Research ArticleExperimental Studies
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4-Cholesten-3-one Modified the Lipidome of MDA-MB-231 Cells and Potentiated the Effect of Docetaxel

SRISTY SAHA, MIKAËL CROYAL, JEAN-MICHEL HUVELIN and HASSAN NAZIH
Anticancer Research August 2024, 44 (8) 3277-3285; DOI: https://doi.org/10.21873/anticanres.17146

Abstract

Background/Aim: Lipids are essential for energy production, signaling, and membrane formation, hence increased lipid metabolism may lead to cancer growth. 4-cholesten-3-one (4Cone), a sterol metabolite, has various biological activities, including the inhibition of cancer growth. This study examined whether 4Cone could change the lipid profile of triple-negative breast cancer cells (MDA-MB-231) and whether in combination with the anti-cancer chemotherapy docetaxel (TXT) could further reduce cancer aggressiveness. Materials and Methods: The effect of 4Cone, TXT, or their combination (4Cone/TXT) on migration and proliferation was examined utilizing the wound healing and MTT assays. The expression of the lipogenesis-related enzymes was assessed using RT-qPCR and lipid profile was examined using mass spectrometry. Results: 4Cone and TXT individually reduced cell viability and migration of MDA-MB-231 cancer cells; however, their combination (4Cone/TXT) had a greater impact on both attributes. All treated cells showed markedly decreased levels of the multidrug resistance enzyme PGP as well as the lipogenic enzymes FASN, ACC1, SCD1, HMGCR, and DGAT. Furthermore, lipid fingerprints were markedly different in treated cells compared with the untreated group. 4Cone increased the percentage of sphingomyelin (SM) while it decreased the percentage of ceramide (Cer); 4Cone in conjunction with TXT had the reverse effect. Triglyceride levels were reduced in 4Cone- and 4Cone/TXT-treated cells, but interestingly, they increased in TXT-treated cells. Additionally, treated cancer cells exhibited changes in glycerophospholipid subclasses. Conclusion: 4Cone alone or in combination with TXT alters the lipid profile by reducing a key lipogenic enzyme, resulting in the inhibition of cell proliferation and migration.

Key Words:

According to the World Health Organization (WHO), in 2020, about 2.3 million female were diagnosed with breast cancer and 685,000 women died from the disease (1). Breast cancer has always been a global issue due to the yearly increase in diagnoses and death from the disease. Among all the subtype of breast cancer, triple-negative breast cancer (TNBC) is the leading cause of cancer-related women death (2, 3). One of the primary management strategies for TNBC control is chemotherapy (4). Despite the improvement of surgical technique and the development of adjuvant chemotherapeutic regimens, TNBC has a poor survival rate due to chemo resistance and side effects. Docetaxel (TXT) has been considered an excellent chemotherapy for the significant survival benefit in cancer patients, but it’s clinical utility is compromised when primary and acquired resistance are encountered (5, 6). Thus, investigation of novel therapeutic strategies is important for improving survival rate in TNBC.

Lipids, which comprise fatty acids, cholesterol, sphingolipids, and glycerophospholipid are required for membrane architecture, energy storage, and the building blocks of signaling molecules (7-10). To maintain cell proliferation, cancer cells increase the expression of lipogenic enzymes, which changes their phenotype and re-shapes the cell microenvironment (11, 12). The primary process in lipid metabolism is the production of de novo fatty acids (13), which begins with the carboxylation of acetyl-CoA to malonyl-CoA by acetyl-CoA carboxylase (ACC) and continues with the production of palmitic acid by fatty acid synthase (FASN), which is further processed by stearoyl-CoA desaturase 1 (SCD1) to generate monounsaturated fatty acids. Another important lipid metabolic activity is the mevalonate pathway, which involves cholesterol synthesis via 3-hydroxy-3-methyl-glutarylcoenzyme reductases (HMGCR) (14). Furthermore, triglycerides, which are regarded to be the principal energy reserve for cells, are synthesized by the diacylglycerol acyltransferase (DGAT) enzymes (15). Because of the relevance in cancer cell growth and proliferation, multiple studies show that targeting these lipogenesis-related enzymes could be a useful cancer treatment strategy (16, 17).

