Raman Spectroscopy Combined With Statistics Showed Differential Expression Of Neuroblastoma And Related Tumor Components

Objectives:Each organism consists of lipids,amino acids,nucleic acids,carbohydrates,etc.When the biological tissue cells proliferate,attenuate,mutate,or appeare lesions change,the composition and conformation of the biomolecules accompanying will change.These changes can be Raman scattering.It is detected that a unique Raman scattering spectrum is formed,and information on biomolecules can be obtained by studying the spectrum.At present,invasive pathological examination of tissue is still used as a diagnostic gold standard,and no effective rapid and non-invasive diagnosis method has been found in the clinic.To investigate the subtle differences of molecules in neuroblastoma,ganglioneuroma and ganglioneuroblastoma and in different types of neuroblastoma,We analyzed their Raman spectra.To proved that neuroblastoma,ganglioneuroma and ganglioneuroblastoma,and their related classifications have different Raman fingerprints,which may become the basis for distinguishing the pathological types and related classifications.To find molecules with different characteristics in different classifications of Raman fingerprints,which may become the key to study the mechanism of action of related molecules in the development of tumors.Methods:Colleccting Surgical or biopsy specimens of 29 case of neuroblastoma,14case of ganglioneuroblastoma and ganglioneuroma that have been paraffin-embedded cryopreserved in the pathology department of Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology for nearly ten years,and all disease information of related specimens used as a diagnostic gold standardand as well as classification of tumors in children.Each specimen was thawed,sliced into 10?m,fixed on a gold-plated slide,and dewaxed.According to the existing disease information,the corresponding specimens were divided into groups of neuroblastoma?29 cases?and ganglioneuroblastoma/ganglioneuroma?14 cases?.The corresponding specimens were divided into groups of FH?19 cases?and UFH?19cases?according to clinical prognosis classification;divided into groups of T1?20cases?,T2?7cases?,T3-4?15cases?according to International Neuroblastoma Staging System;divided into groups of death and recurrence?12 cases?,1 year survival?20 cases?,and 3 years survival?5 cases?according to the actual survival.Neuroblastoma was divided into groups of undifferentiated?6 cases?and differentiation?19 cases?according to the degree of tissue differentiation;divided into groups of low-risk?22 cases?and middle-high risk group?19 cases?according to the degree of risk.we collected at least 24 spectra from different regions of each sample using a Renishaw Raman microscope with 532-nm laser,selecting the spectral area as400-2000 cm^-1with 3 accumulations of 10 seconds each.After the initial processing of removing noise,Raw spectra were analyzed using principle component analysis and discriminant function analysis?PCA-LDA?by MATLAB.Results:The one-dimensional and two-dimensional graphs of PCA-LDA showed that thediscriminationbetweengroupsofneuroblastomaand ganglioneuroblastoma/ganglioneuroma were significant?P<0.05?,the covariance matrix analysis between the above groups showed that there were significant differences in the cross peaks in the bio-fingerprint region,same as the other groups.Vector cluster analysis showed that there was a statistically significant difference in the molecular absorbance between groups of neuroblastoma and ganglioneuroblastoma/ganglioneuroma,as well as other groups.In the neuroblastoma,the molecular absorbance of Tyrosine(1169 cm^-1),DNA/RNA bases(1373 cm^-1),CH bending band of lipids(1439 cm^-1)was significantly increased?P<0.001?,in carbohydrates(905 cm^-1),amide III(1325 cm^-1)molecular absorbance was significantly reduced?P<0.001?.Conclusions:Thebiologicalinformationofneuroblastomaand ganglioneuroblastoma/ganglioneuroma can be reflected from the Raman spectrum.Raman can provide molecular spectral information with different characteristics between different tumors and different types of tumor.The results indicate that the biomolecular composition and structure between neuroblastoma and non-neuroblastoma in the study are significantly different,with different molecular fingerprints.There are different Raman fingerprints in different clinical stages of this type of tumor,also in different prognostic classifications,different risk levels,different degrees of tumor tissue differentiation,and in different actual prognosis.These fingerprints may become the basis for distinguishing the pathological types of such tumors and related classifications,and have the potential to diagnose and classify.Molecules such as tyrosine,carbohydrates,and carotenoids,which have significant differences in Raman fingerprints,may become the key to study the molecular mechanism of related molecules in the development of such tumors.