A multi-omics approach reveals differences in toxicity and mechanisms in rice (Oryza sativa L.) exposed to anatase or rutile TiO2 nanoparticles.

Titanium dioxide nanoparticles (TiO2 NPs) have been widely used in agriculture, which increased the risk to soil-plant systems. Studies have demonstrated that TiO2 NPs can induce phytotoxicity. However, the toxicity mechanisms, particularly under the stress of TiO2 NPs with different crystalline forms, remain inadequately reported. In this study, we combined transcriptomics and metabolomics to analyze the toxicity mechanisms in rice (Oryza sativa L.) under the stress of anatase (AT) or rutile (RT) TiO2 NPs (50 mg/kg, 40 days). The length (decreased by 1.1-fold, p = 0.021) and malondialdehyde concentration (decreased by 1.4-fold, p = 0.0027) of rice shoots was significantly reduced after AT exposure, while no significant changes were observed following RT exposure. Antioxidant enzyme activities were significantly altered both in the AT and RT groups, indicating TiO2 NPs induced rice oxidative damage (with changes of 1.1 to 1.4-fold, p < 0.05). Additionally, compared to the control, AT exposure altered 3247 differentially expressed genes (DEGs) and 56 significantly differentially metabolites in rice (collectively involved in pyrimidine metabolism, TCA cycle, fatty acid metabolism, and amino acid metabolism). After RT exposure, 2814 DEGs and 55 significantly differentially metabolites were identified, which were collectively involved in fatty acid metabolism and amino acid metabolism. Our results indicated that AT exposure led to more pronounced changes in biological responses related to oxidative stress and had more negative effects on rice growth compared to RT exposure. These findings provide new insights into the phytotoxic mechanisms of TiO2 NPs with different crystalline forms. Based on the observed adverse effects, the study emphasizes that any form of TiO2 NPs should be used with caution in rice ecosystems. This study is the first to demonstrate that AT is more toxic than RT in paddy ecosystems, providing crucial insights into the differential impacts and toxic mechanisms of TiO2 NPs with different crystalline forms. These findings suggest prioritizing the use of RT when TiO2 NPs are necessary in agricultural development to minimize toxicity risks.

Presence of N, N'-Substituted p-Phenylenediamine-Derived Quinones in Human Urine.

Human exposure to various N,N'-substituted p-phenylenediamine-derived quinones (PPDQs) has been of increasing concern. Recent studies have examined N-phenyl-N'-(1,3-dimethylbutyl)-p-phenylenediamine-derived quinone (6PPDQ) in human urine to evaluate human exposure. However, other PPDQs in human urine have not been thoroughly investigated. This study analyzed six PPDQs in urine collected from 149 healthy individuals in Taizhou, China. All target PPDQs were detected, with 6PPDQ (mean 2.4 ng/mL,

Relationship Between Perfluoroalkyl Acids in Human Serum and Sjogren's Syndrome: A Case-Control Study of Populations in Hangzhou, China.

Exposure to perfluoroalkyl acids (PFAAs) has been found to elicit a range of detrimental effects on human health. However, limited research has investigated the impact of PFAAs on immunity and immune disorders such as Sjogren's syndrome, with existing studies yielding inconsistent results. This study was conducted in Hangzhou, China, with an initial cohort comprising 156 healthy individuals and 162 patients diagnosed with Sjogren's syndrome. We quantified serum levels of PFAAs and examined associations between PFAAs and both susceptibilities to the development of Sjogren's syndrome and immune marker concentrations. Nine PFAAs were frequently detected in the serum, with perfluorooctanoate (PFOA) exhibiting the highest concentration, followed by perfluorooctanesulfonate (PFOS). Exposure to PFOA and perfluorotridecanoate (PFTrDA) was inversely associated with the disease. Furthermore, a negative correlation between PFOA and C-reactive protein (CRP) was observed. These findings suggest that exposure to specific PFAAs may impact the immune system and potentially influence the development of Sjogren's syndrome.

2-Mercaptobenzothiazole-derived vulcanization accelerators in urine samples from Chinese adults.

