Hep3B 간암세포에서 셀레늄과 커큐민의 병행처리에 의한 Akt와 mTOR 조절

이솔화,박송이,김인섭,박옥진,김영민 대한암예방학회 2010 Journal of cancer prevention Vol.15 No.4

Phytochemical in fruits and vegetables induced apoptosis with inhibition of cancer cell growth and metastasis. Selenium and curcumin, a kind of phytochemical, have anti-cancer and anti-oxidant effect in cancer cells. Selenium inhibits metastasis through the induction of cell cycle arrest. Curcumin, a major activate component turmeric, is known inflammatory, anti-microbial and anti-cancer effect. The ser/thr kinase Akt is activated by growth factor, and it controls cell survival and cancer cell invasion and metastasis. Mammalian target of rapamycin (mTOR) is modulated by AMP-activated protein kinase (AMPK). It regulates cell survival and growth. In this study, we have investigated the synergistic effects of selenium and curcumin on apoptosis and Akt and mTOR activation in Hep3B hepato-carcinoma cells. Treatment with selenium and curcumin or co-treatment with them inhibited proliferation and survival of Hep3B cells. They induced also apoptotic cell death of cancer cells. We suggest that Akt/mTOR signal pathway is inhibited more effective by combined treatment of selenium and curcumin in Hep3B hepatocarcinom cells, so proliferation of Hep3B cells can be suppressed. (Cancer Prev Res 15, 285-290, 2010)

MCF-7 유방암세포에서 레스베라트롤에 의한 TNF-α, Akt 및 COX-2 신호분자의 조절

이솔화,서동익,김혜선,박옥진,김영민,박송이 대한암예방학회 2012 Journal of cancer prevention Vol.17 No.2

Resveratrol, a naturally occurring polyphenol, has broad-spectrum beneficial effects including anti- proliferation, anti-inflammatory and anti-cancer activities. TNF-α is highly expressed in cancer cells. Akt is also highly expressed in breast cancer and regulates cell proliferation and survival. COX-2 is increased by prostaglandins, which includes cell proliferation and angiogenesis. In this study, we explored the anti-proliferative effects of resveratrol via inhibition of TNF-α, Akt and COX-2 in MCF-7 breast cancer cells. In MTT assay, resveratrol inhibits proliferation of MCF-7 breast cancer cells in does-dependent and time-dependent manner. Resveratrol also induced apoptotic cell death of cancer cells. Resveratrol decreased anti-apoptotic protein (Bcl-2) and increased pro-apoptotic protein (Bax, caspase-3 and PARP). Treatment with resveratrol decreased TNF-α in cytosol and nucleus, reduced Akt and COX-2 expression in the nucleus on MCF-7 breast cancer cell. These results suggest that resveratrol induced apoptosis through inhibition of TNF-α, Akt and COX-2 in MCF-7 breast cancer cells.

레스베라트롤의 HT-29 대장암 세포증식 및 이동성 억제효과

이솔화(Sol Hwa Lee),박송이(Song Yi Park),김인섭(In-Seop Kim),박옥진(Ock Jin Park),김영민(Young Min Kim) 한국생물공학회 2012 KSBB Journal Vol.27 No.5

Resveratrol, natural polyphenol in grapes and red wine, is known to have the anti-proliferatory and anti-angiogenic effects in various cancer cells. In this study, we have investigated the effects of resveratrol in HT-29 colon cancer cells. Treatment of resveratrol in different concentrations and time inhibited proliferation of HT-29 colon cancer cells. We explored the effects of resveratrol on HT-29 colon cancer cell motility using a wound healing assay. In the absence of the resveratrol, the HT-29 cells are migrated along the edges of the wound and showed a large-scale migration, whereas dose- and time-dependent inhibition of cell flattening and spreading was observed in the presence of resveratrol. Resveratrol inhibited MMP-9 in a dose- and time-dependent on HT-29 colon cancer cells by Western blotting. In addition, resveratrol increased AMPK activity and decreased COX-2, VASP and VEGF expression. Treatment of compound C inhibited AMPK activity, however, the expression of VASP and COX-2 increased thus, COX-2 and VASP are modulated by AMPK. However treatment of celecoxib could not control AMPK activity but decreased VEGF expression. We suggest that resveratrol inhibits cell proliferation and migration through activation of AMPK and decreased COX-2, VASP and VEGF expression in HT-29 colon cancer cells.