4-cholesten-3-one (4Cone) is a sterol metabolite, naturally derived from the oxidation of cholesterol in the gastrointestinal tract. In general, 4Cone can be used for a variety of biological functions, including inhibition of cancer cell growth, suppression of metastases, inhibition of beta transgender signaling (18-20). Recent evidence shows that 4Cone has anti-cancer effects on MCF-7 and MDA-MB-231 breast cancer cell lines (21). Based on the previous findings, the aims of this study were to investigate whether 4Cone can modulate the lipidome of the MDA-MB-231 TNBC cell line through the alteration of enzymes involved in lipid synthesis pathways and to determine whether 4Cone alone or in combination with TXT could improve the therapeutic potential of MDA-MB-231 breast cancer cells.

Materials and Methods

Materials. Human TNBC cell line MDAMB-231 was purchased from the European Collection of Animal Cell Cultures (ECACC) (Salisbury, UK). 4-cholesten-3- one, 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), fetal bovine serum (FBS), Dulbecco’s Modified Eagle’s Medium (DMEM), ethanol, penicillin-streptomycin, trypsin, glutamine, dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), primers for qPCR, and triton X-100 were purchased from Sigma Aldrich (Saint-Quentin Fallavier, France). M-PER™ Mammalian Protein Extraction Reagent, TRIzol reagent for RNA isolation, iScriptTM Reverse Transcription Supermix for RTqPCR, and iQ™ SYBR Green Supermix were purchased from Bio-Rad (Marnes-la-Coquette, France). All solvents used for mass spectrometry analysis (ultrapure water, isopropanol, acetonitrile, formic acid and ammonium acetate) were purchased from Biosolve (Valkenswaard, the Netherlands).

Cell line culture. TNBC cells (MDA-MB-231) were grown in DMEM supplemented with 10% FBS, 1% glutamine and 1% penicillin-streptomycin and incubated at 37°C in a humidified atmosphere containing 5% CO2.

Preparation of treatments. 4Cone and TXT were dissolved in absolute ethanol at 5 mM and 1 mM respectively. For the treatment, the stock solution was diluted with a serum-free medium (DMEM, supplemented with 0.1% BSA (Dulbecco’s Modified Eagle’s Medium, 1% penicillin-streptomycin and 1% glutamine).

Cell viability test. MDA-MB-231 cells were spread in a 96-well plate at a density of 10,000 cells/well and grown for 24 h. The cells were treated for 24 h and 48 h at the final concentrations of 25 μM of 4Cone (Ethanol), 100 nM of TXT (Ethanol) and their combination. After recommended times of treatments, 50 μl of MTT solution (2.5 mg/ml in water) was added to each well and incubated for 4 h at 37°C. DMSO (200 μl) was then added to each well to solubilize the formazan product and the absorbance was measured at 540 nm by Spectra-Max 190 microplate reader (Molecular Devices, San Jose, CA, USA).

Cell migration test. MDA-MB-231 cells were spread in a 6-well plate at a density of 5×105 cells/well and grown for 24 h. using a sterile 200 μl pipette tip, a linear scratch wound was made in the center of the cells once the cells had reached maximum confluence. After that, the culture medium was replaced with new medium that had 0.2% ethanol as a control or 25 μM of 4Cone as treatment. The width of the scratched area was measured using an Olympus Inverted Phase Contrast Microscope (Axiovert 40C, Göttingham, Germany) with a 10X phase objective after 0, 24, and 48 h of growth.

