MCF-7 세포주에서 Glutathione S-Transferase K1 (hGSTK1) 과발현에 의한 방사선 내성의 유도

김재철,신세원,Kim, Jae-Chul,Shin, Sei-One 대한방사선종양학회 2001 Radiation Oncology Journal Vol.19 No.4

목적 : 사람의 유방암 세포인 MCF-7 세포주를 대상으로 gultathione S-transferase K1 (hGSTK1) 유전자의 발현 정도 및 방사선 조사에 의한 hGSTK1 유전자의 발현 변화를 관찰하고 hGSTK1 유전자를 과발현시킴으로써 hGSTK1이 방사선 감수성에 어떤 영향을 미치는 지를 관찰하였다. 재료 및 방법 : 사람의 모유두 세포 pBluescript phagemid cDNA library로부터 선별한 hGSTK1 cDNA를 pcDNA3.1/Myc-His (+) vector에 결합시킨 후, 사람의 유방암 세포인 MCF극 세포에 이입시켰다. hGSTK1 유전자를 이입시키지 않은 MCF극 세포와 hGSTK1을 이입시킨 MCF-7 세포에 $2\~12\;Gy$의 엑스선을 조사하여 생존 분획을 비교하였다. hGSTK1 유전자를 이입시키지 않은 MCF-7 세포와 hGSTK1을 이입시킨 MCF-7 세포에서 방사선량, 분할조사 여부, 방사선 조사 후 경과 시간에 따른 hGSTK1 mRNA 발현의 차이를 보기 위하여 RT-PCR 분석을 시행하였다. 결과 : hGSTK1 유전자를 이입시키기 전의 MCF-7 세포보다 hGSTK1 유전자를 이입시킨 MCF-7 세포에서 생존 분획이 유의하게 높은 것으로 나타났다. hGSTK1 유전자를 이입시키지 않은 MCF-7 세포에서 2 Gy 생존 분획은 $0.3250{\pm}0.0319$였고, hGSTK1 유전자를 이입시킨 MCF-7 세포에서 2 Gy 생존 분획은 $0.4125{\pm}0.0325$였다 (p<0.05). 그러나 RT-PCR에 의한 hGSTK1 mRNA 분석에서는 방사선량, 분할조사 여부, 방사선 조사 후 경과 시간에 따른 발현의 차이를 볼 수 없었다. 결론 : MCF-7 세포주에서 hGSTK1의 과발현은 방사선 감수성에 영향을 미쳐서 MCF-7 세포의 생존 분획을 증가 시켰다고 볼 수 있으나 이에 대한 정확한 기전을 알기 위해서는 더 많은 연구가 필요할 것이다. Purpose : This study was conducted to assess the effects of x-irradiation on the expression of the novel glutathione S-transferase K1 gene. Materials and methods : Human glutathione S-transferase K1 (hGSTK1) DNA was purified and ligated to a pcDNA3.1/Myc-His(+) vector for the overexpression of hGSTK1 gene. MCF-7 cells were transfected with or without the recombinant hGSTK1 gene, and irradiated with 6 MV x-ray. After incubation of 14 days, cell survival was measured and compared. The expression of hGSTK1 and the effect of x-irradiation on hGSTK1 expression were also estimated in MCF-7 cells transfected with or without the hGSTK1 gene by RT-PCR. Results : Following 2 to 12 Gy of x-irradiation, the cell survivals were higher in the MCF-7 cells transfected with the hGSTK1 gene than in those without transfection. Despite the higher cell survival in the hGSTK1-transfected cells, RT-PCR for hGSTK1 mRNA revealed no significant differences according to radiation dose, fractionation, and time after irradiation. Conclusion : The MCF-7 cells transfected with the hGSTK1 gene showed higher cell survival than those without transfection of the gene. The hGSTK1 gene might be associated with the radiosensitivity of MCF-7 cell line and further analysis should be needed.

