Hailan Li

Indole-3-carbinol inhibits prostate cancer cell migration via degradation of beta-catenin.

We determined whether indole-3-carbinol (I3C) could affect DU145 human prostate carcinoma cell migration to prevent the development and progression of prostate cancer. Although previous studies have shown anticancer properties of I3C in various cancer cell lines, it has not been determined how I3C regulates epidermal growth factor (EGF)-induced migration and related signaling pathways. DU145 cells were treated with I3C (100 microM) in the absence or presence of EGF (10 ng/ml). Our results showed that I3C significantly inhibited DU145 cell migration with and without EGF stimulation. It has been reported that the beta-catenin signaling pathway controls androgen receptor (AR)-mediated prostate cancer progression, which plays a key role in the metastasis of prostate cancer. Western blot analysis demonstrated that I3C led to the phosphorylation of beta-catenin and subsequent degradation of beta-catenin in the absence and presence of EGF. In contrast, I3C did not have any effect on the expression of beta-catenin mRNA. From these results, we suggest that I3C inhibits EGF (dependent or independent)-induced DU145 cell migration through beta-catenin degradation.

Phosphatidylcholine induces apoptosis of 3T3-L1 adipocytes

BackgroundPhosphatidylcholine (PPC) formulation is used for lipolytic injection, even though its mechanism of action is not well understood.MethodsThe viability of 3T3-L1 pre-adipocytes and differentiated 3T3-L1 cells was measured after treatment of PPC alone, its vehicle sodium deoxycholate (SD), and a PPC formulation. Western blot analysis was performed to examine PPC-induced signaling pathways.ResultsPPC, SD, and PPC formulation significantly decreased 3T3-L1 cell viability in a concentration-dependent manner. PPC alone was not cytotoxic to CCD-25Sk human fibroblasts at concentrations <1 mg/ml, whereas SD and PPC formulation were cytotoxic. Western blot analysis demonstrated that PPC alone led to the phosphorylation of the stress signaling proteins, such as p38 mitogen-activated protein kinase and c-Jun N-terminal kinase, and activated caspase-9, -8, -3 as well as cleavage of poly(ADP-ribose) polymerase. However, SD did not activate the apoptotic pathways. Instead, SD and PPC formulation induced cell membrane lysis, which may lead to necrosis of cells.ConclusionsPPC results in apoptosis of 3T3-L1 cells.

Effects of Cervi cornus Colla (deer antler glue) in the reconstruction of a skin equivalent model

The aim of this study was to investigate the effects of Cervi cornus Colla (CCC) in the reconstruction of skin equivalent (SE). H&E staining showed that SE containing hyaluronic acid (HA) or HA and CCC had a thicker epidermis than the control SE. Immunohistochemical staining showed that p63 was mainly present at the basal layer of the epidermis in the HA and CCC model. Involucrin was obviously expressed in the upper layer of the epidermis in the HA and CCC model. Moreover, we observed that integrins α6 and β1 were strongly expressed along the basement membrane zone in the HA and CCC model, in which the dermis expressing type I collagen was more compact. In conclusion, our data indicate that CCC contributed to the formation of epidermis, basement membrane, and extracellular matrix in the reconstruction of SE and suggest that CCC may be a useful adjuvant in the reconstruction of SE.

Photo-activated 5-hydroxyindole-3-acetic acid induces apoptosis of prostate and bladder cancer cells

5-Hydroxyindole-3-acetic acid (5-HIAA), an indole derivative, is the main metabolite of serotonin in the human body. We determined whether or not ultraviolet B (UVB)-activated 5-HIAA (5-HIAA(UVB)) affects the viability of human prostate (LnCaP and PC-3) and bladder cancer cells (TCCSUP). While 5-HIAA alone had no cytotoxic effect at <1mM, 5-HIAA(UVB) induced LnCaP, PC-3, and TCCSUP cell death in a time- and dose-dependent manner. Cell cycle analysis showed that 5-HIAA(UVB) markedly increased the sub-G(0)/G(1) phase and resulted in cell cycle disruption. To elucidate the death mechanism by 5-HIAA(UVB), we examined the signal transduction pathways related to apoptosis using Western blot analysis. 5-HIAA(UVB) led to phosphorylation of stress-activated signaling proteins, such as c-Jun N-terminal kinase (JNK) and/or p38 mitogen-activated protein kinase (MAPK). Furthermore, 5-HIAA(UVB) activated caspase-8, -9, and -3 and cleaved poly(ADP-ribose) polymerase (PARP), which are indicators of apoptosis. From these findings, the present study demonstrated that 5-HIAA(UVB) induces apoptotic cell death of prostate and bladder cancer cells via stress-mediated signaling and apoptotic pathways. Therefore, we suggest that 5-HIAA might be used as a new photosensitizer for photodynamic cancer therapy.

