Cadmium (Cd), a heavy metal, is implicated in the development of estrogen receptor α (ERα)-positive breast cancers (BCs). However, controversy surrounds whether Cd is estrogenic or anti-estrogenic for the malignancy of in vivo animal models and human observational/epidemiological studies, a debate also presents in in vitro experiments. The development of ERα-positive BCs is stimulated by circulating 17β-estradiol (E2). Thus, potential biological interactions between E2 and Cd in the progression of ERα-positive BCs exist. Although the interactions between Cd and E2 at physiologically relevant levels (1 nM each) may not have been confirmed in vitro, it is likely to occur. Therefore, this study aims to investigate the interactions of chemicals in human BC MCF-7 cells (ERα-positive) using a sequential exposure system in which chemicals are added to cells every 24-48 h. Pretreatment with Cd, but not secondary treatment, interfered with E2-mediated oncogenic actions by inducing cell cycle arrest at the S phase. This was accompanied by changes in the expression of genes regulating the cell cycle checkpoint and upregulation of the tumor suppressor metallothionein 1F gene, which E2 suppressed. Paradoxically, ERα-mediated estrogenic pathways were upregulated. In conclusion, this study is the first to show that physiologically relevant levels of Cd may dampen E2-induced oncogenic events independent of the E2/ERα-mediated pathway.

To identify effective treatment modalities for breast cancer with acquired resistance, we first compared the responsiveness of estrogen receptor-positive breast cancer MCF-7 cells and long-term estrogen-deprived (LTED) cells (a cell model of endocrine therapy-resistant breast cancer) derived from MCF-7 cells to G-1 and 2-methoxyestradiol (2-MeO-E2), which are microtubule-destabilizing agents and agonists of the G protein-coupled estrogen receptor 1 (GPER1). The expression of GPER1 in LTED cells was low (~0.44-fold), and LTED cells displayed approximately 1.5-fold faster proliferation than MCF-7 cells. Although G-1 induced comparable antiproliferative effects on both MCF-7 and LTED cells (IC50 values of >10 µM), 2-MeO-E2 exerted antiproliferative effects selective for LTED cells with an IC50 value of 0.93 μM (vs. 6.79 μM for MCF-7 cells) and induced G2/M cell cycle arrest. Moreover, we detected higher amounts of β-tubulin proteins in LTED cells than in MCF-7 cells. Among the β-tubulin (TUBB) isotype genes, the highest expression of TUBB2B (~3.2-fold) was detected in LTED cells compared to that in MCF-7 cells. Additionally, siTUBB2B restores 2-MeO-E2-mediated inhibition of LTED cell proliferation. Other microtubule-targeting agents, i.e., paclitaxel, nocodazole, and colchicine, were not selective for LTED cells. Therefore, 2-MeO-E2 can be an antiproliferative agent to suppress LTED cell proliferation.

Predicting the short-term prognosis of patients with terminal cancer is important for treatment decisions and improving patients’ quality of life. Recently, it has been reported that the Barthel Index (BI) can predict short-term prognosis. This study aimed to distinguish the BI items that can more accurately predict the short-term prognosis of terminal cancer patients from among the other BI items. This study compared the accuracy of predicting the 1-, 2-, and 3-week prognosis of BI and individual BI items in 158 cancer patients who died between January 2018 and June 2020 at the Onomichi Municipal Hospital in Japan. For predicting the 1- and 2-week prognosis, the BI item “feeding” scores of 0/5 and 10 showed higher accuracies (0.766 and 0.715, respectively) than BI scores between 0–15/20–100. For predicting a 3-week prognosis, the BI item “mobility” scores of 0, 5/10, 15 and the BI item “grooming” scores between 0/5 showed higher accuracies (0.627 and 0.614, respectively) than BI scores between 0–35/40–100. BI and individual BI items may be an option for prognostic prediction in terminal cancer patients.

