Altered maturation of dendritic cells by taxol, an anticancer drug

주홍구 대한수의학회 2003 Journal of Veterinary Science Vol.4 No.3

Granulocyte-macrophage colony stimulating factor protects dendritic cells from anticancer drug-induced apoptosis

주홍구,Joo, Hong-Gu The Korean Society of Veterinary Science 2003 大韓獸醫學會誌 Vol.43 No.4

Dendritic cells (DCs) play an essential role in a variety of immune reactions involving $CD4^+$ T cells and have been used to enhance tumor-specific immune responses. Immunosuppression in patients with cancer includes the downregulation of function and number of DCs. Although DCs have been studied, the apoptosis of Des induced by anticancer drugs for chemotherapy remains largely uncharacterized. This study demonstrated that GM-CSF protects DCs from 5-fluorouracil (5-FU) or mitomycin C-induced apoptosis. After 6 - 10 days culture, DCs were characterized by specific surface marker, CD11c and MHC class II. MTT assay revealed that GM-CSF significantly enhanced the viability of DCs treated with 5-FU or mitomycin C. The percentage of dead cells of DCs was determined by cell size using FACScan and GM-CSF was clearly effective. However, GM-CSF did not increase the expression of MHC class II on viable DCs gated, suggesting that GM-CSF may differentially regulate critical factors involved in the function of DCs. For the quantitative analysis of apoptosis, annexin V-FITC staining was performed. 5-FU induced the apoptosis of DCs and GM-CSF significantly protects DCs from 5-FU-induced apoptosis. Taken together, the results in this study that GM-CSF has an anti-apoptosis effect on DCs may provide patients with cancer with clinical benefits to overcome the immunosuppression induced by the decrease of number and functional insufficiency of DCs.

위암에서 Galectin-3 단백질 발현에 관한 연구

주홍구,우희종 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

The expression of galectin-3 protein in four different gastric cancer cell lines and lymph node tissues of patients with gastric carcinoma was studied by Western blotting, immundhistochenustry and flow cytometry analysis. Western blot analysis of galectin-3 using anti-galectin-3 , mAb indicated that a 29 kDa protein was expressed in 3 gastric cancer cell lines, AGS, KATO BI , and SNU16, but not in SNU1 which was a non-adherent cell line. Interestingly, SNU16 cells secreted galectin-3 into the culture media. Flow cytometry analysis revealed that NIH3T3, AGS, and KATO III cells expressed the galectin-3 on the cell surface. The level a))' galectin-3 expression was highest in AGS (91.5%), high in KATO M (69.5%), and low in NIH3T3 (55.8%). The gastric cancer cells metastasized into .lymph node tissues were heavily stained whereas normal lymphocytes were not stained. This study suggested that the expression of galectin-3 and the metastasis of gastric cancer cells might be correlated.

Radioprotective activity and stimulatory effects of an acidic polysaccharide of Panax ginseng on bone marrow and immune cells

Hong-Gu Joo(주홍구) 고려인삼학회 2011 고려인삼학회 학술대회 Vol.2011 No.4

Evaluation of the effects of disulfiram, an alcohol-aversive agent with anti-cancer activity, on mouse bone marrow cells

박서로,주홍구 대한약리학회 2022 The Korean Journal of Physiology & Pharmacology Vol.26 No.3

Disulfiram (DSF) is an aldehyde dehydrogenase inhibitor. DSF has potent anti-cancer activity for solid and hematological malignancies. Although the effects on cancer cells have been proven, there have been few studies on DSF toxicity in bone marrow cells (BMs). DSF reduces the metabolic activity and the mitochondrial membrane potential of BMs. In subset analyses, we confirmed that DSF does not affect the proportion of BMs. In addition, DSF significantly impaired the metabolic activity and differentiation of BMs treated with granulocyte macrophage-colony stimulating factor, an essential growth and differentiation factor for BMs. To measure DSF toxicity in BMs in vivo, mice were injected with 50 mg/kg, a dose used for anti-cancer effects. DSF did not significantly induce BM toxicity in mice and may be tolerated by antioxidant defense mechanisms. This is the first study on the effects of DSF on BMs in vitro and in vivo. DSF has been widely studied as an anti-cancer drug candidate, and many anti-cancer drugs lead to myelosuppression. In this regard, this study can provide useful information to basic science and clinical researchers.