유방암 세포 주 MCF-7에서의 녹차 추출물이 p53 경로에 미치는 영향

곽인석(Inseok Kwak) 한국생명과학회 2018 생명과학회지 Vol.28 No.11

녹차(GT) 추출물의 효과를 인간 유방암 유래 세포인 MCF-7 세포를 사용하여 조사 하였다. GT추출물의 세포독성 효과를 MTT 방법을 사용하여 관찰한 결과, MCF-7 세포는 현저한 세포 독성 효과를 보였고, 이 독성 효과는 GT추출물 농도 의존적으로 증가하였다. p53과 관련 단백질인 p21/cip1과 CDK2의 연관성을 조사하기 위해 GT추출물 처리 후 웨스턴 분석법을 통해 이들 단백질의 발현을 조사하였다. GT추출물 처리 후, MCF-7 세포에서 p53단백질의 양은 농도에 따라 현저하게 증가 하였다. p21/cip1 단백질의 발현은 낮은 농도의 GT추출물에서 증가되며, 고농도에서도 감소하지 않았다. 그러나 CDK2의 단백질의 양은 높은 농도의 GT추출물에서 CDK2 발현의 급격한 감소가 관찰되었다. 이 결과는 GT추출물의 처리는 MCF-7 세포에서 p53와 p21/cip1를 증가시켜, 그 결과로 활성화 된 p21/cip1는 CDK2의 발현을 억제 함을 나타내고 있다. GT추출물이 MCF-7 세포의 세포주기에 어떤 영향을 미치는지 확인하기 위하여 FACS 분석으로 관찰한 결과, MCF-7 세포에서 세포주기의 G1 단계가 점차 증가하는 결과를 보였다. 이 결과는 GT추출물의 유방암 세포에서의 항암 효과는 세포주기의 G1 단계에서 MCF-7 세포를 정지시키는 p53에 의해 조절된다는 사실을 명확하게 보여 주고 있다. The effects of a green tea extract (GTE) were examined using the MCF-7 human breast cancer cell line. Cell viability assays using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays revealed that GTE had a significant cytotoxic effect on MCF-7 cells, depending on the concentration of GTE. Western blotting of p53 and its related proteins, p21/cip1 and CDK2, after GTE treatment revealed that a significant and concentration dependent increase in p53 protein in response to GTE. The levels of p21/cip1 proteins were also increased at low GTE concentrations were significantly increased even at the highest GTE concentrations. However, the level of CDK2 was significantly decreased by treatment with high concentrations of GTE. These results indicate that treatment with GTE increased the p53 level in MCF-7 cells, and this activation of p53 markedly elevated the levels of p21/cip1proteins, which, in turn, inhibited CDK2 expression in the MCF-7 cells. The inhibition of CDK2 expression might then affect cell cycle progression. Subsequent FACS analysis indicated that GTE treatment the gradually increased progression of the MCF-7 to the G1 phase. These results clearly demonstrate that the anti-tumor effect of GTE in MCF-7 cells is regulated by p53 arrest of the MCF-7 cells at the G1 stage of cell cycle.

Antitumor Effects of Cistanchis Herba Aqueous Extracts on MCF-7 Human Breast Cancer Cell-Xenograft Athymic Nude Mice, through Potent Immunomodulatory Activities

