We have previously mapped a diet-induced hypercholesterolemia locus (Dihc2) to chromosome 14 in the F2 generation cross of high-responsive exogenous hypercholesterolemia rats and low-responsive BN rats. To identify a causal gene within this locus, we constructed intervalspecific congenic lines and carried out expression and sequencing analyses. Here we narrowed Dihc2 to a region including 33 genes and predicted transcripts and identified RGD1309450_predicted, a homologous gene of SMEK2, as a strong candidate for responsiveness to dietary cholesterol. Our finding provides new insights into the pathway underlying the individual responsiveness to dietary cholesterol in vivo. Coronary heart disease is a leading cause of mortality in most industrialized countries. Epidemiological studies support that hypercholesterolemia is a major risk factor for coronary heart disease (1). The concentration of total cholesterol in serum is a quantitative and continuous trait that is controlled by complex systems involving environmental and polygenic factors and their interactions. Dietary cholesterol is an environmental factor that raises the total concentration of serum cholesterol in humans and animals (2, 3). However, individuals vary widely in the response to dietary cholesterol, implying individual genetic variability (4). Some genes whose polymorphisms influence the response to dietary cholesterol have already been reported in humans (5). In addition, novel quantitative trait loci (QTLs) for the response to dietary cholesterol have been identified. A genetic study of the stroke-prone spontaneously hypertensive rat, which showed an exaggerated response to a high-fat, high-cholesterol diet, showed QTLs for postdietary cholesterol levels on rat chromosomes 7, 15, and 16 (6). The genetic locus for diet-dependent hypercholesterolemia in the New Zealand obese mouse was also identified on distal mouse chromosome 5 (7). QTL analyses of an intercross of CAST/Ei and DBA/2J mice fed a high-cholesterol diet identified novel a QTL for total cholesterol on mouse chromosome 9 (8). However, these novel genetic loci have not been elucidated in the identification of causal genes.Exogenously hypercholesterolemia (ExHC) rats generated from SD rats showed a 3-fold higher serum total cholesterol level than SD rats when fed a 1% cholesterol-containing diet for 2 weeks (9, 10). Thus, the ExHC rat is an appropriate model animal for evaluating the effects of dietary cholesterol on serum total cholesterol levels. To identify factors associated with the response to dietary cholesterol, we carried out QTL analyses using high-responsive ExHC rats, low-responsive BN rats, and (ExHC 3 BN)F2 progeny fed a diet containing 1% cholesterol (11). We mapped dietinduced hypercholesterolemia QTLs to rat chromosomes 5 and 14, and labeled them as diet-induced hypercholesterolemia1 (Dihc1) and Dihc2 (11).In the present study, we first constructed an Ex.BN-Dihc2 congenic strain to confirm that Dihc2 is a QTL for dietinduced hypercholesterolemia. Second, we trie...

Background: The main component of mung bean protein, accounting for more than 80%, is 8Sα globulin. Its structure closely resembles that of soybean β-conglycinin. Thereby, the mung bean protein is expected to have similar physiological effects to those of β-conglycinin, but there is no clinical evidence for these effects.Purpose of this study: The aim of this study was to confirm the positive effects of mung bean protein (GLUCODIATM) on glucose metabolism in clinical trials.Method: This clinical study was conducted using a double-blind placebo-controlled design with 45 prediabetes patients.Results: Many of the subjects were pre-diabetes with blood glucose levels exceeding 140 mg/dL by 2-hour plasma glucose level. However, the initial mean fasting plasma glucose level was less than 100 mg/dL. Therefore, mung bean protein did not lower fasting plasma glucose levels. The test period extended from summer to autumn, and increased fasting plasma glucose levels in the placebo group were observed due to seasonal factors. However, this increase was suppressed in the test group. Similarly, the mean insulin level increased in the placebo group, but the increase was also suppressed in the test group. Among obese subjects with a high body mass index, significant increases in fasting plasma glucose and insulin levels in the placebo group were observed. In the comparison between the test and the placebo groups with the average elevation value, there was a significant difference in fasting blood glucose level and significant tendencies in insulin level and homeostatic model assessment for insulin resistance value between the two groups.Conclusion: Mung bean protein suppresses fasting plasma glucose and insulin levels. Consequently, it may have an inhibitory effect on insulin resistance, a trigger of metabolic syndrome.Key words: mung bean protein, insulin, obesity, body mass index, randomized clinical trial, seasonal variation.