RNA extraction and qRT-PCR. MDA-MB-231 cells were spread in a 6-well plate at a density of 5 × 105 cells/well and allowed to adhere overnight. After reaching the maximum confluence, cells were treated with 25 μM of 4Cone, 100 nM of TXT and their combination for 24 h at 37°C. Total RNA was extracted from cultured cells using TriZol Reagent, following the manufacturer’s instructions and the concentration of extracted RNA was measured considering the 260/280 nm absorbance using NanoDrop ND-1000 spectrophotometer (Bio-Rad). Then, 1 μg of total RNA was reverse transcribed into complementary DNA using iScript Reverse Transcription Supermix by following the manufacturer’s protocol (Bio-Rad). An initial priming step of 5 min at 25 °C was followed by a reserve transcription of 30 min at 42°C and a reverse transcription inactivation step of 5 min at 85°C. After cDNA synthesis, quantitative PCR was performed on a MyiQ2 Real-Time PCR Detection System (Bio-Rad) using an iQ™ SYBR Green Supermix (Bio-Rad). The cycling conditions were 95°C for 30 s and 60°C for 30 s for 45 cycles. The mRNA expression of FASN, ACC1, SCD1, HMGCR, DGAT, PGP and RPS6 a housekeeping gene used as an internal control, was determined. The gene expression was normalized to the housekeeping gene using the 2ΔΔCT method.

Lipidomic analysis. MDA-MB-231 cells were spread in a 6-well plate at a density of 5×105 cells/well and allowed to adhere overnight. After reaching the maximum confluence, cells were treated with 25 μM of 4Cone, 100 nM of TXT or their combination for 24 h at 37°C. Then, cells were lysed using M-PER. Protein concentrations were determined using a BCA assay (Sigma Aldrich), according to the manufacturer’s instructions. Then, extraction buffer (methanol/chloroform, 2/1, v/v, 800 μl) was added to sample homogenates, vortex-mixed and centrifuged for 10 min at 10,000 g for 10 min at 4°C. The supernatant was removed and dried under a gentle stream of nitrogen. Dried samples were then reconstituted with 100 μl of an isopropanol/acetonitrile/water mixture (60/35/5, v/v/v) and 10 μl were injected into the liquid chromatography-high-resolution mass spectrometry (LC-HRMS) system. Six replicates for each treated group were analyzed. Lipid separation and detection were achieved on an H-Class UPLC system coupled to a Synapt™ G2 HDMS Q-TOF mass spectrometer equipped with a Z-spray interface for electrospray ionization (Waters Corporation, Milford, MA, USA) as described previously (22, 23).

Processing of lipidomic data. Data acquisition and processing were achieved using MassLynx® and MakerLynx® software (version 4.1, Waters Corporation). Peak detection, integration, alignment, and normalization were performed using the following optimized parameters: mass window, 0.05 Da; retention time window, 0.1 min; peak width at 5% height, 12 s; peak-to-peak baseline noise, 500 (positive ionization) and 100 (negative ionization); smoothing, yes (3 points, Gaussian mode); marker intensity threshold, 500 counts (positive ionization) and 100 counts (negative ionization); noise elimination level, 6 (positive ionization) and 3 (negative ionization); deisotope data function, on; and replicate% minimum, 50%. During data processing, each specific signal, corresponding to a putative marker, was normalized to the total intensity of the full response measured in the sample. The mean response obtained over the six replicates was then calculated, constituting the raw data. Global lipid identification was realized with the LIPID MAPS database (LIPID Metabolites and Pathways Strategy). Lipid identification was first based on the exact mass measurement, considering the specific adducts formed during ionization. Fragmentation patterns obtained at high energy levels in both positive and negative modes were then used for structure elucidation. Several representative standards (at least four per lipid family) were used for confirmation.

Statistical analysis. Data were analyzed using nested one-way ANOVA, Two Way Repeated Measure ANOVA and student t-test using GraphPad Prism 8 (GraphPad Software, Boston, MA, USA). Brown-Forsythe correction was applied to test statistical difference between treated and control group. Principle component analysis was performed to evaluate distribution by using the R software. Tukey’s multiple comparison test method was applied to determine the mean of all possible pair of treatment. p-Values <0.05 were considered significant.