( Hwang Hyeon-ji ) 대구한의대학교 제한동의학술원 2022 제한동의학술원논문집 Vol.20 No.1

BACKGROUND AND PURPOSE : Cistanchis Herba (CH) are a Yang-invigorating Chinese tonic herb that is primarily used to treat kidney deficiency with symptoms such as impotence, infertility, premature ejaculation. Besides, CH extracts treatments induced significant immunomodulatory effects on 4T1, murine mammary tumor-bearing mice by increasing cytokines interleukin (IL)-2 and interferon (IFN)-γ and modulation of regulatory T-cells. Furthermore, CH treatments did not stimulate but suppressed human triple-negative MDA-MB-231 breast xenografts through their potent immunomodulatory activities. In this experiment, the antitumor, anti-cachexia and immunomodulatory effects of CH aqueous extracts (CH), which prepared and supplied by Ockchundang (Daegu, Korea), were systemically observed in MCF-7 cell xenograft athymic nude mice. METHODS : One hundred-ten 6-week-old female athymic nude mice were prepared, and MCF-7 tumor cells were implanted on the skins of right dorsal hip after 7 days acclimatization. Twenty days after tumor inoculation, eight mice in each group showing regular body weights (intact mice: 20.31 ± 1.40 g, ranged in 18.20 ~ 22.20 g; tumor bearing mice: 20.34 ± 0.95 g, ranged in 18.70 ~ 22.30 g) and tumor volumes (114.93 ± 18.49 ranged in 100.26 ~ 168.36 ㎣) were selected and used in the present study. CH were orally administered in a volume of 10 ㎖/kg (of body weight), once a day for 35 days from 21 days after tumor cell implantation, as equivalence 400, 200 and 100 mg/kg. Tamoxifen was also orally administered in a volume of 10 ㎖/kg as equivalence 20 mg/kg, in the present experiment. In intact and TB controls, instead of test solutions, only distilled water was also orally administered in a volume of 10 ㎖/kg. Groups (Six groups, eight mice in each group) 1. Intact control : Vehicle (distilled water 10 ㎖/kg) administered intact athymic nude mice 2. Tumor bearing (TB) control : Vehicle administered MCF-7 cell xenograft athymic nude mice 3. T20 : Tamoxifen 20 mg/kg administered MCF-7 cell xenograft athymic nude mice 4. CH400 : CH 400 mg/kg administered MCF-7 cell xenograft athymic nude mice 5. CH200 : CH 200 mg/kg administered MCF-7 cell xenograft athymic nude mice 6. CH100 : CH 100 mg/kg administered MCF-7 cell xenograft athymic nude mice RESULTS : As results of tumor cell xenograft in athymic nude mice, great decreases of spleen and submandibular lymph node weights, splenic TNF-α, IL-1β and IL-10 contents, serum IFN-γ splenic and peritoneal NK cell activities were observed in histopathological atrophic changes of submandibular amd spleen lymph nodes. Furthemore, decreases on the body weight and gains were also observed in TB control with increases of serum IL-6 levels, decreases of periovarian fat pad weights, atrophic changes of white adipose tissues, suggesting MCF-7 tumor cell inoculation related immunosuppress and cachexia. However, additional oral administration of all three different dosages of CH 400, 200 and 100 mg/kg effectively inhibited MCF-7 cell xenograft tumor mass growth, the volumes and weights at the day of terminal sacrifice, dose-dependently, more favorable or as comparable to those of tamoxifen 20 mg/kg in CH 400 mg/kg, at least in a condition of the current experiment. In addition, CH 400, 200 and 100 mg/kg administered mice also showed favorable and dose-dependent inhibitory activities on cancer-related immunosuppress and cachexia in MCF-7 xenograft mice, as compared with those of TB control mice. Marked decreases of tumor volumes and weights were also demonstrated in tamoxifen 20 mg/kg treated mice with decreases of tumor cell volumes, and also marked increases of tumor mass cleaved caspase-3 and PARP immunoreactive tumor cells and decreases of COX-2 immunoreactivities were observed in tamoxifen 20 mg/kg treated mice as compared to those of TB control mice. However, tamoxifen 20 mg/kg treatment deteriorated the cancer cachexia (more decreased in the body weight gains, periovarian fat depositions and elevation of serum IL-6 levels) and immunosuppress (more decrease of lymphatic organ weights, serum IFN-γ levels, splenic TNF-α NK cell activities, IL-10 and IL-1β contents, histopathological atrophic changes of lymphatic organs) as compared with TB control mice, without meaningful changes on tumor mass iNOS and TNF-α immunoreactivities, at least in a condition of this experiment. CONCLUSION : Based on the current results of MCF-7, one of the most frequently used human breast cancer derived adenocarcinoma cell line xenograft athymic nude mice, it is considered that oral administration of CH at dose levels of 400, 200 and 100 mg/kg, markedly and dose-dependently showed favorable anti-tumor activities, through their potent immunomodulatory activities, more favorable or as comparable to those of tamoxifen 20 mg/kg in CH 400 mg/kg, and they also effectively controlled related cancer cachexia, which were deteriorated by tamoxifen 20 mg/kg oral administration, at least in a condition of the current study. It, therefore, expected that appropriate oral administration of CH, will be provide effective and novel anti-tumor alternative therapeutic regimes, including cancer cachexia control without serious side effects, through more detail mechanism and clinical trials with screening of the biological active compounds, in future.

