HPV-YAP1 oncogenic alliance drives malignant transformation of fallopian tube epithelial cells.

High grade serous ovarian carcinoma (HGSOC) is the most common and aggressive ovarian malignancy. Accumulating evidence indicates that HGSOC may originate from human fallopian tube epithelial cells (FTECs), although the exact pathogen(s) and/or molecular mechanism underlying the malignant transformation of FTECs is unclear. Here we show that human papillomavirus (HPV), which could reach FTECs via retrograde menstruation or sperm-carrying, interacts with the yes-associated protein 1 (YAP1) to drive the malignant transformation of FTECs. HPV prevents FTECs from natural replicative and YAP1-induced senescence, thereby promoting YAP1-induced malignant transformation of FTECs. HPV also stimulates proliferation and drives metastasis of YAP1-transformed FTECs. YAP1, in turn, stimulates the expression of the putative HPV receptors and suppresses the innate immune system to facilitate HPV acquisition. These findings provide critical clues for developing new strategies to prevent and treat HGSOC.

Transgenerational Effects and Mechanisms of Tributyltin Exposure on Neurodevelopment in the Male Offspring of Rats.

This study aimed to investigate the transgenerational effects of tributyltin exposure on rat neurodevelopment in male offspring and the potential mechanisms. Neonatal female rats were exposed to the environmental level of tributyltin and then mated with nonexposed males after sexual maturity to produce the F1 generation. The F1 generation (with primordial germ cell exposure) was mated with nonexposed males to produce nonexposed offspring (the F2 and F3 generations). Neurodevelopmental indicators and behavior were observed for the F1, F2, and F3 generations during postnatal days 1-25 and 35-56, respectively. We found premature eye-opening and delayed visual positioning in newborn F1 rats and anxiety and cognitive deficits in prepubertal F1 male rats. These neurodevelopmental impacts were also observed in F2 and F3 males. Additionally, F1-F3 males exhibited increased serotonin and dopamine levels and a loose arrangement of neurons in the hippocampus. We also observed a reduction in the expression of genes involved in intercellular adhesion and increased DNA methylation of the Dsc3 promoter in F1-F3 males. We concluded that tributyltin exposure led to transgenerational effects on neurodevelopment via epigenetic reprogramming in male offspring. These findings provide insights into the risks of neurodevelopmental disorders in offspring from parents exposed to tributyltin.

Estradiol protects female mice from hyperuricemia induced by PCB138 exposure.

Polychlorinated biphenyls (PCBs) are a type of persistent organic pollutant (POP). Our previous study demonstrated that exposure to 0.5-50 µg/kg bw PCB138 during postnatal days (PND) 3-21 led to elevated serum uric acid (UA) levels and kidney injury in adult male mice. Given that the prevalence of hyperuricemia (HUA) is significantly lower in women than in men, it is worth investigating whether POP-induced HUA and its secondary kidney injury have sexual dimorphism. Herein, we exposed female mice to 0.5-50 µg/kg bw PCB138 during PND 3-21, resulting in elevated serum UA levels, but without causing significant kidney damage. Concurrently, we found a negative correlation between serum 17ß-estradiol (E2) and serum UA levels. We also observed down-regulation of estrogen receptor (ER) protein levels in the kidneys of the PCB138-exposed groups. Furthermore, our study showed that E2 rescued the increased UA level and cytotoxicity caused by HUA in human renal tubular epithelial (HK-2) cells. Collectively, our findings suggest that E2 likely plays a crucial protective role in PCB138-induced HUA and kidney injury in female mice. Our research highlights the existence of sexual dimorphism in kidney injury secondary to HUA induced by POPs, which could provide guidance for individuals of different genders in preventing kidney injury caused by environmental factors.

Adolescent exposure to environmental level of PCBs (Aroclor 1254) induces non-alcoholic fatty liver disease in male mice.