Results

4Cone alone or the combination 4Cone/TXT reduces cancer cell viability. Using the MTT assay, we first examined the effect of 4Cone alone, TXT alone or their combination on the viability of the MDA-MB-231 cells. MTT assays showed that 4Cone alone or in combination with TXT significantly lowered the viability of the MDA-MB-231 cell line compared to control. As illustrated in Figure 1, 4Cone alone showed a 50% and 70% viability reduction after 24 and 48 h of treatment, respectively. However, the combination 4Cone/TXT demonstrated a stronger reduction in cell viability (up to 75% in 24 h and up to 85% in 48 h).

Figure 1.
Figure 1.

Effect of 4-cholesten-3-one (4Cone) alone, docetaxel (TXT) alone or their combination (4Cone/TXT) for 24 and 48 h on MDA-MB-231 cell viability determined using the MTT assay. The data represent the mean±the standard deviation as compared with control. Statistical analysis was performed by two way repeated measure ANOVA followed by Tukey correction. ***p<0.001 compared to the control group.

4Cone alone or the combination 4Cone/TXT reduces cancer cell migration. Using the scratch wound healing test, we next assessed the impact of 4Cone on the migration of MDA-MB-231 cells during a 48 h treatment period. Compared with the treated group, cells without treatment gradually decreased the wound’s width (Figure 2A). After 48 h, the percentage of covered area was 40% relative to the untreated cells, indicating that 4Cone treatment decreased cell migration (Figure 2B). However, the effects of the chemotherapy medication TXT alone or in combination with 4Cone could not be ascertained because 90% of the cells detached during the observation period.

Figure 2.
Figure 2.

Effect of 4-cholesten-3-one (4Cone) on the migration of TNBC MDA-MB231 cells assayed using the wound healing assay. Cells were treated with 25 μM of 4Cone for 48 h. A) Images captured after 48 h of incubation. B) The quantitative analysis of the wound area using ImageJ software after 48 h. The width of scratch was calculated as a percentage of the initial gap. The data represent the mean±the standard deviation compared with the control. Statistical analysis was performed using unpaired t test. **p<0.01.

Down regulation of lipogenic genes by 4Cone alone or the combination 4Cone/TXT treatment. To examine whether the anti-migration and anti-proliferative effect were associated with lipid metabolism in MDA-MB-231 cells, we used real-time quantitative PCR to assess the mRNA expression of important lipogenic enzymes involved in the pathway of de novo synthesis of fatty acids, cholesterol, and triglycerides. As displayed in (Figure 3A) 4Cone alone, TXT alone or their combination significantly reduced the mRNA expression of fatty acid synthase (FASN) by 38, 50 and 20%, respectively, acetyl-CoA carboxylase (ACC) by 56, 57 and 30%, respectively, and stearoyl CoA desaturase 1 (SCD1) by 47, 47, and 25%, respectively. The expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), a key enzyme of the cholesterol synthesis pathway, was decreased significantly by 30% for 4Cone alone, by 40% for TXT alone and by 10% for the combination 4Cone/TXT (Figure 3B). Furthermore, 4Cone alone decreased the expression of diacylglycerol O-acyltransferase (DGAT) gene by 60%, which encodes for a key enzyme that converts diglyceride into triglyceride. However, TXT alone increased its expression up to 40% when compared to the control group. Interestingly, the combination treatment decreased the expression by 40% (Figure 3C). Moreover, Figure 3D illustrates that the expression of P-glycoprotein 1 (PGP), which is highly expressed in chemo resistance tumor cells, was reduced by 60% following treatment with 4Cone and 20% following treatment with the combination 4Cone/TXT.

Figure 3.
Figure 3.

Effect of 4-cholesten-3-one (4Cone) alone, docetaxel (TXT) alone or their combination (4Cone/TXT) on the expression of key lipogenic genes after 24 h of treatment. A) Expression levels of key fatty acid synthesis genes. B) Expression levels of cholesterol synthesis gene. C) Expression level of triglyceride synthesis gene. D) Genes involved in the chemo drug efflux mechanism. The data represent the mean±the standard deviation compared with the control. Statistically significant differences are indicated as ***p<0.001 and **p<0.01, analyzed through multiple comparisons (Tukey’s single-step test).

