EP4-induced mitochondrial localization and cell migration mediated by CALML6 in human oral squamous cell carcinoma.

Lymph node metastasis, primarily caused by the migration of oral squamous cell carcinoma (OSCC) cells, stands as a crucial prognostic marker. We have previously demonstrated that EP4, a subtype of the prostaglandin E2 (PGE2) receptor, orchestrates OSCC cell migration via Ca2+ signaling. The exact mechanisms by which EP4 influences cell migration through Ca2+ signaling, however, is unclear. Our study aims to clarify how EP4 controls OSCC cell migration through this pathway. We find that activating EP4 with an agonist (ONO-AE1-473) increased intracellular Ca2+ levels and the migration of human oral cancer cells (HSC-3), but not human gingival fibroblasts (HGnF). Further RNA sequencing linked EP4 to calmodulin-like protein 6 (CALML6), whose role remains undefined in OSCC. Through protein-protein interaction network analysis, a strong connection is identified between CALML6 and calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2), with EP4 activation also boosting mitochondrial function. Overexpressing EP4 in HSC-3 cells increases experimental lung metastasis in mice, whereas inhibiting CaMKK2 with STO-609 markedly lowers these metastases. This positions CaMKK2 as a potential new target for treating OSCC metastasis. Our findings highlight CALML6 as a pivotal regulator in EP4-driven mitochondrial respiration, affecting cell migration and metastasis via the CaMKK2 pathway.

Metallothioneins regulate the adipogenic differentiation of 3T3-L1 cells via the insulin signaling pathway.

Knockout of metallothionein (MT) genes contributes to a heavier body weight in early life and the potential to become obese through the intake of a high fat diet (HFD) in mice. It has thus been suggested that MT genes regulate the formation of adipose tissue, which would become the base for later HFD-induced obesity. We evaluated the fat pads of mice during the lactation stage. The fat mass and adipocyte size of MT1 and MT2 knockout mice were greater than those of wild type mice. Next, we assayed the ability of small interfering RNA (siRNA) to silence MT genes in the 3T3-L1 cell line. The expressions of MT1 and MT2 genes were transiently upregulated during adipocyte differentiation, and the siRNA pretreatment led to the suppression of the expression of both MT mRNAs and proteins. The MT siRNA promoted lipid accumulation in adipocytes and caused proliferation of post-confluent preadipocytes; these effects were suppressed by an inhibitor of phosphatidylinositol 3-kinase (LY294002). In addition, MT siRNA promoted insulin-stimulated phosphorylation of Akt, a downstream kinase of the insulin signaling pathway. Enhanced lipid accumulation in 3T3-L1 cells resulting from MT-gene silencing was inhibited by pretreatment with an antioxidant, N-acetylcysteine, used as a substitute for antioxidant protein MTs. These results suggest that interference in MT expression enhanced the activation of the insulin signaling pathway, resulting in higher lipid accumulation in 3T3-L1 adipocytes.

Change of fatty acid composition of the lumbar longissimus during the final stage of fattening in the Japanese Black cattle.

Consideration of the shortened fattening period seems to be worthwhile for the realization of profitable beef production. In this study, change of fatty acid composition of the lumbar longissimus during the final stage of fattening was investigated in Japanese Black cattle. Each of 110 fattening animals was sampled three times: the initial two samples were taken by biopsy (25.7 months and 27.5 months on average) and the final one was from carcasses (29.9 months on average). Preliminary analysis indicated that removing muscle tissues from the constant body position of the living animals should be essential for sampling. Average monounsaturated fatty acids (MUFA) at three sampling points were 58.1%, 58.5% and 60.5%, and the differences of the third sampling with the first and second samplings were significant. Both in steers and heifers, MUFA also increased as the fattening stage proceeded, and MUFA of the heifers at all the sampling points were significantly higher than those of the steers. The increasing rate of MUFA rose from 0.21 percentage points (pp)/month at period 1 (from the first sampling to the second sampling) to 0.84 pp/month at period 2 (from the second sampling to the slaughter).

Deficiency of metallothionein-1 and -2 genes shortens the lifespan of the 129/Sv mouse strain.

Metallothionein (MT) family proteins are small molecular weight and cysteine-rich proteins that regulate zinc homeostasis and have potential protective effects against oxidative stress and toxic metals. To investigate whether MTs play a role in longevity determination in mammals, we measured the lifespans of wild-type (WT) and MT-1 and -2 gene knockout (MTKO) mice in a 129/Sv genetic background. MTKO mice of both sexes had shorter lifespans than WT mice. In particular, male MTKO mice living beyond the mean lifespan exhibited signs of weight loss, hunchbacked spines, lackluster fur and an absence of vigor. These results suggest that lifespan is shortened due to accelerated senescence in the absence of MT genes.