Studies have discovered wide presence of 2-mercaptobenzothiazole (2-MBT) and 2-MBT-derived vulcanization accelerators (MVAs) in household dust samples, suggesting that these chemicals may have been pervasive in the environment. However, despite the potential for human exposure, the presence of MVAs in human urine, a common matrix used for assessing exposure to environmental chemicals, has not been thoroughly investigated. The current study comprehensively analyzed 11 kinds of MVAs in urine samples from the recruited general population (n = 197) living in Taizhou city, China. Five kinds of MVAs were detectable in >50 % of human urine samples. This indicates the widespread exposure to these vulcanization accelerators among the general population. The predominant target analytes in human urine were 2-MBT and 2,2'-dithiobisbenzothiazole (MBTS), with the mean urinary concentrations of 2.7 ng/mL (range

Nutrient competition between female cones and young seeds in spring affects the physiological dropping and nut-setting rate in Torreya grandis.

Physiological seed drop is a recognized phenomenon in economic forest, caused by the abscission of developing seeds due to intergroup competition for resources. However, little is known about the resource allocation dynamics in species exhibiting a biennial fruiting cycle, where interactions occur not only among seeds of the same year but also between reproductive structures from consecutive years. In this study, we explored the dynamics of resource allocation in Torreya grandis, a nut crop with a prototypical two-year seed development pattern. We implemented thinning treatments of 0%, 30%, and 60% on female cones and/or immature seeds during the spring, targeting various stages of development both pre- and post-pollination. Our findings reveal a pronounced resource competition in Torreya, evidenced by a natural seed-setting rate of merely 9.4%. Contrary to expectations, seed thinning did not lead to an obvious increase in nut-setting rates, whereas a substantial increase to 20.5% was observed when female cones were thinned by 60% at 20 days before pollination. The cone thinning treatment appears to have influenced seed development through positive cytokinin and negative abscisic acid effects. This indicates that intergroup competition between female cones and nuts is a more significant factor in seed drop than inner nut competition, and there seems to be an interaction between the two groups. We demonstrate that, in Torreya with biennial seed development, it is the competition between female cones and immature seeds that is important. This insight expands our comprehension of the physiological mechanisms governing seed drop in biennial fruiting species and managing the reproductive organ load to optimize nutrient allocation.

Identification of Primary Organophosphate Esters Contributing to Enhanced Risk of Gestational Diabetes Mellitus Based on a Case-Control Study.

Epidemiological studies on associations of organophosphate ester (OPE) exposure and gestational diabetes mellitus (GDM) risk, which remain rare and inconclusive, were carried out with a case-control population comprising 287 GDM and 313 non-GDM pregnant women recruited from Tianjin. The GDM group suffered distinctly higher serum concentrations of tri-n-butyl phosphate (TNBP), tri(2-butoxyethyl) phosphate (TBOEP), triphenyl phosphate (TPHP), tri-iso-propyl phosphate (TIPP), and tri(1-chloro-2-propyl) phosphate (TCIPP) than the healthy control group (p < 0.001). Traditional analysis methods employed for either individual or mixture effects found positive correlations (p < 0.05) between the concentrations of five OPEs (i.e., TNBP, TBOEP, TPHP, TIPP, and TCIPP) and the incidence of GDM, while 2-ethylhexyl diphenyl phosphate, tri(1-chloro-2-propyl) phosphate, and bis(2-ethylhexyl) phosphate exhibited opposite effects. Three machine learning methods considering the concurrence of OPE mixture exposure and population characteristics were applied to clarify their relative importance to GDM risk, among which random forest performed the best. Several OPEs, particularly TNBP and TBOEP ranking at the top, made greater contributions than some demographical characteristics, such as prepregnancy body mass index and family history of diabetes, to the occurrence of GDM. This was further validated by another independent case-control population obtained from Hangzhou.

BoMYBL2b, an R3-MYB transcription factor, inhibits anthocyanin accumulation via directly repressing BoDFR1 gene transcription in kale.

Kale is a globally popular edible and ornamental plant with colorful inner leaves. Anthocyanin is one of the most major pigments for kale. However, the underlying regulatory mechanism for the accumulation of anthocyanin in kale has not been well understood to date. Here, we characterized an R3-MYB transcription factor, BoMYBL2b, whose mRNA was missing in the anthocyanin-rich kale inbred line, but abundant in the anthocyanin-less line. Knockdown of BoMYBL2b enhanced the anthocyanin accumulation. Y1H and DLR assays showed that BoMYBL2b directly inhibited the BoDFR1 gene expression by binding to the MRE sites on the ProBoDFR1. Besides, we found that BoMYBL2b did not interact with the bHLH proteins, indicating that the BoMYBL2b might not be part of the MBW complex. BoMYBL2b expression was significantly repressed by BoMYB1R1 under high light and cold stimulus, suggesting the possibility that BoMYB1R1-BoMYBL2b module might act as a cross-node for integrating light and temperature signals to regulate anthocyanin accumulation in kale. These findings provided a theoretical basis for molecular breeding of new kale cultivars and a reference for understanding the regulatory mechanism of anthocyanin accumulation in Brassica.