Combined Treatment of Sodium Salicylate and Genistein Induces Incomplete Apoptosis and Necrosis in MCF-7 Multicellular Tumor Spheroids

Su Yeon Lee(이수연),Cho Hee Kim(김초희),Hyun Min Jeon(전현민),Min Kyung Ju(주민경),Min Young Kim(김민영),Eui-kyong Jeong(정의경),Hye Gyeong Park(박혜경),Ho Sung Kang(강호성) 한국생명과학회 2012 생명과학회지 Vol.22 No.9

아스피린과 아스피린의 deacetylated form인 sodium salicylate (NaSal)은 대장암, 폐암 및 유방암을 비롯한 다양한 암의 항암제 활성을 나타내는 것으로 잘 알려져 있다. A549 폐암 세포주에 저농도의 NaSal과 genistein을 함께 복합 처리시 상승작용에 의해 세포사멸을 증가시켜서 NaSal에 의한 암억제 효과를 증대시킴을 이미 밝힌 바 있다. 본 연구에서는 A549가 아닌 다른 암세포주와 in vitro solid tumor model인 multicellular spheroids (MTS)을 이용하여 NaSal과 genistein 복합처리 효과를 조사하였다. NaSal/genistein 복합 처리시 A549 세포주와 마찬가지로 HCT116 세포주에서도 세포사멸이 유도되었지만, MCF-7 세포주에서는 유도되지 않았다. 흥미롭게도, MCF-7 세포주는 MTS로 배양되는 동안 NaSal/genistein 복합 처리에 의해 세포 죽음을 나타내었다. 세포 죽음의 형태는 MCF-7 MTS의 발달 단계에 따라 세포사멸 또는 세포괴사로 나타났다. MCF-7 MTS에서의 세포사멸은 불완전한 양상을 보였다. 즉 염색체가 응축되고 쪼개지지만, 핵막은 여전히 관찰되었다. 이상의 연구 결과 NaSal/genistein 복합처리는 MCF-7 MTS 배양 system에서 불완전한 세포사멸과 세포괴사를 일으킴을 알 수 있었다. Aspirin and its deacetylated form, sodium salicylate (NaSal), have been shown to exert chemopreventive activities against many human cancers including those of the colon, lung, and breast. Previously, we showed that combined treatment of NaSal and genistein synergistically induced apoptosis in A549 lung cancer cells, indicating that these two natural chemicals could be used in combination for cancer therapy. In this study, we examined effects of NaSal/genistein combined treatment on other cancer cells and in three-dimensional multicellular tumor spheroid (MTS) and in an in vitro solid tumor model. We found that the combined treatment induces apoptosis in the HCT116 cells and the A549 cells, but not in the MCF-7 cells. Interestingly, the MCF-7 cells responded to the NaSal/genistein combined treatment by undergoing cell death when they were cultivated as MTS. The combined treatment induced apoptosis at an earlier stage in the MCF-7 MTS culture. However, when the MCF-7 MTS was cultivated for a longer period, it induced necrosis rather than apoptosis. We further found that the apoptotic pattern observed in MCF-7 MTS was incomplete: the chromatins were condensed and fragmented, but the nuclear membrane was still intact. Taken together, these results demonstrate that the NaSal/genistein combined treatment induces incomplete apoptosis and necrosis in the MCF-7 MTS culture system.