The receptor tyrosine kinase-like orphan receptor 1 (ROR1) regulates caveolae formation and caveolae-dependent endocytosis by interacting with caveolae components, which in turn sustains pro-survival signaling toward AKT from multiple RTKs, including EGFR, and MET. We report here a novel function of ROR1 as a scaffold for HRS and STAM1, two essential components of ESCRT-0. The present results show that ROR1 facilitates interactions of HRS and STAM1, thereby preventing the lysosomal degradation of HRS. Furthermore, interaction of ROR1 with STAM1 was found to be required to sustain binding of ROR1 to HRS as well as HRS subcellular localization. Additionally, ROR1 localized in both the limiting membrane and intraluminal vesicles (ILVs) of Rab5-induced multivesicular endosomes (MVEs) containing HRS, CD63, and EEA1 was found to regulate the formation of Rab5-induced MVEs by an association with the GTP-bound form of Rab5 in cancer cells. Notably, ROR1 depletion inhibits CD63-positive MVEs formation and reduces exosomes release. Our findings provide the first evidence that the onco-embryonic antigen receptor ROR1 regulates exosome biogenesis via MVE formation in cancer cells.

Luminal breast cancer has the highest bone metastasis frequency among all breast cancer subtypes; however, its metastatic mechanism has not been elucidated because of a lack of appropriate models. We have previously developed useful bone metastatic cell lines of luminal breast cancer using MCF7 cells. In this study, we characterized bone metastatic MCF7-BM cell lines and identified c-Jun as a novel bone metastasis marker of luminal breast cancer. The protein level of c-Jun was upregulated in MCF7-BM cells compared with that in parental cells, and its deficiency resulted in the suppression of tumor cell migration, transformation, and reduced osteolytic ability. In vivo, dominant-negative c-Jun exhibited smaller bone metastatic lesions and a lower metastatic frequency. Histological analysis revealed that c-Jun expression was heterogeneous in bone metastatic lesions, whereas c-Jun overexpression mediated a vicious cycle between MCF7-BM cells and osteoclasts by enhancing calcium-induced migration and releasing the osteoclast activator BMP5. Pharmacological inhibition of c-Jun by the Jun amino-terminal kinase (JNK) inhibitor JNK-IN-8 effectively suppressed tumorigenesis and bone metastasis in MCF7-BM cells. Furthermore, c-Jun downstream signals were specifically correlated with the clinical prognosis of patients with the luminal subtype of breast cancer. Our results illustrate the potential benefits of a therapy that targets c-Jun to prevent bone metastasis in luminal breast cancer. Implications: c-Jun expression mediates bone metastasis in luminal breast cancer by forming a vicious cycle in the bone microenvironment, which reveals potential strategies for subtype-specific bone metastasis therapy.

Luminal breast cancer has the highest bone metastasis frequency among all breast cancer subtypes, but its metastatic mechanism has not been elucidated because of the lack of appropriate metastatic cell lines. The study aim was to characterize high-osteolytic bone metastatic MCF7-BM cell lines and extract c-Jun, a novel bone metastasis marker. We found that c-Jun was upregulated in MCF7-BM cells, and its deficiency was associated with suppression of the cell migration, transformation, and stemness of BM cells. In vivo, c-Jun-deficient MCF7-TAM67 cells exhibited weaker bone metastatic ability. Additionally, c-Jun overexpression in MCF7-BM cells led to a tumor-migration promotion cycle in the bone microenvironment possibly by enhancing calcium-induced migration and releasing the osteoclast activator BMP5. Inhibition of c-Jun by JNK-IN-8, a JNK inhibitor, effectively reduced tumorigenesis activities and bone metastatic tumors. Our results indicate the potential benefits of a therapy that targets c-Jun to prevent or minimize luminal breast cancer bone metastasis.