Cytotoxicity of fucoidan on spleen cells via the induction of early apoptosis

양혜열,주홍구 한국예방수의학회 2016 예방수의학회지 Vol.40 No.1

Fucoidan is a sulfated polysaccharide, purified from brown algae. It has multiple biological activities including anti-cancer and anti-inflammatory effects. Our previous reports demonstrated that fucoidan can stimulate spleen cells and especially high molecular weight fucoidan is responsible for the immunostimulatory activity. However, we recently found that the activity of fucoidan can be dependent on its individual batch or sources. Four different fucoidans (fucoidan A, fucoidan B, high molecular weight fucoidan, and low molecular weight fucoidan) were used for this study. MTT assay and flow cytometry analysis were performed for analysis of the activity of fucoidan. MTT assay showed that fucoidan B significantly decreased the cellular activity of spleen cells compared to fucoidan A. In addition, fucoidan B consistently killed spleen cells based on the cell size by flow cytometry analysis and the morphology by an inverted microscope. To elucidate the detailed mechanisms of cytotoxicity, fucoidan B-treated spleen cells were stained with Rhodamine 123 solution and Annexin V-FITC/propidium iodide for measurement of mitochondrial membrane potential (MMP) and early/late apoptosis, respectively. From these two assays, fucoidan B decreased the MMP and induced early apoptosis of spleen cells. Taken together, we suggest that different batches or origin of fucoidan may have differential activities on spleen cells, immunostimulatory and cytotoxic activity. The present study may provide some valuable information regarding use of fucoidan in the clinical area and in basic research.

마우스 비장세포에서 Ginsenoside Rp1의 세포자멸사 유도

오영균,주홍구,Oh, Young-Kyun,Joo, Hong-Gu 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.3

Ginsenoside Rp1 is one of ginseng saponins with chemotherapeutic activity. In this study, we investigated the effects of Rp1 on spleen cells. Spleen is a major immune organ consisted of crucial immune cells, such as T lymphocytes, B lymphocytes, natural killer cells, and some antigen-presenting cells. Although the anti-tumor potential of Rp1 was studied, the effects of Rp1 on immune cells have not investigated yet. A viability assay using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), flow cytometric analysis, Western blot analysis were used to detect cellular changes on Rp1-treated spleen cells. MTT assay showed that Rp1 decreased the viability of spleen cells. To further investigate the effects of Rp1 on activated spleen cells, we treated lipopolysaccharide (LPS) as a representative inflammatory agent and Rp1 on spleen cells in a combination. The surface expression levels of activation markers for lymphocytes, CD25 and CD69 were measured. Apoptotic analysis revealed the cytotoxic effects of Rp1 on both na$\ddot{i}$ve and activated cells, and the expression pattern of some apoptosis-related proteins was correlated to apoptotic events of cells. Taken together, ginsenoside Rp1 increases the cellular death of spleen cells and also inhibits the LPS-induced activation of spleen cells.