Apoptotic Effects of Resveratrol via mTOR and COX-2 Signal Pathways in MCF-7 Breast Cancer Cells

Sol Hwa Lee(이솔화),Hye yeon Lee(이혜연),Song Yi Park(박송이),Ock Jin Park(박옥진),Young Min Kim(김영민) 한국생명과학회 2011 생명과학회지 Vol.21 No.9

식물에서 추출한 파이토케미컬은 암세포의 여러 신호전달 기작에 관여함으로써 apoptosis를 유도한다. 본 연구에서는 파이토케미컬의 한 종류인 레스베라트롤을 MCF-7 세포에 처리함으로써 암세포의 증식 억제와 apoptosis 유도 효과를 알아보았고, 이러한 효과가 암세포의 성장과 증식에 관여하는 단백질인 mTOR와 COX-2의 발현 양상에 어떠한 영향을 미치는지 알아보고자 하였다. 그 결과 MCF-7 세포에 레스베라트롤을 처리했을 때 농도가 증가함에 따라 암세포의 생존률이 감소하였고, Hoechst 33342를 이용한 chromatin 염색과 Annexin V-propodium iodide staning을 통하여 암세포의 세포증식 효과 apoptosis에 의해 유도된 것임을 알 수 있었다. MCF-7 세포에 레스베라트롤을 처리했을 때 mTOR 및 COX-2의 발현 양상을 확인하기 위해 Western blotting을 실시한 결과, 레스베라트롤의 농도가 높아짐에 따라 mTOR 및 COX-2의 발현이 감소함을 확인 하였다. 이와 같은 결과는 MCF-7 유방암 세포에서 레스베라트롤에 의한 암세포의 증식 억제 및 apoptosis 유도가 mTOR 신호경로 저해를 통한 COX-2의 발현을 감소시킴으로써 나타나는 것으로 보인다. Resveratrol, a kind of phytochemical, is presented in grape skins. Resveratorl exerts antiproliferative, anti-cancer and pro-apoptotic activities in cancer cells. Mammalian target of rapamycin (mTOR) is a critical regulator of cellular growth and proliferation, and it is known to be a strategic target for anti-cancer therapeutic uses. mTOR is a major downstream of the PI3K/Akt pathway, which is activated in various cancer cells. It also plays an important role in the survival, proliferation and angiogenesis of cells. Cyclooxygenase-2 (COX-2) is an important protein that mediates inflammatory processes. It plays an important role in various tumors by affecting cell proliferation, mitosis, apoptosis and angiogenesis. In this study, we have investigated the effects of resveratrol on apoptosis through mTOR and COX-2 expression in MCF-7 breast cancer cells. The treatment of resveratrol with different concentrations inhibited proliferation of MCF-7. The data showed that resveratrol induced apoptotic cell death of cancer cells and decreased mTOR and COX-2 expression. These results suggest that resveratrol induces apoptosis of MCF-7 breast cancer cells by inhibiting mTOR and COX-2 expression.

MCF-7 유방암 세포에서 Lactic Acid Bacteria를 처리하였을 때 AMP-Activated Protein Kinase 활성과 Apoptosis 유도에 관한 연구