Association between Serum 6:2 Chlorinated Polyfluorinated Ether Sulfonate Concentrations and Lung Cancer.

6:2 chlorinated polyfluorinated ether sulfonate (6:2 Cl-PFESA) exhibits pronounced estrogenic effects, potentially influencing the etiology of lung cancer. This study assessed the potential associations between serum concentrations of 6:2 Cl-PFESA and lung cancer risk at the population level. Odds ratios (ORs) for lung cancer across serum 6:2 Cl-PFESA quartiles were assessed using conditional logistic regression. Additionally, we investigated potential effect modification by various confounding factors. Elevated serum levels of 6:2 Cl-PFESA were consistently associated with an increased risk of lung cancer in both the crude model (OR = 1.62, 95% CI: 1.08-2.42, p = 0.018) and the adjusted model (OR = 1.59, 95% CI: 1.06-2.39, p = 0.026). Stratified analyses revealed that elevated serum levels of 6:2 Cl-PFESA were associated with increased risk estimates of lung cancer among males (adjusted OR = 2.04, 95% CI: 1.19-3.51, p = 0.006), smokers (adjusted OR = 2.48, 95% CI: 1.25-4.89, p = 0.003), and drinkers (adjusted OR = 2.20, 95% CI: 0.94-5.16, p = 0.049). The results of this study imply that exposure to 6:2 Cl-PFESA at levels considered environmentally relevant may be linked to an elevated risk of developing lung cancer.

Discovery of perfluoroalkyl sulfonyl quaternary ammonium substances in the environment and their environmental behaviors.

A variety of per- and polyfluoroalkyl substances (PFASs) have been released into the environment via wastewater treatment plant (WWTP) effluent, with current target and nontarget analytical methods typically focusing on negatively ionized PFASs while largely overlooking positively ionized ones. In this study, five cationic PFASs, perfluoroalkyl sulfonyl quaternary ammonium substances (PFAQASs), were first identified in surface water impacted by the WWTP effluent, applying a metabolomics-based nontarget analysis method. Environmental behaviors of identified novel PFAQASs were further investigated. In surface water, sediment, and fish (Coilia mystus) samples collected from the Yangtze River, 8:3 PFAQA was consistently the predominant PFAQASs, with the mean concentrations of 90 ng/L (< LOD-558 ng/L), 92 ng/g dw (< LOD-421 ng/g dw), and 2.3 ng/g ww (< LOD-4.6 ng/g ww), respectively. This study highlights the necessity to discover other cationic PFASs in the environment. Among PFAQASs, 8:4 PFAQA (4.2, range 3.4 - 4.6) had the highest mean sediment-water partitioning coefficient (log Koc), followed by 8:3 PFAQA (3.9, 2.8 - 4.5) and 6:3 PFAQA (3.7, 3.3 - 4.1). The log Koc of PFAQASs showed a general increase trend with the increasing carbon chain length. Mean bioaccumulation factor (BAF) values of PFAQASs calculated in the collected fish from the Yangtze River ranged from 1.9 ± 0.32 (4:3 PFAQA) to 2.9 ± 0.34 (8:4 PFAQA). The mean BAF values of PFAQASs generally increased with the carbon chain length. Further studies are warranted to elucidate the environmental fate, potential toxicity, and human exposure implications for these identified novel PFASs.

Bisphenol S and Its Chlorinated Derivatives in Indoor Dust and Human Exposure.

Bisphenol S (BPS), an environmental endocrine disruptor, has been identified in global environmental matrices. Nevertheless, limited studies have investigated the presence of chlorinated analogues of BPS (Clx-BPSs) with potential estrogenic activities in environmental matrices. In this study, the occurrence of BPS and five types of Clx-BPSs was characterized in indoor dust (n = 178) from Hangzhou City. BPS was measurable in 94% of indoor dust samples, with an average level of 0.63 µg/g (

Occurrence of bisphenol analogues and their conjugated metabolites in foodstuff.