Dendritic cells resist to disulfiram-induced cytotoxicity, but reduced interleukin-12/23(p40) production

정한빈,주홍구 대한약리학회 2023 The Korean Journal of Physiology & Pharmacology Vol.27 No.5

Disulfiram (DSF), a medication for alcoholism, has recently been used as a repurposing drug owing to its anticancer effects. Despite the crucial role of dendritic cells (DCs) in immune homeostasis and cancer therapy, the effects of DSF on the survival and function of DCs have not yet been studied. Therefore, we treated bone marrow-derived DCs with DSF and lipopolysaccharide (LPS) and performed various analyses. DCs are resistant to DSF and less cytotoxic than bone marrow cells and spleen cells. The viability and metabolic activity of DCs hardly decreased after treatment with DSF in the absence or presence of LPS. DSF did not alter the expression of surface markers (MHC II, CD86, CD40, and CD54), antigen uptake capability, or the antigen-presenting ability of LPS-treated DCs. DSF decreased the production of interleukin (IL)-12/23 (p40), but not IL-6 or tumor necrosis factor-α, in LPS-treated DCs. We considered the granulocyte-macrophage colony-stimulating factor (GM-CSF) as a factor to make DCs resistant to DSF-induced cytotoxicity. The resistance of DCs to DSF decreased when GM-CSF was not given or its signaling was inhibited. Also, GM-CSF upregulated the expression of a transcription factor XBP-1 which is essential for DCs’ survival. This study demonstrated for the first time that DSF did not alter the function of DCs, had low cytotoxicity, and induced differential cytokine production.

마우스 수지상세포의 항원 제시 능력에 대한 이온화 규소수의 biphasic 면역조절 효과

이유정,주홍구,Lee, You-Jeong,Joo, Hong-Gu 대한수의학회 2021 大韓獸醫學會誌 Vol.61 No.2

Biosilica is a silica-based substance derived from the cell walls (frustules) of diatoms. Recently, research into biosilica's biological functions is underway, but little has been reported on the effects of biosilica on immune cells. In this study, we investigated the effect of ionized biosilica water (iBW) on dendritic cells (DCs), which play crucial roles as antigen (Ag)-presenting cells. Treatment with iBW increased the expression of immune response-related markers, closely connected to the maturation of DCs, and the production of tumor necrosis factor-alpha. In addition, iBW-treated DCs (iBW-DCs) had a lower uptake of fluorescein isothiocyanate-dextran than that of control DCs. Mixed leukocyte response analysis used for measuring the Ag-presenting capability of DCs, showed iBW-DCs had a higher capability than that of control DCs. Interestingly, DCs treated with lipopolysaccharide (LPS) and iBW had a lower level of Ag-presenting capability than that of LPS-treated DCs. Taken together, the results indicate that iBW alone can mature DCs, but it decreases the Ag-presenting capability of DCs in the presence of LPS, a representative agent of inflammation. This study may provide valuable information regarding the effect of iBW on immune cells. Further research is needed to investigate how iBW induces the observed biphasic immunomodulatory activity.

지질다당류로 활성화된 마우스 골수세포에서 구충제 Fenbendazole의 억제 효과

박서로,주홍구,Park, Seo-Ro,Joo, Hong-Gu 대한수의학회 2021 大韓獸醫學會誌 Vol.61 No.3

Fenbendazole (FBZ) is a commonly used anthelmintics in veterinary medicine that has recently been found to have anticancer effects in humans. On the other hand, few studies have examined the anti-inflammatory effects of FBZ, and its mechanism is unknown. In this study, mouse bone marrow cells (BMs) were treated with lipopolysaccharide (LPS), a representative inflammation-inducing substance, to generate a situation similar to osteomyelitis in vitro. The effect of FBZ on inflammatory BMs was examined by measuring the metabolic activity, surface marker expression, cell nuclear morphology, and mitochondrial membrane potential (MMP) of BMs. FBZ decreased the metabolic activity and MMP of LPS-treated BMs. Annexin V-fluorescein isothiocyanate/propidium iodide staining and Hoechst 33342 staining showed that FBZ reduced the number of viable cells and induced the cell death of inflammatory BMs. In addition, FBZ reduced the proportion of granulocytes more than B lymphocytes in LPS-treated BMs. Overall, FBZ induces cell death by destabilizing the MMP of LPS-induced inflammatory BMs. In addition to anthelmintic and anticancer agent, FBZ can play a role as an anti-inflammatory agent.