosphate group |
1426 | Deoxyribose (B, Z marker) | 824 | O-P-O stretch DNA |
1342 | A, G DNA bases | 785 | U, T, C, O-P-O |
1322 | G, DNA base | 754 | T, DNA |
1297 | C, palmitic acid, CH2 | 721 | A, DNA base |
1271 | C, G DNA bases | 645 | G DNA base |
1261 | A, G, C DNA bases | 625 | A, T DNA bases |
1205 | A, T, ring breathing mode of DNA bases | 597 | C DNA base |
1131 | A DNA base | 543 | C DNA base |
1104 | O-P-O backbone stretch DNA, U | 423 | T, U DNA bases |
Component 3: Lipid | Component 4: Polysaccharides and Proteins | Component 5: Cytochrome C | |||
---|---|---|---|---|---|
Band/cm−1 | Assignment | Band/cm−1 | Assignment | Band/cm−1 | Assignment |
436 | Cholesterol, cholesterol ester | 424 | Polysaccharide | 749 | Cytochrome C |
597 | Phosphatidylinositol | 496 | Glycogen | 1002 | Phenylalanine |
624 | Cholesterol ester | 850 | Polysaccharide | 1128 | Cytochrome C |
721 | Phosphatidylcholine, Sphingomyelin | 1002 | Phenylalanine | 1174 | CH tyrosine phenylalanine |
1081 | Phosphatidylcholine, membrane lipids | 1062 | C-C carbohydrates | 1232 | Amide III |
1133 | Membrane lipids (PC, PE, SM) | 1126 | Disaccharides, glucose, sucrose, C-O stretching carbohydrates | 1312 | Cytochrome C |
1300 | Membrane lipids (PC, PE) | 1205 | Amide III proteins | 1339 | tryptophan |
1314 | CH3CH2 twisting of lipid | 1270 | Amide III proteins | 1449 | C-H proteins |
1450 | CH (lipids and proteins) | 1297 | CH2 deformation | 1587 | Cytochrome C |
1653 | C=C lipids, proteins | 1322 | CH deformation proteins | 1658 | Amide I proteins |
1342 | CH deformation protein and carbohydrates | ||||
1447 | CH2 deformation proteins | ||||
1553 | Tryptophan, Amide II | ||||
1603 | Phenylalanine | ||||
1656 | Amide I proteins |
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Marro, M.; Rodríguez-Rivero, A.M.; Araujo-Andrade, C.; Fernández-Figueras, M.T.; Pérez-Roca, L.; Castellà, E.; Navinés, J.; Mariscal, A.; Julián, J.F.; Turon, P.; et al. Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging. Cancers 2021, 13, 2658. https://doi.org/10.3390/cancers13112658
Marro M, Rodríguez-Rivero AM, Araujo-Andrade C, Fernández-Figueras MT, Pérez-Roca L, Castellà E, Navinés J, Mariscal A, Julián JF, Turon P, et al. Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging. Cancers. 2021; 13(11):2658. https://doi.org/10.3390/cancers13112658
Chicago/Turabian StyleMarro, Monica, Anna M. Rodríguez-Rivero, Cuauhtémoc Araujo-Andrade, Maria Teresa Fernández-Figueras, Laia Pérez-Roca, Eva Castellà, Jordi Navinés, Antonio Mariscal, Joan Francesc Julián, Pau Turon, and et al. 2021. "Unravelling the Encapsulation of DNA and Other Biomolecules in HAp Microcalcifications of Human Breast Cancer Tissues by Raman Imaging" Cancers 13, no. 11: 2658. https://doi.org/10.3390/cancers13112658