김윤이,이솔화,박옥진,김영민 대한암예방학회 2010 Journal of cancer prevention Vol.15 No.3

Lactic Acid Bacteria (LAB) and cultured dairy products exert effect of anti-cancer that inhibits the growth of many types of cancer, including breast cancer. Also, they exert effect of anti-metastasis that inhibits proliferation of cancer cells. AMP-activated protein kinase (AMPK) is activated by ATP depletion status such as heat shock, exercise, hypoxia, reactive oxygen species (ROS). Recent studies indicate that AMPK activation strongly suppresses cell proliferation and induces apoptosis in cancer cells. Hence,AMPK plays a critical role as a key signal in prevention of cancer. The AMPK activation is also known to be related with expression of cyclooxygenase-2 (COX-2) and mammalian target of rapamycin (mTOR). In this study, the LAB in MCF-7 breast cancer cell lines seemed to be closely related to an increase in AMPK activation. ROS generated by treatment of LAB, which induced apoptosis and inhibited proliferation of cancer cells. ROS is an upstream regulator of AMPK, and the activation of AMPK as the key element in the regulation of COX-2 and mTOR expression. Our results suggest that LAB is an effective way in controlling proliferation of breast cancer cells and apoptosis of them. Therefore, the effect of LAB will be a potent a chemotherapy in cancer.

MCF-7 유방암 세포에서 COX-2/VASP신호분자 조절을 통한 커큐민의 Apoptosis효과

이세희,김영민,이기호,박송이,이솔화,박옥진,구봉성 대한암예방학회 2012 Journal of cancer prevention Vol.17 No.1

Curcumin, a phytochemical acquired to yellow curry, has potent anti-inflammatory and anti-oxidant activities. Cyclooxygenase-2 (COX-2) is inducible enzymes, and regulates the level of prostaglandins during inflammation. A vasodilatator-stimulated phosphoprotein (VASP) is connected to actin, and is activated by phosphorylation. In this study, we investigated the effect of curcumin on apoptosis and COX-2/VASP signaling pathway. We observed that curcumin inhibited cell proliferation and caused apoptosis in MCF-7 breast cancer cells. Treatment of compound C, an AMPK inhibitor, resulted in increased cell proliferation of cancer cells and showed that COX-2 and VASP are regulated by AMPK. Also, the co-treatment with celecoxib (COX-2 inhibitor) and curcumin resulted in decreased cell proliferation in breast cancer cells. Inhibition of COX-2 was regulated by the expression of VASP, indicating that the COX-2/VASP signaling molecules played an important role for apoptosis in MCF-7 breast cancer cells. In conclusion, curcumin had anti-proliferative and apoptotic effects through the regulation of COX-2and VASP signal pathway.

HCT116 대장암 세포에서 Akt-p53 신호경로를 통한 커큐민과 EGCG의 apoptosis 효과

Song-Yi Park(박송이),Sol-Hwa Lee(이솔화),Ock-Jin Park(박옥진),Young-Min Kim(김영민) 한국생명과학회 2011 생명과학회지 Vol.21 No.1

식품에서 추출한 파이토케미컬은 여러 암종에서 암세포의 증식억제와 apoptosis를 유도한다. 최근에 이러한 파이토케미컬의 세포 내 신호전달 기작에 관한 관심이 높아지고 있으며, 본 연구에서는 파이토케미컬의 일종인 커큐민과 EGCG를 HCT116 대장암세포에 처리함으로써 암세포의 증식억제와 apoptosis 유도 효과를 알아보고, 암세포의 증식에 관여하는 Akt의 활성과 종양 억제유전자인 p53의 신호경로를 규명하고자 하였다. 그 결과, 커큐민과 EGCG를 처리했을 때 HCT116 세포의 증식이 억제되었고, 암세포에서 apoptosis 효과가 나타남을 확인하였다. 동일한 조건에서 Western blotting을 실시했을 때 Akt의 활성은 감소하였으며 p53의 발현은 증가하였다. 또한 Akt의 저해제인 LY294002를 처리했을 때 암세포의 증식이 더욱 강하게 억제되었으며, p53의 발현은 더욱 강하게 증가하는 것으로 나타났다. 따라서 HCT116 세포에서 커큐민과 EGCG 처리에 의한 암세포의 증식 억제 및 apoptosis는 p53의 발현이 증가함에 따라 유도되며, 이러한 p53의 발현 증가는 Akt 신호경로를 저해함으로써 일어난다는 것을 확인하였다. p53 is tumor suppressor gene that regulates apoptosis such as caspase-dependent and p21-mediated signaling pathways. PI3K/Akt is known to be over-activated in cancer cells. Akt activates many survival-related signals such as mTOR and COX-2. Inactivation of Akt would result in non-inhibition of p53 as well as induced apoptosis. In this study, we showed that curcumin and EGCG activate p53 via inhibition of the Akt signaling pathway. Treatments using curcumin and EGCG in different concentrations for 24 hr and 48 hr inhibited proliferation of HCT116 colon cancer cells and increased apoptotic cell death. Also, our data showed that curcumin and EGCG increased the p53 expression and decreased the p-Akt. Treatment of LY294002 (Akt inhibitor) resulted in decreased cell proliferation of cancer cells, while LY294002 treated with curcumin or EGCG showed a greater decrease of cell proliferation. In addition, inhibition of Akt induced p53 activation in HCT116 colon cancer cells. These results suggest that curcumin and EGCG induce apoptosis by inhibiting Akt and increase p53 in HCT116 colon cancer cells.

