A Synthetic Chenodeoxycholic Acid Derivative, HS-1200-induced Apoptosis of RBL-2H3 Cells / 대한해부학회지

Bile acids and synthetic bile acid derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. Although synthetic chenodeoxycholic acid (CDCA) derivatives have been demonstrated to induce apoptosis of various cancer cells, there is no report on their effect on RBL-2H3 basophilic leukemia cell line to date. Therefore, this study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in RBL-2H3 cells treated with a synthetic CDCA derivative, HS-1200. The viability and the growth inhibition of RBL-2H3 cells were assessed by MTT assay and clonogenic assay respectively. The Hoechst staining and DNA electrophoresis were conducted to observe RBL-2H3 cells undergoing apoptosis. RBL-2H3 cells were treated with HS-1200, and Western blotting, immunocytochemistry, confocal microscopy, DNA hypoploidy assay, MMP activity and proteasome activity were performed. HS-1200 treatment of RBL-2H3 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Furthermore, HS-1200 treatment result in the alteration of G1 cell cycle-related proteins. And tested RBL-2H3 cells showed several lines of apoptotic manifestation.We presented data indicating that HS-1200 induces apoptois via the proteasome, mitochondria and caspase pathway, and induces the alteration of the G1 cell cycle-related proteins in RBL-2H3 cells. Therefore our data provide the possibility that HS-1200 could be as a novel therapeutic strategy in the allergy treatment.

Apoptotic Effect of Co-treatment with Chios Gum Mastic and HS-1200 on G361 Human Melanoma Cell Line / 대한해부학회지

Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. Recently it reported that CGM induce apoptosis in a few cancer cells in vitro. Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of cotreatment with a natural product, CGM and a CDCA derivative, HS-1200 on G361 human melanoma cells. To investigate whether the co-treatment of CGM and HS-1200 compared with each single treatment efficiently reduced the viability of G361 cells, MTT assay was conducted. To investigate augmentation of apoptosis in G631 cells co-treated with CGM and HS-1200, DNA electrophoresis, Hoechst staining, proteasome activity assay, flow cytometry, Westen blot analyses, immunofluorescent staining and confocal microscopy were performed. In this study, G361 cells co-treated with CGM and HS-1200 showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, activation of caspase-9, caspase-3, PARP and DFF45 (ICAD), and up-regulation of Bax whereas each single treated G361 cells did not. Although the single treatment of 40 micro/mL CGM or 25 micro HS-1200 for 24 hrs did not induce apoptosis, the co-treatment of them induced prominently apoptosis. Therefore, combination therapy of CGM and HS-1200 could be considered, in the future, as an alternative therapeutic strategy for human melanoma.

Modulation of Apoptosis and Cell Cycle by Synthetic Bile Acid Derivatives / 대한해부학회지

This paper outlines the current understanding of cell cycle modulation and induction of apoptosis in cancer cells by natural and synthetic bile acid. Bile acid homeostasis is tightly regulated in health, and the cellular and tissue concentrations of bile are restricted. However, when pathophysiological processes impair biliary secretion, hepatocytes are exposed to an elevated concentration of bile acids, which triggers cell death. In this context, we have synthesized several new bile acid derivatives. These synthetic bile acids modulate the cell cycle and induce apoptosis in several human cancer cells similar to the effects of natural bile acids. In human breast and prostate cancer cells with different tumor suppressor p53 status, synthetic bile acid induced growth inhibition and apoptosis, and these changes were associated with upregulation of Bax and p21WAF1/CIP1 through a p53-independent pathway. In Jurkat human T cell leukemia cells, the synthetic bile acids induced apoptosis through caspase activation. The synthetic bile acids induced apoptosis in a JNK-dependent manner in SiHa human cervical cancer cells through the induction of Bax and activation of caspases in PC3 prostate cancer cells and induction of G1 phase arrest of the cell cycle in HT29 colon cancer cells. The synthetic bile acids also induced apoptosis in four human glioblastoma multiform cell lines (e.g., U-118MG, U-87MG, T98G, and U-373MG) and one human TE671 medulloblastoma cell line. A chenodeoxycholic acid derivative, called HS-1200, significantly decreased the growth of TE671 medulloblastoma tumor size and increased lifespan in nonobese diabetic and severe combined immunodeficient (NOD/SCID) mice. These findings suggest that these new synthetic bile acids, which are novel apoptosis mediators, might be applicable to the treatment of various human cancer cells.

Synthetic Bile Acid Derivative HS-1200-induced Apoptosis of Human Osteosarcoma Cells / 대한해부학회지

Bile acids and synthetic its derivatives induced apoptosis in various kinds of cancer cells and had anticancer effects. However, it wasn`t discovered those materials have apoptosis induced effects on osteosarcoma cells. The present study was done to examine the synthetic bile acid derivatives induced apoptosis on osteosarcoma cells and such these apoptosis events. The synthetic bile acid derivatives, chenodeoxycholic acid (CDCA) induced the cell death on human osteosarcoma (HOS) cells contrary to ursodeoxycholic acid (UDCA). HS-1200, a synthetic derivative of CDCAs, was chosen to experiment apoptosis events in HOS cells. HOS cells treated with HS-1200 showed nucleus condensation, cytochrom c release, Bax/Bcl-xL alteration, activation of caspase-3 and caspase-activated deoxyribonuclease (CAD), and degradation of poly (ADP-ribose) polymerase (PARP). Though this study needs more investigations, these in vitro data suggest that treatment of the synthetic bile acid derivatives can give medical therapy on HOS cells.

A Novel Chenodeoxycholic Derivative HS-1200 Enhances Radiation-induced Apoptosis in Human MCF-7 Breast Cancer Cells / 대한방사선종양학회지

PURPOSE: To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. MATERIALS AND MEHTODS: Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37degrees C with 5% CO2 in air atmosphere. After removal of HS-1200, cells were irradiated with 2~8 Gy X-ray, and then cultured in drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16muM of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40muM of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken RESULTS: Treatment of MCF-7 cells with 40muM of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential (delta psi m) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. CONCLUSION: We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio in HS-1200 co-treatment group underlies the increased radiosensitivity of MCF-7 cells. Further futures studies are remained elusive.