Polychlorinated biphenyls (PCBs) are persistent organic pollutants found in various environmental media, and there is growing evidence that PCBs may contribute to the pathogenesis of non-alcoholic fatty liver disease (NAFLD). The purposes of this study were to investigate whether environmental level of Aroclor 1254 (a commercial mixture of PCBs) exposure to adolescent male mice could induce the development of NAFLD and the mechanisms involved. Twenty-one-day-old male C57BL/6 mice were exposed to Aroclor 1254 (0.5-500 µg/kg body weight) by oral gavage once every third day for 60 days. The results showed that exposure to Aroclor 1254 increased body weight and decreased the liver-somatic index in a dose-dependent manner. Aroclor 1254 administration increased lipid accumulation in the liver and induced the mRNA expression of genes associated with lipogenesis, including acetyl-CoA carboxylase 1 (Acc1), acetyl-CoA carboxylase 2 (Acc2) and fatty acid synthase (Fasn). Moreover, Aroclor 1254 decreased peroxisome proliferator-activated receptor alpha (PPARα) signaling and lipid oxidation. In addition, we found that Aroclor 1254 administration induced oxidative stress in mouse liver and elevated the protein level of cyclooxygenase 2 (COX-2), an inflammatory molecule, possibly via the endoplasmic reticulum (ER) stress inositol-requiring enzyme 1α-X-box-binding protein-1 (IRE1α-XBP1) pathway, but not the nuclear factor-κB (NF-κB) pathway. In summary, adolescent exposure to environmental level of PCBs stimulated oxidative stress, ER stress and the inflammatory response and caused NAFLD in male mice. This work provides new insight into the idea that adolescent exposure to environmental level of PCBs might induce the development of NAFLD under the regulation of ER stress in males.

Combined effects of ocean acidification and crude oil pollution on tissue damage and lipid metabolism in embryo-larval development of marine medaka (Oryzias melastigma).

Ocean acidification (OA) and crude oil pollution have been highlighted as some of the most pervasive anthropogenic influences on the ocean.In marine teleosts, early life-history stages are particularly vulnerable to disturbance by CO2-driven acidification as they lack pH-mediated intracellular regulation. Embryos exposed to trace levels of crude oil constituents dissolved in water exhibit a common syndrome of developmental abnormalities. So far, little is known about the combined effects of OA and crude oil on the early life history of marine fish. Eggs and larvae of the marine medaka (Oryzias melastigma) were treated with CO2 (1080 µatm atmospheric CO2), the water-soluble fraction (WSF) of crude oil (500 µg/L) and a CO2 (1080 µatm atmospheric CO2)/WSF (500 µg/L) mixture within 4 h after oviposition. Isolated and combined OA/WSF had no detectable effect on embryonic duration, egg survival rate and size at hatching. Histopathological anomalies of tissue and lipid metabolic disorder were significant when CO2 or WSF was given alone at 30 days of age. Combination of CO2 and WSF enhanced their toxicity compared to their separate administration. Since the early life-history stage of marine fish is thought to be impacted more heavily by increasing CO2 partial pressure (pCO2) levels and crude oil pollution, OA and crude oil pollution have the potential to act as an additional source of natural mortality.

Early life PCB138 exposure induces kidney injury secondary to hyperuricemia in male mice.

Polychlorinated biphenyls (PCBs) are a class of persistent organic pollutants (POPs) that have adverse effects on human health. However, the long-term health effects and potential mechanism of neonatal exposure to PCBs are still unclear. In this study, nursing male mice exposed to PCB138 at 0.5, 5, and 50 µg/kg body weight (bw) from postnatal day (PND) 3 to PND 21 exhibited increased serum uric acid levels and liver uric acid synthase activity at 210 days of age. We also found an increased kidney somatic index in the 50 µg/kg group and kidney fibrosis in the 5 and 50 µg/kg groups. Mechanistically, PCB138 induced mitochondrial dysfunction and endoplasmic reticulum (ER) stress, which might have led to inflammatory responses, such as activation of the NF-κB (nuclear factor kappa-B) and NLRP3 (NOD-like receptor protein 3) pathways. The inflammatory response might regulate renal fibrosis and hypertrophy. In summary, this study reports a long-term effect of neonatal PCB exposure on uric acid metabolism and secondary nephrotoxicity and clarifies the underlying mechanism. Our work also indicates that early life pollutant exposure may be an important cause of diseases later in life.