KHG26792 Inhibits Melanin Synthesis in Mel-Ab Cells and a Skin Equivalent Model

The purpose of this study is to characterize the effects of KHG26792 (3-(naphthalen-2-yl(propoxy) methyl)azetidine hydrochloride), a potential skin whitening agent, on melanin synthesis and identify the underlying mechanism of action. Our data showed that KHG26792 significantly reduced melanin synthesis in a dose-dependent manner. Additionally, KHG26792 downregulated microphthalmia-associated transcription factor (MITF) and tyrosinase, the rate-limiting enzyme in melanogenesis, although tyrosinase was not inhibited directly. KHG26792 activated extracellular signal-regulated kinase (ERK), whereas an ERK pathway inhibitor, PD98059, rescued KHG26792-induced hypopigmentation. These results suggest that KHG26792 decreases melanin production via ERK activation. Moreover, the hypopigmentary effects of KHG26792 were confirmed in a pigmented skin equivalent model using Cervi cornus Colla (deer antler glue), in which the color of the pigmented artificial skin became lighter after treatment with KHG26792. In summary, our findings suggest that KHG26792 is a novel skin whitening agent.

Fucoidan promotes the reconstruction of skin equivalents.

In this study we investigated the effects of fucoidan on the proliferation of fibroblasts and the reconstruction of a skin equivalent (SE). Fucoidan significantly stimulated the proliferation of CCD-25Sk human fibroblasts and Western blot analysis demonstrated that fucoidan markedly increased the expression of cyclin D1 and decreased the expression of p27. Fucoidan was used to reconstruct SE. Immunohistochemical staining showed that the addition of fucoidan to dermal equivalents increased expression of proliferating cell nuclear antigen (PCNA) and p63. In addition, expression of α6-integrin was significantly increased by fucoidan, whereas expression of β1-integrin, type 1 collagen, elastin, fibronectin did not markedly change. These results suggest that fucoidan has positive effects on epidermal reconstruction and will therefore be beneficial in the reconstruction of SE.

Avian Collagen Is Useful for the Construction of Skin Equivalents

As a result of restrictions on animal experimentation, improved skin equivalents (SEs) are needed as alternative test models. This work investigated the effects of avian collagen on the construction of SEs, and to the best of our knowledge is the first study to do so. Hematoxylin and eosin and immunohistochemical staining were used to analyze the SEs. In models containing avian collagen as a dermal equivalent (DE) ingredient, fibroblast proliferation increased by about 60% relative to the control model. Immunohistochemical staining showed that the expression of proliferating cell nuclear antigen (PCNA) and p63 increased in the avian collagen models, while the expression of involucrin, integrin α6, and integrin β1 remained unchanged. Next, DEs were cryopreserved to allow the easier creation of SEs. Keratinocytes were seeded on thawed DEs, and SEs were constructed. Avian collagen increased the viability of DEs relative to the control. Furthermore, avian collagen increased the expression of PCNA and p63 in keratinocytes on thawed DEs. The results indicate that DEs containing avian collagen can be thawed as needed after cryopreservation. Avian collagen can improve the construction of SEs and be used as part of a dermal kit for SE construction.

Imidazole inhibits B16 melanoma cell migration via degradation of β‐catenin

Objectives In the present study, we determined whether or not imidazole affects B16 murine melanoma cell migration to prevent melanoma metastasis.

Assessment of Skin Toxicity Using Skin Equivalents Containing Cervi cornus Colla

To substitute animal test, skin equivalents (SEs) have been developed for skin irritation and corrosion test. Recently, we have developed new SEs containing Cervi cornus Colla (CCC). In the present study, we used the SEs for cutaneous cytotoxicity test. Sodium dodecylsulfate (SDS) or sodium carbonate was applied to the SEs-, and the epidermal damage by H&E and immunohistochemical stains was evaluated. Our results showed that SDS or sodium carbonate affected the epidermal part of SEs containing CCC in a dose-dependent manner and decreased the expression of p63. It is concluded that SEs containing CCC could be used for an alternative model of animal test and would be greatly helpful in the development of in vitro irritation and corrosion test.