Bisphenol analogues (BPs) are prevalent in diverse foodstuff samples worldwide. However, the occurrence of conjugated bisphenol A (BPA) and bisphenol S (BPS) metabolites in foodstuff remains poorly understood. This study analyzed eight BPs, and four conjugated BPA and BPS metabolites, in three animal-derived foodstuff and five plant-derived foodstuff samples from China. Results showed that fish foodstuff (9.7 ng/g ww) contained the highest mean concentration of BPA, followed by rice (5.1 ng/g ww) and beans foodstuff (3.6 ng/g ww). BPA-sulfate had higher mean concentrations than BPA-glucuronide in different foodstuff categories, except that in eggs foodstuff (p < 0.05). Compared with other foodstuff items, fish (3.4 ng/g ww) and vegetable (1.6 ng/g ww) foodstuff samples exhibited comparatively higher mean concentrations of BPS. Mean concentrations of BPS-sulfate were consistently higher than BPS-glucuronide in vegetables, meats, and fish foodstuff (p < 0.05). BPA contributed the major total dietary intake (DI) of BPs, with the mean DI of 435 ng/kg bw/day for women and 374 ng/kg bw/day for men, respectively. To our knowledge, this study is the first to investigate the occurrence of conjugated BPA and BPS metabolites in foodstuff, which enhances our comprehension of the origins of these conjugated metabolites in the human body.

Presence of benzotriazole ultraviolet stabilizers in human urine.

Health exposure to benzotriazole ultraviolet stabilizers (BUVSs) may pose diverse toxic impacts on health. Presently, the occurrence of BUVSs in human urine remains inadequately understood. This study analyzed 13 kinds of BUVSs in human urine (n = 182) from the general Chinese adult participants. Totally, nine BUVSs were measurable in these human urine samples. Among the detected BUVSs, 2-(2H-benzotriazol-2-yl)-p-cresol (UV-P) was the most predominant BUVS in the human urine, with the mean concentration of 1.6 µg/g creatinine (

6:2 Cl-PFESA, a proposed safe alternative for PFOS, diminishes the gemcitabine effectiveness in the treatment of pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC)/pancreatic cancer, is a highly aggressive malignancy with poor prognosis. Gemcitabine-based chemotherapy remains the cornerstone of PDAC treatment. Nonetheless, the development of resistance to gemcitabine among patients is a major factor contributing to unfavorable prognostic outcomes. The resistance exhibited by tumors is modulated by a constellation of factors such as genetic mutations, tumor microenvironment transforms, environmental contaminants exposure. Currently, comprehension of the relationship between environmental pollutants and tumor drug resistance remains inadequate. Our study found that PFOS/6:2 Cl-PFESA exposure increases resistance to gemcitabine in PDAC. Subsequent in vivo trials confirmed that exposure to PFOS/6:2 Cl-PFESA reduces gemcitabine's efficacy in suppressing PDAC, with the inhibition rate decreasing from 79.5 % to 56.7 %/38.7 %, respectively. Integrative multi-omics sequencing and molecular biology analyses have identified the upregulation of ribonucleotide reductase catalytic subunit M1 (RRM1) as a critical factor in gemcitabine resistance. Subsequent research has demonstrated that exposure to PFOS and 6:2 Cl-PFESA results in the upregulation of the RRM1 pathway, consequently enhancing chemotherapy resistance. Remarkably, the influence exerted by 6:2 Cl-PFESA exceeds that of PFOS. Despite 6:2 Cl-PFESA being regarded as a safer substitute for PFOS, its pronounced effect on chemotherapeutic resistance in PDAC necessitates a thorough evaluation of its potential risks related to gastrointestinal toxicity.

Conjugated metabolites of bisphenol A and bisphenol S in indoor dust, outdoor dust, and human urine.

Bisphenol A (BPA) and bisphenol S (BPS) have been widely spread in the global environment. However, for conjugated BPA and BPS metabolites, limited studies have investigated their occurrence in environmental matrices. We collected paired indoor and outdoor dust (n = 97), as well as human urine (n = 153) samples, from residential houses in Quzhou, China, and measured these samples for 8 conjugated BPA and BPS metabolites. Three BPA metabolites were found in collected indoor and outdoor dust, with BPA sulfate (mean 0.75 and 1.3 ng/g, respectively) and BPA glucuronide (0.13 and 0.26 ng/g) being more abundant. BPA conjugates accounted for a mean of 42 and 56% of total BPA (sum of conjugated BPA and BPA metabolites) in indoor and outdoor dust, respectively. BPS sulfate (mean 0.29 and 0.82 ng/g, respectively) had consistently higher concentrations than BPS glucuronide (0.13 and 0.27 ng/g) in indoor and outdoor samples. BPS conjugates contributed a mean 32% and 45% of total BPS (sum of BPS and BPS metabolites) in indoor and outdoor dust, respectively. Moreover, conjugated BPA and BPS metabolites in indoor or outdoor dust were not significantly correlated with those in urine from residents. Overall, this study first demonstrates the wide presence of conjugated BPA and BPS metabolites, besides BPA and BPS, in indoor and outdoor dust. These data are important for elucidating the sources of conjugated BPA and BPS metabolites in the human body.

