“…The mean spectra generated from each of the cell population was overlaid in all the cell lines (Figure 1). Mean spectra showed key spectral features -proteins (~1005 cm -1 , Amide-III ~1260 cm -1 , Amide-I ~1664 cm -1 ), lipids (~1305 cm -1 , ~1742 cm -1 , ~1445 cm -1 CH2 bending, DNA (~720 cm -1 , ~1096 cm -1 , ~1340 cm -1 ) and cytochrome C (~750 cm -1 , ~1128 cm -1 , ~1583 cm -1 ) [17][18][19][20] . Mean Raman spectrum of HeLa showed high intensity bands for lipids (~1305 cm -1 , ~1742 cm -1 ) , Amide-III (~1260 cm -1 ), cytochrome C (~750 cm -1 , ~1583 cm -1 ) in resistant cells.…”
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.
“…The mean spectra generated from each of the cell population was overlaid in all the cell lines (Figure 1). Mean spectra showed key spectral features -proteins (~1005 cm -1 , Amide-III ~1260 cm -1 , Amide-I ~1664 cm -1 ), lipids (~1305 cm -1 , ~1742 cm -1 , ~1445 cm -1 CH2 bending, DNA (~720 cm -1 , ~1096 cm -1 , ~1340 cm -1 ) and cytochrome C (~750 cm -1 , ~1128 cm -1 , ~1583 cm -1 ) [17][18][19][20] . Mean Raman spectrum of HeLa showed high intensity bands for lipids (~1305 cm -1 , ~1742 cm -1 ) , Amide-III (~1260 cm -1 ), cytochrome C (~750 cm -1 , ~1583 cm -1 ) in resistant cells.…”
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.
“…However, our ML framework has not yet included calculations on any possible chemical reactions upon LTP irradiation. Nevertheless, the prospect of adjusting the dose rate of LTP by tuning the process parameters offers great potential for a wide range of applications in medical treatments 32 , 33 . Figure 3 Dose-rate dependence with frequency and voltage.…”
Low-temperature plasmas have quickly emerged as alternative and unconventional types of radiation that offer great promise for various clinical modalities. As with other types of radiation, the therapeutic efficacy and safety of low-temperature plasmas are ubiquitous concerns, and assessing their dose rates is crucial in clinical settings. Unfortunately, assessing the dose rates by standard dosimetric techniques has been challenging. To overcome this difficulty, we proposed a dose-rate assessment framework that combined the predictive modeling of plasma-induced damage in DNA by machine learning with existing radiation dose-DNA damage correlations. Our results indicated that low-temperature plasmas have a remarkably high dose rate that can be tuned by various process parameters. This attribute is beneficial for inducing radiobiological effects in a more controllable manner.
“…A recent study by Abramczyk et al showed a correlation between the intensity of cytochrome Raman bands (750 and 1585 cm –1 ) and the different grades of breast and brain cancer tissue and cell lines . Our earlier studies have shown increased intensity of cytochrome bands in short-term radiation-exposed MCF-7 and gingivobuccal carcinoma-derived ITOC-03 cells. , Corroborating our earlier findings, here we report a similar increase in cytochrome Raman bands at 1585 and 750 cm –1 in radioresistant MCF-7 cells compared to that in parental MCF-7 cells. Studies have shown that radiation-induced ROS levels cause DNA damage, mitochondrial outer membrane permeabilization, the release of mitochondrial cytochrome C into the cytoplasm, activation of caspases, and eventually apoptosis .…”
Recapitulating radioresistant
cell features in pertinent cell line
models is essential for deciphering fundamental cellular mechanisms.
The limited understanding of passage and cell cycle phases on radioresistant
cells revived post-cryopreservation led us to investigate the effect
of sub-culturing in parental and radioresistant MCF-7 cells. In this
study, the radioresistant cells showed high-intensity nucleic acid
and cytochrome bands, which are potentially a radiation-induced spectral
marker. Raman spectroscopy data showed dynamic biochemical alterations
in revived radioresistant G2/M synchronized cells at early
cell passages 1 and 3 with stabilization at a latter cell passage,
5. The study highlights the importance of cell passaging and cell
cycle phases in potentially changing the biochemical parameters during
in vitro experiments after the revival of radioresistant cells post-cryopreservation.
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