4Cone alone or the combination 4Cone/TXT changes the lipid profile. A total of 875 lipid species were identified using mass spectrometer. As illustrated in Figure 4, the relative abundance of ceramides (Cer) was higher in the cells treated with TXT alone or the combination 4Cone/TXT than in the control group. However, as compared to the control, there was no significant difference for the cells treated with 4Cone alone. In terms of sphingomyelins (SM), the opposite pattern was observed in the TXT group. The percentage of detected SM was lower in the TXT group compared to the control group. However, it increased in 4Cone group, whereas no significant difference was observed regarding the combination group. TG were found to be especially abundant in cells treated with TXT. However, 4Cone decreased the percentage of TG compared to the control group. The changes in monoglycerides (MG) and diglycerides (DG) were not different between the treated and control groups. In comparison to the control, the glycerophosphatidylcholine (PC) subtype was slightly lower in the single treatment group but did not change significantly in the combination group. The 4Cone and combination groups showed an increase in the percentage of cholesteryl esters (CE); nevertheless, the TXT-treated group showed no change in comparison to the control.

Figure 4.
Figure 4.

Intensity-based percentile of each lipid subclass in groups treated with 4Cone, TXT, 4Cone/TXT and control groups. Intensity percentage to the sum of all detected lipid species was calculated. Glycerophosphatidylcholine (PC), glycerophosphatidylethanolamine (PE), glycerophosphatidylglycerol (PG), glycerophosphatidylserine (PS), glycerophosphatidylinositol (PI), glycerophosphatidic acid (PA), lysoglycerophosphatidylcholine (LPC), lysoglycerophosphatidylethanolamine (LPE), cholesterol ester, monoglyceride (MG), sphingomyelins (SM), diglyceride (DG), triaglyceride (TG), ceramide (Cer).

Based on the changes in lipid abundance in control and treated cells, the levels of certain lipids within each lipid class were investigated (Figure 5). In the sphingolipid family, Cer [Cer (42:1) and Cer (42:2)] were significantly up-regulated in the TXT and combination groups compared to the control group. However, the 4Cone group did not show any significant variation in the observed species (Figure 5A). As expected, the opposite effect was observed for SM [SM (40:1), SM (42:1), and SM (42:2)] that were down regulated in the TXT and combination groups (4Cone/TXT) compared to the control group. In the acyl glycerol family, TG with saturated and monosaturated fatty acid [TG (52:0), TG (52:1), TG (52:2), TG (54:1), and TG (56:1)] were significantly up-regulated in cells treated with TXT compared to the control group (Figure 5B). Cells treated with 4Cone showed a significant reduction in TG levels. The combination treatment (4Cone/TXT) also reduced TG levels but the difference was not significant compared to the control group (Figure 5C). In the glycerophospholipid family, PC with polysaturated fatty acid [PC (38:5) and PC (40:6)] were significantly up-regulated in the TXT and combination group, whereas 4Cone had no significant effect. Furthermore, 4Cone up-regulated PC (42:0), whereas the other treatments (TXT alone and in combination with 4Cone) had no significant effect. None of the treatments showed significant up-regulation of PE (40:1) compared to the control (Figure 5D).

Figure 5.
Figure 5.

Normalized abundance of several lipid species. A) Two species of ceramide (Cer 42:1, Cer 42:2) and three species of sphingomyelin (SM 40:1, SM 42:1, SM 42:2). B) Selected species of triglyceride (TG 52:0, TG 52:1, TG 52:2, TG 54:1, 56:1) subclasses. C) Represents (PC 38:5, PC40:6, PC 42:0) species from glycerophosphatidylcholine subclass and (PE 40:1) from phosphatidylethanolamine subclass. The data represent the mean±the standard deviation compared with the control. Statistical analysis was performed by two way repeated measure ANOVA followed by Tukey correction. *p<0.05, **p<0.01, ***p<0.001.