A novel three-layer module BoMYB1R1-BoMYB4b/BoMIEL1-BoDFR1 regulates anthocyanin accumulation in kale.

Anthocyanin is an important pigment responsible for plant coloration and beneficial to human health. Kale (Brassica oleracea var. acephala), a primary cool-season flowers and vegetables, is an ideal material to study anthocyanin biosynthesis and regulation mechanisms due to its anthocyanin-rich leaves. However, the underlying molecular mechanism of anthocyanin accumulation in kale remains poorly understood. Previously, we demonstrated that BoDFR1 is a key gene controlling anthocyanin biosynthesis in kale. Here, we discovered a 369-bp InDel variation in the BoDFR1 promoter between the two kale inbred lines with different pink coloration, which resulted in reduced transcriptional activity of the BoDFR1 gene in the light-pink line. With the 369-bp insertion as a bait, an R2R3-MYB repressor BoMYB4b was identified using the yeast one-hybrid screening. Knockdown of the BoMYB4b gene led to increased BoDFR1 expression and anthocyanin accumulation. An E3 ubiquitin ligase, BoMIEL1, was found to mediate the degradation of BoMYB4b, thereby promoting anthocyanin biosynthesis. Furthermore, the expression level of BoMYB4b was significantly reduced by light signals, which was attributed to the direct repression of the light-signaling factor BoMYB1R1 on the BoMYB4b promoter. Our study revealed that a novel regulatory module comprising BoMYB1R1, BoMIEL1, BoMYB4b, and BoDFR1 finely regulates anthocyanin accumulation in kale. The findings aim to establish a scientific foundation for genetic improvement of leaf color traits in kale, meanwhile, providing a reference for plant coloration studies.

Discovery of novel benzotriazole ultraviolet stabilizers in surface water.

The comprehensive understanding of the occurrence of benzotriazole UV stabilizers (BZT-UVs) in environmental surface water is imperative due to their widespread application and potential aquatic toxicity. We conducted an analysis of 13 traditional BZT-UVs in surface water samples collected from Taihu Lake (TL, n = 23) and Qiantang River (QR, n = 22) in China. The results revealed that 5­chloro-2-(3,5-di-tertbutyl-2-hydroxyphenyl)-benzotriazole (UV-327) was consistently the predominant BZT-UV in water samples from TL (mean 16 ng/L; detection frequency 96 %) and QR (14 ng/L; 91 %). Furthermore, we developed a characteristic fragment ion-based strategy to screen and identify unknown BZT-UVs in collected surface water, utilizing a high-resolution mass spectrometer. A total of seven novel BZT-UVs were discovered in water samples, and their chemical structures were proposed. Four of these novel BZT-UVs were further confirmed with standards provided by industrial manufacturers. Semi-quantitative analysis revealed that among discovered novel BZT-UVs, 2-(2­hydroxy-3­tert­butyl­5-methylphenyl)-benzotriazole was consistently the predominant novel BZT-UV in TL (mean 4.1 ng/L, detection frequency 70 %) and QR (2.8 ng/L, 77 %) water. In TL water, the second predominant novel BZT-UV was 2-(3-allyl-2­hydroxy-5-methylphenyl)-2H-benzotriazole (mean 3.9 ng/L,

Conjugated bisphenol S metabolites in human serum and whole blood.

Studies have shown that bisphenol S (BPS) is mainly present as its conjugated metabolites in human blood. However, the distribution of conjugated BPS metabolites in different human blood matrices has not been characterized. In this study, paired human serum and whole blood samples (n = 79) were collected from Chinese participants, and were measured for the occurrence of BPS and 4 BPS metabolites. BPS was detectable in 49% of human serum (

Presence of carbazole and polyhalogenated carbazoles in human urine.