Screening of potential oestrogen receptor α agonists in pesticides via in silico, in vitro and in vivo methods.

In modern agricultural management, the use of pesticides is indispensable. Due to their massive use worldwide, pesticides represent a latent risk to both humans and the environment. In the present study, 1056 frequently used pesticides were screened for oestrogen receptor (ER) agonistic activity by using in silico methods. We found that 72 and 47 pesticides potentially have ER agonistic activity by the machine learning methods random forest (RF) and deep neural network (DNN), respectively. Among endocrine-disrupting chemicals (EDCs), 14 have been reported as EDCs or ER agonists by previous studies. We selected 3 reported and 7 previously unreported pesticides from 76 potential ER agonists to further assess ERα agonistic activity. All 10 selected pesticides exhibited ERα agonistic activity in human cells or zebrafish. In the dual-luciferase reporter gene assays, six pesticides exhibited ERα agonistic activity. Additionally, nine pesticides could induce mRNA expression of the pS2 and NRF1 genes in MCF-7 cells, and seven pesticides could induce mRNA expression of the vtg1 and vtg2 genes in zebrafish. Importantly, the remaining 48 out of 76 potential ER agonists, none of which have previously been reported to have endocrine-disrupting effects or oestrogenic activity, should be of great concern. Our screening results can inform environmental protection goals and play an important role in environmental protection and early warnings to human health.

Neonatal exposure to environment-relevant levels of tributyltin leads to uterine dysplasia in rats.

Endocrine-disrupting chemicals (EDCs) are natural/synthetic compounds that mimic or inhibit the biological actions of endogenous hormones. Studies have revealed that environmental estrogen, such as bisphenol A (BPA), causes developmental defects in the uterus. Tributyltin (TBT) is a typical environmental androgen. In this study, we aimed to explore the effect and mechanism of TBT on uterine development. Neonatal female rats were exposed to TBT (10 and 100 ng/kg bw) from postnatal days 1 to 16. BPA (50 µg/kg bw) was used as a positive control. Neonatal exposure to environmental concentrations of TBT resulted in pathological changes in the uterus, including thickening of the uterine luminal epithelium, a low density of glands, endometrial inflammation and fibrosis. Further, TBT affected the Wnt signaling pathway, which might mediate developmental disorders of the endometrial epithelial cells and glands in the uterus. TBT exposure also activated the NF-κB signaling pathway, which triggered inflammation. Moreover, TBT exposure upregulated the TGF-ß/Smads signaling pathway, possibly leading to endometrial fibrosis. In summary, our results demonstrate that neonatal exposure to an environment-relevant level of TBT leads to uterine dysplasia and provide potential molecular mechanisms. Our study is helpful for clarifying the effects of environmental androgens on the female reproduction system.

Generation and application of a Tg(cyp1a:egfp) transgenic marine medaka (Oryzias melastigma) line as an in vivo assay to sensitively detect dioxin-like compounds in the environment.

Large-range environmental pollution by dioxin and dioxin-like compounds (DLCs) is becoming a serious problem. To establish an in vivo method for the detection of DLCs in seawater, a Tg(cyp1a-12DRE:egfp) transgenic marine medaka (Oryzias melastigma) line was first developed with the modified cyp1a-12DRE promoter driving enhanced green fluorescent protein (EGFP) expression using Tol2 transgenesis technology. With increasing concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), the EGFP fluorescence intensity increased significantly. The Tg(cyp1a-12DRE:egfp) medaka possessed high sensitivity (limit of detection of 1 ng/L TCDD) and specificity and low background. This transgenic line is capable of detecting DLCs in environmental seawater in which the concentration of DLCs is at least 0.12207 ng/L TCDD after sample enrichment. The fluorescence-toxic equivalency (TEQ) values from EGFP intensity were closely correlated with the chemical-TEQ values obtained from chemical analyses. Furthermore, the Tg(cyp1a-12DRE:egfp) medaka can directly detect DLCs in seawater samples after a serious pollution accident and screen unknown aryl hydrocarbon receptor (AhR) agonists for risk assessment. For the first time, a convenient method has been established that sensitively and specifically responds to DLCs using the Tg(cyp1a-12DRE:egfp) marine medaka, which could be a highly efficient tool for detecting seawater DLCs in the future.