Discussion

Since 4-cholesten-3-one (4Cone) exhibits anticancer properties by regulating cell growth, apoptosis, and metabolism (18, 19, 21), we questioned whether the alteration of the lipid profile was related to the anticancer effects. In this study, we investigated whether 4Cone could alter the lipid profile of MDA-MB-231, TNBC cells. Furthermore, examined the impact of 4Cone in combination with the anti-cancer chemo drug TXT in order to identify a better therapeutic combination.

In the current investigation, we discovered that 4Cone, either alone or in combination with TXT, have cytotoxic effects on MDA-MB-231 cells, resulting in a considerable reduction in cell viability following treatments for both 24 and 48 h. Similar results have been found in several studies. The MCF7 and MDA-MB-231 cell lines were subjected to treatment with different concentrations of various cytotoxic agents, including 25 μg/ml of 4Cone (21). Numerous studies have reported that TXT may disrupt microtubules to stop the cell cycle, which results in cell death and lower viability (24). The results of wound healing assay showed that 4Cone significantly reduces the cell migration in line with previous studies, which revealed that 4Cone has some anti-migration effects on MCF-7 and MDA-MB-231 cells (21).

Numerous reports indicate that the administration of pharmacological inhibitors of lipogenic enzymes involved in the lipid synthesis pathways substantially suppresses tumor development (25-27). Dysregulated metabolism of fatty acids, particularly aberrantly stimulated de novo fatty acid synthesis, has been linked to numerous malignant features of breast cancer (28, 29). Furthermore, the de novo cholesterol synthesis pathway, sometimes referred to as the mevalonate pathway, contains enzymes that are reported to be over-expressed in a number of breast cancer subtypes (30). The inhibition of a crucial enzyme in the de novo fatty acid and cholesterol pathways has been associated with the inhibition of invasion, migration, and proliferation of breast cancer cells (25-28, 30). According to our findings, the expression of FASN, ACC, SCD1, and HMGCR enzymes implicated in de novo fatty acid synthesis pathway as well as the cholesterol synthesis was down-regulated by 4Cone or its combination with TXT. Our results again showed that, while 4Cone and 4 Cone/TXT treated cancer cells showed a reduction in DGAT, TXT-treated cells showed greater expression of this gene, which is linked to the route for triglyceride (TG) formation, which is the primary source of energy storage in cancer cells (31). Based on our findings, 4Cone can be considered a potential therapy to suppress the expression of DGAT, which can control the production of triglycerides that rise in TXT-treated cells. Furthermore, over-expression of the xenobiotic efflux pump permeability glycoprotein (PGP) has been widely examined in TXT resistant cells, limiting the effectiveness of taxanes that block cell circle progression and induce apoptosis in cancer (32). In our study, we discovered that TXT in combination with 4Cone significantly reduces the expression of PGP compared to the control, implying that the combination may have the potential to improve the usefulness of taxanes in TXT resistant cells by limiting the expression of the xenobiotic efflux transporter PGP.