담즙산 합성유도체(HS-1200)가 인체 유방암 세포주(MCF-7)에서 유도하는 방사선 감작 효과

이형식(Hyung Sik Lee),최영민(Young Min Choi),권혁찬(Hyuk Chan Kwon),송연숙(Yeon Suk Song) 대한방사선종양학회 2004 Radiation Oncology Journal Vol.22 No.2

목 적: 인체 유방암 세포주인 MCF-7에 새로운 CDCA 합성유도체인 HS-1200을 방사선과 함께 처치하여 아포토시 스 유도 활성 및 방사선 감작 효과를 관찰하고자 하였다. 대상 및 방법: MCF-7 세포에 2∼8 Gy의 X-ray와 16μM 농도의 HS-1200을 처리한 세포들의 세포 생존 곡선을 clonogenic assay를 통하여 구하였다. 아포토시스 유도 확인은 8 Gy의 X-ray와 40μM 농도의 HS-1200을 전 처치하여 구한 agarose gel 전기영동 및 Hoechst staining을 이용하였다. 면역형광법을 이용한 cytochrome c, Bax 및 AIF들의 관찰과 미토콘드리아 막전위 측정을 시행하였다. Western blotting을 통한 PARP (poly (ADP-ribose) polymerase) cleavage, Bax, Bcl-2, Bak 및 AIF 들의 발현을 관찰하였다. 결 과: 2∼8 Gy의 X-ray를 조사한 군(R)과 HS-1200 처리 후 2∼8 Gy의 X-ray를 조사 한 군(HR)의 세포 생존 곡선을 비교하니 HR 군에서 세포 감작 효과를 관찰할 수 있었다. DNA ladder는 R군에서는 72 시간째 관찰되는 반면에 HR 군에서는 24시간째 관찰되어 DNA 분절이 빠르게 진행됨을 알 수 있었고, PARP cleavage의 관찰에서도 R 군에 비해 24시간 빠르게 진행되었다. 면역 형광법을 이용한 실험에서도 HR 군이 R 군에 비하여 미토콘드리아 막전위(ΔΨm)의 급격한 감소, cytochrome의 다량 방출, Bax의 증가된 점상 변화 등이 관찰되었고, AIF의 변화는 뚜렷하 지 않았다. Western blotting을 이용한 Bax, Bcl-2, Bak 및 AIF들의 발현을 관찰하였을 때 Bax 만 HR 군에서 시간대별로 증가되는 추세를 보인 반면 Bcl-2, Bak 및 AIF들의 발현은 특이한 차이를 발견할 수 없었다. 결 론: 인체 유방암 세포주(MCF-7)에서 새로운 담즙산 합성 유도체인 HS-1200은 방사선 조사에 의한 아포토시스의 유도를 감작시키는 사실을 관찰하였다. 아포토시스 유도감작 증가는 Bax/Bcl-2 분율의 상대적 증가로 기인한다고 생각한다. 상기 결과를 토대로 HS-1200의 항암 치료제로서의 역할에 관한 기초 자료로서의 유용성을 제시할 수 있었다. 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 Methods: Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37oC 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 16μM 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 40μM 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 40μM of HS-1200 combined with 8 Gy irradiation showed severalchanges associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200combined 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 (ΔΨm) and increased cytochromec 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.

Salubrinal-Mediated Upregulation of eIF2α Phosphorylation Increases Doxorubicin Sensitivity in MCF-7/ADR Cells

Jeon, Yong-Joon,Kim, Jin Hyun,Shin, Jong-Il,Jeong, Mini,Cho, Jaewook,Lee, Kyungho Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.2

Eukaryotic translation initiation factor 2 alpha ($eIF2{\alpha}$), which is a component of the eukaryotic translation initiation complex, functions in cell death and survival under various stress conditions. In this study, we investigated the roles of $eIF2{\alpha}$ phosphorylation in cell death using the breast cancer cell lines MCF-7 and MCF-7/ADR. MCF-7/ADR cells are MCF-7-driven cells that have acquired resistance to doxorubicin (ADR). Treatment of doxorubicin reduced the viability and induced apoptosis in both cell lines, although susceptibility to the drug was very different. Treatment with doxorubicin induced phosphorylation of $eIF2{\alpha}$ in MCF-7 cells but not in MCF-7/ADR cells. Basal expression levels of Growth Arrest and DNA Damage 34 (GADD34), a regulator of $eIF2{\alpha}$, were higher in MCF-7/ADR cells compared to MCF-7 cells. Indeed, treatment with salubrinal, an inhibitor of GADD34, resulted in the upregulation of $eIF2{\alpha}$ phosphorylation and enhanced doxorubicin-mediated apoptosis in MCF-7/ADR cells. However, MCF-7 cells did not show such synergic effects. These results suggest that dephosphorylation of $eIF2{\alpha}$ by GADD34 plays an important role in doxorubicin resistance in MCF-7/ADR cells.

