Abstract P4-01-20: Explore the differences and relationships between normal, ADH tissues and DCIS, IDC in breast by shell-isolated nanoparticle-enhanced Raman spectroscopy (SHINERS)

Abstract

Abstract Background: The mechanism that ADH changes to DCIS and even IDC is still not clear now. Raman spectroscopy is a powerful, non-invasive analytical tool which can provide detailed and meaningful information about biochemical composition of tissues at molecular level. Our aim was to find the differences and relationships between normal, ADH tissues and DCIS, IDC in breast based on their various biochemical characteristics by Raman spectroscopy. Methods: The frozen sections were collected from 39 patients (all female; ages 27-74) who underwent surgical resection or mammotome biopsy at the Department of Breast Surgery, the First Hospital of Jilin University. After operation the samples were immediately frozen at -20∼-25°C and two contiguous sections (6 μm thickness) were cut from a sample by freezing microtome. One was stained with haematoxylin and eosin for routine histopathological analysis; the other was detected by Microscopic confocal Raman spectrometer (HORIBA JY Lab800, 633nm) with its mirror image (the H&E section). After the spectra we needed had been obtained, the Au@SiO2 shell-isolated nanoparticles (SHINs) were added to the surface of frozen sections immediately and then the spectra with SHINs were collected. Results: A total of 475 Raman spectra and 470 SHINERS spectra were obtained from 9 normal breast tissues, 7 ADH, 8 DCIS, and 15 IDC breast tissues. The Raman signals were significantly enhanced by SHINs. The main spectral features of normal tissues at around 1301, 1442, 1654, and 1743 cm−1 are attributed to lipids. Meanwhile, no strong protein peaks emerged. The peaks of ADH, DCIS and IDC tissues located at 1004, 1033, 1610 and 1658 cm−1 from vibrational modes of proteins shows more stronger. 830, 880, 970, 1090 and 1157cm−1, representing DNA feature peaks, hold the same rules. Table 1: Peak assignments of the Raman spectra of Breast tissues (wavenumbers in cm-1)NBADHDCISIDCMajor Assignments 880878880O-P-O stretch, DNA1003100410041003phenylalanine1092109210911090O-P-O stretch, DNA1301,14421302,14421303,14431303,1442CH2 twist and CH2 deformation, lipids 15261527C = C stretch of carotene 165816581658Amide I, α-helix1743174717451747C = O stretch, lipids These show that diseased tissues contained more proteins and nucleic acids. The spectra of DCIS and IDC tissues contained 1527 cm−1, stemming from C = C stretching of carotene. The spectral features between ADH, DCIS and IDC tissues are described as followings: firstly, the concentrations of proteins, nucleic acids and carotene increased gradually but lipids content reduced from ADH to IDC. Secondly, ADH and DCIS tissues have extraordinary strong band of amide I, but in IDC tissues the peak almost cannot observed. Additionally, DCIS tissues have the red shifted and broad peak of CH2, which explains acyl backbone of lipids and proteins has fractured. Conclusion: The changes including content and molecular structure of lipids, carotene, proteins and nucleic acids accompany with deterioration, indicating the Raman spectroscopy can be used to explore the differences and relationships between normal, ADH tissues and DCIS, IDC in breast. Simultaneously, it might be used to discover the mechanism that ADH convert to DCIS. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-20.