In order to determine whether the changes observed in the transcriptome also affect the lipidome, we examined major lipid classes that are crucial for cell membrane function, cell signaling, and energy storage. According to this study, 4Cone increases the percentage of SM while decreasing the percentage of Cer. However, TXT and the combination 4Cone/TXT increased Cer abundance while lowering that of SM. Few studies have demonstrated that increased production of specific Cer is associated with the development of various malignancies (33), but another study found that TXT elevates Cer levels, causing apoptosis (34). Furthermore, multiple studies have found that SM hydrolyzes to Cer, which inhibits cancer cell motility, migration, and metastasis (35). Due to the complexity, it is still unclear how Cer and SM contribute to the microenvironment to alter the phenotype of cancer cells (33-36). Furthermore, cancer cells treated with 4Cone or 4 Cone/TXT showed decreased TG levels, while those of TG increased in the TXT-treated cells. This outcome is consistent with a recent case study (37) that found patients with breast cancer had elevated TG levels after starting TXT. According to that study, TXT elevates TG levels, which may facilitate the growth of cancer cells by allowing them to absorb energy. Furthermore, increased TG may have a detrimental effect on the cardiovascular system and other metabolic processes (38). Based on our findings, 4Cone can lower TG levels in TXT-treated cells, which can contribute to the control of various metabolic disorders. Furthermore, we attempted to identify and quantify distinct subclasses of glycerophospholipids, which comprise the cellular membrane bilayer (39). The levels of detected PC reduced in all treated groups compared to the control. TXT had slightly higher levels of PE than the control group, but there was no significant difference between the 4Cone and combination groups. According to the findings of previous studies, PC and PE levels rise in TNBC cells compared to other types of cancer, which has also been considered a characteristic of TNBC (39, 40). Some drugs have been demonstrated to influence lipid metabolism, specifically phospholipid levels, to create anti-tumorigenic characteristics (41). Because of the complexity of the synthesis route and position in the cellular membrane, the role of the glycerophospholipid family in cancer cells remains unclear (39-41). Finally, the increase of Cholesteryl esters in 4cone and combination groups indicated a definite esterification brought on by the presence of sterol metabolites (4Cone) in the cell. According to our findings, TNBC cells (MDA-MB-231 cells) treated with 4Cone alone or its combination with TXT exhibited a variety of lipid metabolite alterations. Further research is necessary to fully understand their role in cancer progression or cell apoptosis because of the variety and complexity of the metabolic pathways controlling their biosynthesis and catabolism.

Conclusion

In this study, 4Cone alone and in combination with TXT have shown anti-proliferative and anti-migration activity in the TNBC cell line MDA-MB-231 through the reduction of the expression of key lipogenic genes. Additionally, we performed lipidomic analysis to examine the alteration of lipid profile in this cancer cell line by both treatments. Our results suggest that lipidome changes are probably linked to the effects of both drugs on lipid metabolism in cancer cells. One hypothesis for the increased effects of TXT in the presence of 4Cone on MDA-MB 231 cells is the reduced expression of the PGP transporter and DGAT, which could be attributed to 4Cone. Moreover, it can be hypothesized that 4Cone alone or the combination 4Cone/TXT have diverse effects on the MDA-MB-231 breast cancer cell. Taking everything into account, 4Cone, alone or in combination with TXT, affects TNBC’s lipid metabolism, which may have significance for the development of innovative cancer therapeutics. Further research on lipid species in breast cancer cells, and the enzymes that synthesize or degrade them in the presence of drugs, is needed to explain their involvement in carcinogenesis.

Acknowledgements

The Authors would like to thank la Ligue Contre le Cancer, France for funding the project and Erasmus Mundus ACES+ for financing Sristy Saha.

Footnotes

  • Authors’ Contributions

    Hassan Nazih: Conceived the concept designed the study and edited the manuscript. Sristy Saha: Carried out the experiments, analyzed the data and wrote the manuscript. Mikaël Croyal: Conducted, analyzed the lipidome studies and edited the manuscript. Jean-Michel Huvelin: Carried out the experiments.

  • Conflicts of Interest

    The Authors have no conflicts of interest to declare in relation to this study.

  • Funding

    This study was funded by la Ligue Contre le Cancer, France (Comité 44 de la Loire Atlantique et Comité 53 de Mayenne).

  • Received May 14, 2024.
  • Revision received June 6, 2024.
  • Accepted June 7, 2024.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

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4-Cholesten-3-one Modified the Lipidome of MDA-MB-231 Cells and Potentiated the Effect of Docetaxel
SRISTY SAHA, MIKAËL CROYAL, JEAN-MICHEL HUVELIN, HASSAN NAZIH
Anticancer Research Aug 2024, 44 (8) 3277-3285; DOI: 10.21873/anticanres.17146
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