Salubrinal-Mediated Upregulation of eIF2 Phosphorylation Increases Doxorubicin Sensitivity in MCF-7/ADR Cells

이경호,Yong-Joon Jeon,Jin-Hyun Kim,Jong-Il Shin,정민이,조재욱 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.2

Eukaryotic translation initiation factor 2 alpha (eIF2), which is a component of the eukaryotic translation initia-tion complex, functions in cell death and survival under various stress conditions. In this study, we investigated the roles of eIF2 phosphorylation in cell death using the breast cancer cell lines MCF-7 and MCF-7/ADR. MCF-7/ADR cells are MCF-7-driven cells that have acquired resistance to doxorubicin (ADR). Treatment of doxorubicin reduced the viability and induced apoptosis in both cell lines, although susceptibility to the drug was very different. Treatment with doxorubicin induced phosphorylation of eIF2 in MCF-7 cells but not in MCF-7/ADR cells. Basal expression levels of Growth Arrest and DNA Damage 34 (GADD34), a regulator of eIF2, were higher in MCF-7/ADR cells compared to MCF-7 cells. Indeed, treatment with salubrinal, an inhibitor of GADD34, resulted in the upregulation of eIF2 phosphorylation and enhanced doxorubicin-mediated apoptosis in MCF-7/ADR cells. However, MCF-7 cells did not show such synergic effects. These results suggest that dephosphorylation of eIF2 by GADD34 plays an important role in doxorubicin resistance in MCF-7/ADR cells.

항암제 내성 유방암 MCF7/adR 세포주에 대한 보정방암탕과 홍삼산성다당체의 세포고사 유도효과

안귀인,박철환,이은옥,이효정,이재호,김관현,이연희,장유성,김상태,김성훈,Ahn, Gyu-In,Park, Cheol-Hwan,Lee, Eun-Ok,Lee, Hyo-Jung,Lee, Jae-Ho,Kim, Kwan-Hyun,Rhee, Yun-Hee,Jang, Yu-Sung,Kim, Sang-Tae,Kim, Sung-Hoon 대한약학회 2006 약학회지 Vol.50 No.4

This study was undertaken to determine whether the 9 herbal complex induces apoptosis in human breast cancer MCF-7 cells and adriamycin-resistant MCF7/adR cells. Ethanol extracts of Bojungbangamtang (BBTE) and acidic polysaccharide of Red Ginseng (GIN) induced cell death in both MCF-7 and MCF7/adR cells. Ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng also induced $G_2/M$ cell cycle arrest and increased TUNEL positive cells in MCF7/adR cells. In addition, flow cytometric analysis revealed the decreased expression of P-glycoprotein (P-gp) in ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng treated MCF7/adR cells. Similarly, decreased protein levels of P-glycoprotein and multidrug resistance associated proteins-1 were also determined by immunocytometry in ethanol extracts of Bojungbangamtang treated MCF7/adR cells. Taken together these data indicate that ethanol extracts of Bojungbangamtang and acidic polysaccharide of Red Ginseng inhibit the function of ABC transporters such as multi drug resistance associated proteins (MRPs) and P-glycoprotein as well as induce apoptosis in MCF7/adR cells. Thus, these data suggest that ethanol extracts of Bojungbangamtang and polysaccharide of Red Ginseng can be candidates for the treatment of multidrug-resistant MCF7/adR cells.