The protective effects of Nile tilapia (Oreochromis niloticus) scale collagen hydrolysate against oxidative stress induced by tributyltin in HepG2 cells.

Oxidative stress is regarded as one of the most important factors associated with many diseases, such as atherosclerosis, cancer, and diabetes. Various chemicals are released into the environment, causing environmental pollution. Importantly, many of them may cause damage to organisms through oxidative stress. In this work, we investigated the possible protective effects of Nile tilapia (Oreochromis niloticus) scale collagen hydrolysate (TSCH) (molecular weight approximately 4 kDa) against tributyltin (TBT)-induced oxidative stress in vitro. The results showed that pretreatment with TSCH protected against decreases in cell viability and changes in cell morphology in HepG2 cells exposed to TBT. Treatment with TSCH reduced the TBT-induced elevation in malondialdehyde (MDA) levels in HepG2 cells in a dose-dependent manner. Pretreatment with TSCH increased glutathione reductase (GR) and superoxide dismutase (SOD) activity. Moreover, TSCH decreased the expression of the proapoptotic protein Bax, reducing apoptosis. These results suggest that the protective mechanism of TSCH may be associated with its ability to scavenge MDA, increase antioxidant enzyme activity and downregulate the expression of Bax.

Generation of a Tg(cyp1a-12DRE:EGFP) transgenic zebrafish line as a rapid in vivo model for detecting dioxin-like compounds.

Dioxin-like compounds (DLCs) are extremely stable toxic organic compounds and can cause serious health risks. To develop a convenient biomonitoring tool for the detection of DLCs in the environment, we generated a transgenic line-Tg(cyp1a-12DRE:EGFP)-with a zebrafish cyp1a promoter recombined with multiple dioxin-responsive elements (DREs) that drive EGFP expression. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-induced EGFP expression was observed in the head cartilage (most sensitive), gut, otic vesicle, pectoral fin bud and eye of larvae. The lowest observed effect concentration of TCDD was estimated to be approximately 1 ng/L. Compared with existing zebrafish lines, our transgenic fish displayed comparable or even higher detection sensitivity to DLCs and could serve as an improved and rapid assay in an in vivo context. The Tg(cyp1a-12DRE:EGFP) transgenic zebrafish line also had higher stability for inducing EGFP expression (nearly 100% of our zebrafish induced EGFP at approximately 1 ng/L TCDD) than other lines. In addition, Tg(cyp1a-12DRE:EGFP) zebrafish could serve as a convenient and straightforward tool to assess potential cranial malformations and related health effects.

Protective effects of fucoxanthin and fucoxanthinol against tributyltin-induced oxidative stress in HepG2 cells.

Tributyltin (TBT) is a biocide extremely toxic to a wide range of organisms, which has been used for decades for industrial purposes. Fucoxanthin is a natural carotenoid that is isolated from seaweed, and fucoxanthinol is a major primary metabolite of fucoxanthin. Although fucoxanthin and fucoxanthinol have been reported to possess anti-oxidant activities in vitro, little is known as to whether they protect against TBT-induced oxidative stress in cultured cells. In the present study, the protective effect of fucoxanthin and fucoxanthinol against oxidative stress induced by TBT was investigated. The data showed that incubation of HepG2 cells with 0.2 µM TBT significantly increased cell apoptosis, whereas treatment with fucoxanthin or fucoxanthinol (3 µM) significantly recovered cell viability. In addition, fucoxanthinol treatment significantly decreased the intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) in HepG2 cells incubated with TBT. Moreover, fucoxanthin and fucoxanthinol markedly increased the expression level of Bcl-2/Bax. These results demonstrated that both fucoxanthin and fucoxanthinol effectively prevented cytotoxicity in HepG2 cells treated with TBT, and the protective effect was likely associated with decreased intracellular ROS and MDA and increased Bcl-2/Bax levels.