Exposure to carbazole (CZ) and polyhalogenated carbazoles (PHCZs) may pose a threat to human health, owing to their potential dioxin-like toxicity. Until now, the presence of these chemicals in the human urine from the general population is still unclear. Human urine samples (n = 210) were taken from the general population in Quzhou, China in this study, and were analyzed for CZ and 14 PHCZs. CZ and nine PHCZs were detected in collected human urine. CZ (detection frequency 100 %), 3-chlorocarbazole (3-CCZ; 88 %), 3,6-dichlorocarbzole (36-CCZ; 84 %), and 3-bromocarbazole (3-BCZ; 80 %) were more frequently detected. Among detected PHCZs, 3-CCZ (mean 0.49 ng/mL, < LOD-4.3 ng/mL) had comparatively higher urinary levels, followed by 3-BCZ (0.30 ng/L, < LOD-1.9 ng/mL) and 36-CCZ (0.20 ng/L, < LOD-1.4 ng/mL). Urinary concentrations of CZ in male participants (1.3 ± 0.26 ng/mL) were significantly (p < 0.05) higher than that in female participants (0.92 ± 0.24 ng/mL). No obvious trend in urinary concentrations with the age of participants was found for CZ and detected PHCZs. The mean daily excretion was found highest for CZ (31 ng/kg bw/day), followed by 3-CCZ (19 ng/kg bw/day) and 3-BCZ (8.5 ng/kg bw/day). This study provides the first data, to our knowledge, on the presence and levels of CZ and PHCZs in human urine, which is necessary for conducting the human exposure risk assessment.

Sediment-seawater partitioning, bioaccumulation, and biomagnification of perfluorobutane sulfonamide in marine environment.

Environmental occurrence of perfluorobutane sulfonamide (PFBSA) has only been recently discovered. The current knowledge regarding the occurrence and environmental behaviors of PFBSA in the marine environment is still relatively limited. In this study, PFBSA and other 37 poly- and perfluoroalkyl substances were analyzed in seawater (n = 43), sediment (n = 43), and marine fish (n = 176) samples collected from East China Sea and Antarctic Ocean. PFBSA was detected in > 90% of seawater from East China Sea and Antarctic Ocean, with the concentrations of 1.0 - 19 ng/L and < LOD-228 pg/L, respectively. The field-based mean log-transformed sediment-seawater partitioning coefficients of PFBSA were 1.6 ± 0.19 L/kg dw and 1.1 ± 0.19 L/kg dw in East China Sea and Antarctic Ocean, respectively, which are lower than that of perfluorooctanoate and perfluorooctane sulfonate. This indicates its long-range transport potential in global oceans with ocean currents. The mean log-transformed bioaccumulation factor values of PFBSA determined in the multiple species of whole-body marine fishes from East China Sea and Antarctic Ocean were 2.3 L/kg ww and 2.4 L/kg ww, respectively, which are comparable to that of perfluoroheptanoate (2.3 L/kg ww) in marine fishes from East China Sea. We did not observe an obvious biomagnification or biodilution of PFBSA along the marine food chain in East China Sea or Antarctic Ocean. This study provides the first data on the environmental behaviors of PFBSA in the marine environment.

Evidence of promoting effects of 6:2 Cl-PFESA on hepatocellular carcinoma proliferation in humans: An ideal alternative for PFOS in terms of environmental health?

Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are synthetic chemicals, encompassing compounds like perfluorooctane sulfonate (PFOS), which have widespread applications across various industries, including food packaging and firefighting. In recent years, China has increasingly employed 6:2 Cl-PFESA as an alternative to PFOS. Although the association between PFAS exposure and hepatocellular carcinoma (HCC) has been demonstrated, the underlying mechanisms that promote HCC proliferation are uncleared. Therefore, we aimed to investigate the effects and differences of PFOS and 6:2 Cl-PFESA on HCC proliferation through in vivo and in vitro tumor models. Our results reveal that both PFOS and 6:2 Cl-PFESA significantly contribute to HCC proliferation in vitro and in vivo. Exposure led to reduced population doubling times, enlarged cell colony sizes, enhanced DNA synthesis efficiency, and a higher proportion of cells undergoing mitosis. Furthermore, both PFOS and 6:2 Cl-PFES) have been shown to activate the PI3K/AKT/mTOR signaling pathway and inhibit necroptosis. This action consequently enhances the proliferation of HCC cells. Our phenotypic assay findings suggest that the tumorigenic potential of 6:2 Cl-PFESA surpasses that of PFOS; in a subcutaneous tumor model using nude mice, the mean tumor weight for the 6:2 Cl-PFESA-treated cohort was 2.33 times that observed in the PFOS cohort (p < 0.01). Despite 6:2 Cl-PFESA being considered a safer substitute for PFOS, the pronounced effects of this chemical on HCC cell growth warrant a thorough assessment of hepatotoxicity risks linked to its usage.