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Rakib, Md. Abdur,Lee, Won Sup,Kim, Gon Sup,Han, Jae Hee,Kim, Jeong Ok,Ha, Yeong Lae Hindawi Publishing Corporation 2013 Evidence-based Complementary and Alternative Medic Vol.2013 No.-
<P>The major conjugated linoleic acid (CLA) isomers, <I>c</I>9,<I>t</I>11-CLA and <I>t</I>10,<I>c</I>12-CLA, have anticancer effects; however, the exact mechanisms underlying these effects are unknown. Evidence suggests that reversal of reduced gap junctional intercellular communication (GJIC) in cancer cells inhibits cell growth and induces cell death. Hence, we determined that CLA isomers enhance GJIC in human MCF-7 breast cancer cells and investigated the underlying molecular mechanisms. The CLA isomers significantly enhanced GJIC of MCF-7 cells at 40 <I><I>μ</I></I>M concentration, whereas CLA inhibited cell growth and induced caspase-dependent apoptosis. CLA increased connexin43 (Cx43) expression both at the transcriptional and translational levels. CLA inhibited nuclear factor-<I><I>κ</I></I>B (NF-<I><I>κ</I></I>B) activity and enhanced reactive oxygen species (ROS) generation. No significant difference was observed in the efficacy of <I>c</I>9,<I>t</I>11-CLA and <I>t</I>10,<I>c</I>12-CLA. These results suggest that the anticancer effect of CLA is associated with upregulation of GJIC mediated by enhanced Cx43 expression through inactivation of NF-<I><I>κ</I></I>B and generation of ROS in MCF-7 cells.</P>
Anti-proliferative Effects of β-ionone on Human Lung Cancer A-549 Cells
Sun Min Lee(이선민),Young Sook Kim(김영숙),Wook Jin Jang(장욱진),Abdur Md. Rakib(압두르 라키브),Tae Woo Oh(오태우),Boh Hyun Kim(김보현),So Young Kim(김소영),Jeong Ok Kim(김정옥),Yeong Lae Ha(하영래) 한국생명과학회 2013 생명과학회지 Vol.23 No.11
β-Ionone의 인체 비소폐암세포 A-549 cells (human non-small lung cancer A-549 cell)에 대한 anti-proliferative effect에 관한 연구를 수행하였다. A-549 cell에 다양한 농도의 β-ionone (1, 5, 10, and 15 μM)을 2, 4, 6일 간 처리 하고, 배양 2일에 A-549 cell의 생육억제와 관련되는 biochemical marker를 측정하였다. β-Ionone은 A-549 cell 생육을 dose와 time 의존적으로 저해하였다. β-Ionone 배양 2일의 IC50은 5.0 μg/ml이었다. β-Ionone은 농도 의존적으로 apoptosis를 유도하였다. β-Ionone은 p53, p21 및 Bax protein 수준을 증가시켰으나, Bcl-2 protein 발현은 억제시켰다. β-ionone은 cytosol cytochrome c 함량을 증가시켰고, caspase-9과 caspase-3 효소 활성증가를 유도하였다. 또한, β-ionone은 cPLA₂와 COX-2 protein level을 감소시켰다. 이와 같은 결과는 β-ionone의 A-549 cell에 대한 생육억제효과는 Bax와 Bcl-2 gene 발현을 reciprocal regulation하여 유도한 apoptosis와 cPLA₂ 및 COX-2 protein 발현 억제에 기인함을 의미한다. The anti-proliferative activity of β-ionone was investigated on human non-small lung cancer A-549 cells (designated A-549 cells). A-549 cells were treated with various concentrations of β-ionone (1, 5, 10, and 15 μM) for two, four, and six days. Biochemical markers related to the growth inhibition of A-549 cells by β-ionone were measured at the second day of incubation. β-Ionone inhibited the growth of A-549 cells by dose-and time-dependent manners, resulting in an IC50 of 5.0 μg/ml at the second day of incubation. β-Ionone induced apoptosis by a dose-dependent manner. β-Ionone increased levels of p53, p21, and Bax proteins, but suppressed expression of the Bcl-2 protein. Similarly, β-ionone enhanced cytochrome c release from the mitochondria to the cytosol, and induced activation of caspase-9 and -3. Additionally, β-ion-one reduced cPLA₂ and COX-2 protein levels. These results suggest that the β-ionone inhibits the proliferation of A-549 cells through reciprocal regulation of Bax and Bcl-2 gene expression and suppression of cPLA₂ and COX-2 protein expressions.
신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리
김소영(So Young Kim),김영숙(Young Suk Kim),장정순(Joung Soon Jang),김보현(Boh Hyun Kim),라키브 압두르(Abdur Md. Rakib),김곤섭(Gon Sup Kim),김정옥(Jeong Ok Kim),하영래(Yeong Lae Ha) 한국생명과학회 2014 생명과학회지 Vol.24 No.12
신령버섯균사체(Agaricus blazei mycelia: ABM)를 대두박이 함유된 액체배지에 배양하고, 이것을 자가분해(53 ℃, pH 5.5, 120 rpm, 3 hr)하여 항암성이 강한 isoflavone-conjugated glycoprotein (Gluvone 이라 명명)을 분리하였다. Gluvone 은 지금까지 알려진 당단백질과는 달리 분자량이 작고(9,400 Da), isoflavone이 결합되어 있다는 점이 다르다, Gluvone 은 60% 탄수화물(glucose, fructose, ribose), 31% 단백질 및 2% isoflavone (daidzein, genistein)으로 구성되어 있었다. 이 Gluvone은 S-180 복수암세포, MCF-7 인체유선암세포에 대한 독성이 강하였고, S-180 세포로 유발한 mouse 복수암을 강하게 억제하였다. Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone- conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at 53℃ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with ¹H-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.