Mr. Parikshit J. Patel Profile

Parikshit J. Patel

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Proceedings Article | 11 August 2023 Paper

Raman spectroscopy in in vitro cell cultures: understanding chemoresistance

Parikshit Patel, Abhiram Natu, Arti Hole, Sanjay Gupta, C. Murali Krishna

Proceedings Volume 12629, 126290C (2023) https://doi.org/10.1117/12.2670317

KEYWORDS: Raman spectroscopy, Cancer, Liver cancer, Resistance, Oncology, Cervical cancer, Principal component analysis, Proteins, Cancer detection, Radiotherapy

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Over the past several decades, research and innovations in the field of cancer therapy have garnered significant attention globally. Despite these advancements, incidences of recurrence and relapse have been a major obstacle in cancer treatment. Moreover, the underlined cause of death due to cancer is largely due to delayed detection and therapy resistance. It is evident that early detection and prediction of therapy resistance can benefit in designing personalized treatment regimen and better prognosis. Therefore, vibrational spectroscopic techniques such as Raman spectroscopy (RS) are being explored. RS is a rapid, inexpensive and objective method that extracts useful biochemical information. It provides a global molecular fingerprint of the cellular milieu, highlighting the physiological state of the system. In this study, we have evaluated RS to characterize and delineate chemoresistant and parental cell types of cervical and liver cancer cell lines HeLa, SiHa, HepG2 and Hep3B respectively. Doxorubicin-induced resistant cell lines were developed using incremental drug dosing method to obtain a desirable resistance index value. Raman spectra of these cell lines were recorded to understand biochemical profiles and stratify resistant and parental groups using confocal Raman microscope (WITec alpha 300R) and were preprocessed. Mean spectra were computed and were subjected to multivariate analyses such as Principal Component Analysis (PCA) and Principal Component based Linear Discriminant Analysis (PC-LDA). Key spectral features such as protein, DNA, lipid and cytochrome levels were observed higher in the resistant cell populations than that in corresponding parental cell populations. PC-LDA results showed good classification between parental and resistant cell populations for all the cell lines. Thus, RS can be prospectively explored in clinical settings for better stratification of cancer patients, enabling effective therapeutic outcomes.

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