Detailed in vitro studies on the effects of perfluorooctanoic acid (PFOA) have demonstrated that activation of peroxisome proliferator-activated receptor α (PPARα) is a key process by which PFOA affects the malignancy of estrogen receptor α (ERα)-positive breast cancer cells. However, there is very little information on the PPARα-regulated genes responsible for the effects of PFOA in ERα-negative breast cancer cell malignancy. We recently demonstrated that fatty acid 2-hydroxylase (FA2H) stimulates the migration of ERα-negative human MDA-MB-231 cells, and PPARα is a key factor for the induction of FA2H in these cells. However, evidence for the relationship between PFOA exposure and PPARα-FA2H axis-driven migration has not been obtained. Here we analyzed the effects of PFOA on PPARα transcription and FA2H expression in relation to MDA-MB-231 cell migration. We found that simultaneously with stimulated migration, PFOA upregulated FA2H and activated the transcription of PPARα. FA2H-selective siRNA, but not siRNA control, clearly dampened PFOA-mediated cell migration. There is an inhibitory interaction between PPARα and PPARβ/δ (i.e., PPARβ/δ can suppress PPARα-mediated transcription) in MDA-MB-231 cells, but even in the presence of PPARβ/δ expression, PFOA appeared to free PPARα to upregulate FA2H. Collectively, our findings show that i) PFOA activates PPARα-mediated transcription, ii) PFOA stimulates migration dependent on FA2H expression, and iii) mechanistically, PFOA relieves PPARβ/δ suppression of PPARα activity to upregulate FA2H in MDA-MB-231 cells.

Here, we established an ethanol extraction method and obtained extracts of Neopyropia yezoensis cultivated in three different locations (extracts A-C) in the Seto Inland Sea (Setonaikai). The effects of the extracts on 10 human cancer cells derived from seven different organs were investigated. Extract A exerted the strongest anti-proliferative effects on all types of cancer cells, including an endocrine therapy-resistant aggressive breast cancer model, LTED cells. We analyzed the effects of the extracts on MCF-7 (parental cells for producing LTED cells)/LTED cells, along with four established anti-proliferative agents (etoposide, LY2835219, paclitaxel, and trichostatin A) with different action mechanisms. The inhibitory effects of extract A on both breast cancer cells were comparable with those of paclitaxel, although the other agents showed a preferable reduction in MCF-7 cell viability. We provide evidence of the involvement of component(s), especially those of extract A of N. yezoensis, which exerted anti-proliferative effects on cancer cells.

Cadmium (Cd) is recognized as a highly toxic heavy metal for humans in part because it is a multi-organ carcinogen. To clarify the mechanism of Cd carcinogenicity, we have established an experimental system using rat liver TRL1215 cells exposed to 2.5 μM Cd for 10 weeks and then cultured in Cd-free medium for an additional 4 weeks (total 14 weeks). Recently, we demonstrated, by using this experimental system, that 1) Cd stimulates cell invasion by suppression of apolipoprotein E (ApoE) expression, and 2) Cd induces DNA hypermethylation of the regulatory region of the ApoE gene. However, the underlying mechanism(s) as well as other potential genetic participants in the Cd-stimulated invasion are undefined. In the present work, we found that concurrent with enhanced invasion, Cd induced oxidative stress, coupled with the production of oxidative stress-sensitive metallothionein 2A (MT2A), which lead to down-modulation of ten-eleven translocation methylcytosine dioxygenase 1 (TET1: DNA demethylation) in addition to ApoE, without impacting DNA methyltransferases (DNMTs: DNA methylation) levels. Furthermore, the expression of tissue inhibitor of metalloproteinase 2 and 3 (TIMP2 and TIMP3) that are positively regulated by TET1, were decreased by Cd. The genes (ApoE/TET1/TIMP2/TIMP3) suppressed by Cd were further suppressed by hydroquinone (HQ; a reactive oxygen species [ROS] producer), whereas N-acetyl-l-cysteine (NAC; a ROS scavenger) prevented the suppression of their expression by HQ. In addition, NAC reversed their expression suppressed by Cd. Cd-stimulated cell invasion was clearly dampened by NAC in a concentration-dependent manner. Overall these findings suggest that 1) altered TET1 expression and activity together with ApoE are likely involved in the enhanced invasiveness due to Cd exposure, and 2) Cd down-regulation of TET1 likely evokes a reduction in ApoE expression (possible by DNA hypermethylation), and 3) anti-oxidants are effective in abrogation of the enhanced invasiveness that occurs concurrently with Cd-induced malignant transformation.

The association between the latent preliminary group of locomotive syndrome (LS) and handgrip strength was investigated among 96 women over age 40 years who attended health promotion events that were held 5 times between Sep. 2016 and Nov. 2019. The latent preliminary group of LS was evaluated based on three factors: bone mineral density, muscle mass and toe grip strength as an evaluation of the balance ability. The ratios of participants that declined in each factor were 82.3％, 36.5％ and 26.0％, respectively. There was a significant relationship between handgrip strength and bone mineral density, muscle mass and toe grip strength. Participants were divided into two groups based on the number of factors since it is accompanied by the functional decline of the plural locomotive organs for the onset of LS. Participants with zero or one factor were categorized as the “non-pre-LS group”. Participants with two or three factors were categorized as the “latent pre-LS group”. There was a significant difference between the median value of handgrip strength between the non-pre-LS group and the latent pre-LS group. ROC analysis indicated the cut-off value of handgrip strength corresponding to latent pre-LS was 26.0 kg, which was 6.13 as the odds ratio estimated by logistic regression analysis. The measurement of handgrip strength might be a convenient screening tool for latent pre-LS in the community pharmacy.