HT-29 대장암세포에서 Akt 활성 저해에 따른 셀레늄의 세포 증식억제 효과

Song Yi Park(박송이),In-Seop Kim(김인섭),Se Hee Lee(이세희),Sol Hwa Lee(이솔화),Da Woon Jung(정다운),Ock Jin Park(박옥진),Young Min Kim(김영민) 한국생명과학회 2012 생명과학회지 Vol.22 No.1

Akt는 세포의 증식과 분화에 관여하며 많은 암종에서 과발현되어 있다는 것이 보고되었다. 본 연구에서는 Akt의 조절을 통한 셀레늄의 HT-29 세포의 세포증식억제 시너지효과를 확인하였다. 셀레늄을 농도별과 시간별로 처리하였을 때 HT-29 세포의 증식이 억제되었고, apoptosis가 일어남을 확인하였다. 셀레늄을 농도별로 처리하여 Western blotting 및 immunofluorescence를 실시한 결과 Akt의 인산화가 저해되었고 COX-2의 발현도 저해되었다. 또한 Akt 저해제인 LY294002를 처리한 결과, HT-29 대장암세포의 증식이 억제되었으며, LY294002를 셀레늄과 병행처리하였을 때 셀레늄에 의한 세포증식억제 효과가 더 강하게 나타나는 것을 확인하였다. Akt siRNA에 의한 Akt의 불활성화는 non-transfected 세포에 비하여 HT-29 세포의 성장을 더 강하게 억제하였으며, Akt가 불활성화 되었을 때 COX-2의 발현 역시 non-transfected 세포에 비하여 감소된 것을 확인하였다. 따라서 HT-29 세포에서 셀레늄의 세포증식억제 효과는 Akt와 COX-2 신호분자의 조절을 통해 일어나며, Akt 의 저해는 셀레늄의 대장암세포증식 억제에 시너지 효과를 나타냄을 확인하였다. Akt is known to play an important role in cell proliferation and differentiation, and is also over-expressed in several types of cancer cells. In this study, we explored the anti-proliferative effects of selenium in HT-29 colon cancer cells, mediated through effects on Akt and COX-2. Selenium treatments at different concentrations and for different durations inhibited proliferation of HT-29 colon cancer cells and increased apoptotic cell death. Selenium treatment decreased Akt phosphorylation and COX-2 expression. Treatment with LY294002 (an Akt inhibitor) decreased proliferation of HT-29 cells, while a combined treatment with LY294002 and selenium resulted in even further decreases in cell proliferation. Inactivation of Akt by Akt siRNA treatment abolished these inhibitory effects on cell growth. COX-2 expression decreased in Akt transfected cells compared to non-transfected cells. These results suggest that selenium induced both anti-proliferative and apoptotic effects by inhibiting Akt phosphorylation and COX-2 expression. Selenium treatment also appeared to induce synergistic anti-proliferative effects by inhibition of Akt in HT-29 colon cancer cells.