Exogenous and Endogeneous Disialosyl Ganglioside GD1b Induces Apoptosis of MCF-7 Human Breast Cancer Cells

Ha, Sun-Hyung,Lee, Ji-Min,Kwon, Kyung-Min,Kwak, Choong-Hwan,Abekura, Fukushi,Park, Jun-Young,Cho, Seung-Hak,Lee, Kichoon,Chang, Young-Chae,Lee, Young-Choon,Choi, Hee-Jung,Chung, Tae-Wook,Ha, Ki-Tae,Ch MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.5

<P>Gangliosides have been known to play a role in the regulation of apoptosis in cancer cells. This study has employed disialyl-ganglioside GD1b to apoptosis in human breast cancer MCF-7 cells using exogenous treatment of the cells with GD1b and endogenous expression of GD1b in MCF-7 cells. First, apoptosis in MCF-7 cells was observed after treatment of GD1b. Treatment of MCF-7 cells with GD1b reduced cell growth rates in a dose and time dependent manner during GD1b treatment, as determined by XTT assay. Among the various gangliosides, GD1b specifically induced apoptosis of the MCF-7 cells. Flow cytometry and immunofluorescence assays showed that GD1b specifically induces apoptosis in the MCF-7 cells with Annexin V binding for apoptotic actions in early stage and propidium iodide (PI) staining the nucleus of the MCF-7 cells. Treatment of MCF-7 cells with GD1b activated apoptotic molecules such as processed forms of caspase-8, -7 and PARP (Poly(ADP-ribose) polymerase), without any change in the expression of mitochondria-mediated apoptosis molecules such as Bax and Bcl-2. Second, to investigate the effect of endogenously produced GD1b on the regulation of cell function, UDP-gal: β1,3-galactosyltransferase-2 (GD1b synthase, Gal-T2) gene has been transfected into the MCF-7 cells. Using the GD1b synthase-transfectants, apoptosis-related signal proteins linked to phenotype changes were examined. Similar to the exogenous GD1b treatment, the cell growth of the GD1b synthase gene-transfectants was significantly suppressed compared with the vector-transfectant cell lines and transfection activated the apoptotic molecules such as processed forms of caspase-8, -7 and PARP, but not the levels of expression of Bax and Bcl-2. GD1b-induced apoptosis was blocked by caspase inhibitor, Z-VAD. Therefore, taken together, it was concluded that GD1b could play an important role in the regulation of breast cancer apoptosis.</P>

MCF-7 cell에서 all-trans retinoic acid에 의한 insulin-like growth factor-I와 insulin-like growth factor binding protein-3 분비조절에 있어서 PKC-δ의 역할

이선미,김상훈,최광수,강창원,Lee, Sun-Mi,Kim, Sang-Hoon,Choi, Kwang-Soo,Kang, Chang-Won 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.2

All-trans retinoic acid (AtRA) induces growth inhibition and apoptosis in a variety of tumer cells, including MCF-7 cells. Insulin-like growth factors (IGFs) system has been reported to be associated with the development of cancer. Although MCF-7 cell with AtRA is to be the major stimulus for the cell growth and apoptosis, the mechanism of insulin-like growth factor-I (IGF-I)/insulin-like growth factor binding protein-3 (IGFBP-3) system remains to be elucidated. Thus, this study was conducted to the effect of AtRA on the gene expression and level of IGF-I and IGFBP-3. In addition, we investigated the involvement of PKC-${\delta}$ on the IGF-I and IGFBP-3 secretion in MCF-7 cell. AtRA(${\geq}10^{-7}M$) decreased the IGF-1 secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cells. Especially, the treatment of AtRA at 72 hours caused a significant reduction in the IGF-I secretion and mRNA expressions but increment in IGFBP-3 secretion and mRNA expressions (p < 0.05). $10^{-7}M$ AtRA activated PKC-${\delta}$ that is one among PKC-$\iota$, ${\alpha}$, ${\lambda}$ and ${\delta}$ in MCF-7 cell. Rotllerin, a PKC-${\delta}$ inhibitor, blocked AtRA-induced inhibition of the IGF-I and mRNA expressions, and increase of lGFBP-3 and mRNA expressions in MCF-7 cell. Together, AtRA inhibited the IGF-I secretion and mRNA expressions, but increased IGFBP-3 secretion and mRNA expressions in MCF-7 cell. Furthermore, AtRA-induced alteration of IGF-I, IGFBP-3 secretion, and the gene expressions were mediated via PKC-${\delta}$ activity.