Here, we established an ethanol extraction method and obtained extracts of Neopyropia yezoensis cultivated in three different locations (extracts A-C) in the Seto Inland Sea (Setonaikai). The effects of the extracts on 10 human cancer cells derived from seven different organs were investigated. Extract A exerted the strongest anti-proliferative effects on all types of cancer cells, including an endocrine therapy-resistant aggressive breast cancer model, LTED cells. We analyzed the effects of the extracts on MCF-7 (parental cells for producing LTED cells)/LTED cells, along with four established anti-proliferative agents (etoposide, LY2835219, paclitaxel, and trichostatin A) with different action mechanisms. The inhibitory effects of extract A on both breast cancer cells were comparable with those of paclitaxel, although the other agents showed a preferable reduction in MCF-7 cell viability. We provide evidence of the involvement of component(s), especially those of extract A of N. yezoensis, which exerted anti-proliferative effects on cancer cells.

Bisphenol A (BPA) has been shown to induce the activation of nuclear estrogen receptor α/β (ERα/β) in both in vitro and in vivo settings. We originally obtained a 4-methyl-2,4-bis(4-hydroxyphenyl)pent-1-ene (MBP), a possible active metabolite of BPA, strongly activating the ERs-mediated transcription in MCF-7 cells with an EC50 of 2.8 nM (i.e., BPA's EC50 = 519 nM). Environmental estrogens can also target G protein-coupled estrogen receptor 1 (GPER1), a membrane-type ER. However, the effects of BPA/MBP on GPER1, have not yet been fully resolved. In this study, we used MCF-7, a ERα/ERβ/GPER1-positive human breast cancer cell line, as a model to investigate the effects of the exposure to BPA or MBP. Our results revealed that at concentrations below 1 nM MBP, but not BPA, downregulates the expression of GPER1 mRNA via upregulated ERβ, and the MCF-7 cells pre-treated with MBP display resistance to GPER1 agonist G-1-mediated anti-proliferative effects. Because GPER1 can act as a tumor suppressor in several types of cancer including breast cancer, the importance of MBP-mediated decrease in GPER1 expression in breast cancer cells is discussed.

For the estimation of short-term prognosis in terminal cancer patients, it is important to establish a prognostic index that does not involve blood tests. We compared the prognostic ability of the Barthel Index (BI) with the Glasgow Prognostic Score (GPS). Ninety-seven inpatients with terminal cancer at Onomichi Municipal Hospital who died between 2018 and 2019 were retrospectively analyzed. The sensitivity, specificity, accuracy, and area under the receiver operating characteristic curve (AUROC) were compared between the BI and GPS. For predicting the 15 day prognosis, the BI showed higher specificity, accuracy, and AUROC than the GPS. For predicting the 30 day prognosis, the BI showed higher sensitivity, accuracy, and AUROC than the GPS. The BI can predict the 15 or 30 day prognosis in terminal cancer patients. As the BI does not require blood tests, it may be an option for prognostic prediction in terminal cancer patients.

The functional role of fatty acid 2-hydroxylase (FA2H) is controversial in the field of cancer biology due to the dual role of FA2H, particularly related to its interaction with triple-negative breast cancer (TNBC). A previous biochemical- and clinical-focused study suggested that FA2H could dampen TNBC aggressiveness. However, another epidemiological study demonstrated that FA2H expression is associated with shorter disease-free survival in TNBC cases. We reported that FA2H is a peroxisome proliferator-activated receptor α (PPARα)-regulated gene in human breast cancer MDA-MB-231 cells, in vitro experimental models for TNBC analysis. PPARα activation by its ligand reportedly results in an aggressive MDA-MB-231 cell phenotype, as well as estrogen receptor α (ERα)-positive MCF-7 cells. The results of this study show that i) MDA-MB-231 cells express very low levels of FA2H compared to the MCF-7 cells, reflecting a low basal-level PPARα-driven transcriptional activity compared to the MCF-7 cells, and ii) the increased FA2H expression stimulates the MDA-MB-231 and MCF-7 breast cancer cell migration without affecting proliferation. Taken together, our findings indicate that FA2H might be a breast cancer cell migration stimulator, independently of the ERα expression status.

Prognostic prediction has been reported to affect the decision of doctors and non-physician health care providers such as nurses, social workers, pastors, and hospice volunteers on the selection of appropriate medical interventions. This was a case of a 65-year-old woman who presented with a poor oral intake. The patient had a history of sigmoid colon cancer with abdominal wall metastasis and peritoneal dissemination. On the day of admission, nausea, anorexia, and malaise were noted, requiring immediate intervention. The patient’s prognosis was predicted using the Palliative Prognostic Index. The pharmacist suggested the use of dexamethasone tablets in order to alleviate the patient’s symptoms. Indeed, the administration of dexamethasone alleviated the symptoms of nausea, loss of appetite, and malaise. To the best of our knowledge, this is the first case report to demonstrate that prognosis prediction is important not only for other medical staff but also for pharmacists when deciding the need to initiate a treatment and continue such treatment, and when providing pharmacist interventions.

Assessment of activities of daily living (ADL) is used in the prognosis of terminal cancer patients. The possibility of a relationship between the Barthel index (BI), which is an index of ADLs, and mortality among terminal cancer patients was indicated in the previous study. Using a retrospective analysis of electronic medical records of terminal cancer patients, it was determined which components of the BI had influence on mortality in this study. This survey was comprised of 126 patients who died of cancer from January 2016 to December 2017. The mortality rate determined on the 30th day post-hospitalization was 77.8 ％ . The relationships between the mortality rate and the BI, and between the mortality rate and individual components of the BI, were analyzed using a multivariate analysis, the Cox proportional hazards model. The median age (min – max) was 77.5 (30 – 96) years, and 65.9 ％ of the sub-jects were male. In the group that required total or partial bathing assistance, there was no significant difference in the mortality rate compared to the groups that were able to independently bathe themselves, but significant differences in the mortality rates of other items were recognized among those groups. These results indicate that involvement in the prognosis of terminal cancer patients might be different among each item of BI.

Both cholesterol and α-tocopherol are essential lipophilic nutrients for humans and animals. Although cholesterol in excess causes severe problems such as coronary heart disease, it is a necessary component of cell membranes and is the precursor for the biosynthesis of steroid hormones and bile acids. Niemann–Pick C1-like 1 (NPC1L1) is a cholesterol transporter that is highly expressed in the small intestine and liver in humans and plays an important role in cholesterol homeostasis. Cholesterol promotes NPC1L1 endocytosis, which is an early step in cholesterol uptake. Furthermore, α-tocopherol is the most active form of vitamin E, and sufficient amounts of vitamin E are critical for health. It has been reported that NPC1L1 mediates α-tocopherol absorption; however, the mechanisms underlying this process are unknown. In this study, we found that treatment of cells that stably express NPC1L1-GFP with α-tocopherol promotes NPC1L1 endocytosis, and the NPC1L1 inhibitor, ezetimibe, efficiently prevents the α-tocopherol-induced endocytosis of NPC1L1. Cholesterol binding to the N-terminal domain (NTD) of NPC1L1 (NPC1L1-NTD) is essential for NPC1L1-mediated cholesterol absorption. We found that α-tocopherol competitively binds NPC1L1-NTD with cholesterol. Furthermore, when cells stably expressed NPC1L1ΔNTD-GFP, α-tocopherol could not induce the endocytosis of NPC1L1ΔNTD. Taken together, these results demonstrate that NPC1L1 recognizes α-tocopherol via its NTD and mediates α-tocopherol uptake through the same mechanism as cholesterol absorption.

The ability of catechins and their related compounds to inhibit breast cancer resistance protein (BCRP) function in Caco-2 cell monolayers was investigated with mitoxantrone as a BCRP substrate. The gallate or pyrogallol moiety on the catechin structure seemed to promote increased cellular accumulation and inhibit efflux transport of mitoxantrone. The ability of gallate catechins such as (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG) to increase cellular accumulation and inhibit efflux transport of mitoxantrone was greater than that of nongallate catechins. Gallic acid octyl ester (GAO) also increased intracellular mitoxantrone accumulation. Experiments using GAO derivatives indicated that the gallate moiety required the presence of a long carbon chain for BCRP inhibition. Cellular accumulation and reduced efflux transport of mitoxantrone were greater with epigallocatechin 3-(3″-O-butyl) gallate than with EGCG. EGCG inhibition of BCRP seemed to be restricted by hydrophobicity. The co-administration of catechins, particularly EGCG and related compounds, with greater hydrophobicity may increase the therapeutic activities of BCRP substrates such as mitoxantrone.

The influence of the opioid oral rescue dose (RD) management method on opioids usage was investigated using electronic medical records in patients receiving opioid treatment from December 2014 to May 2015.