• Molecules and Cells
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• 제42권9호
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• pp.628-636
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• 2019

Altered genetic features in cancer cells lead to a high rate of aerobic glycolysis and metabolic reprogramming that is essential for increased cancer cell viability and rapid proliferation. Pyruvate kinase muscle (PKM) is a rate-limiting enzyme in the final step of glycolysis. Herein, we report that PKM is a potential therapeutic target in triple-negative breast cancer (TNBC) cells. We found that PKM1 or PKM2 is highly expressed in TNBC tissues or cells. Knockdown of PKM significantly suppressed cell proliferation and migration, and strongly reduced S phase and induced G2 phase cell cycle arrest by reducing phosphorylation of the CDC2 protein in TNBC cells. Additionally, knockdown of PKM significantly suppressed $NF-{\kappa}B$ (nuclear factor kappa-light-chain-enhancer of activated B cells) activity by reducing the phosphorylation of p65 at serine 536, and also decreased the expression of $NF-{\kappa}B$ target genes. Taken together, PKM is a potential target that may have therapeutic implications for TNBC cells.

• International Journal of Oral Biology
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• 제37권4호
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• pp.189-195
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• 2012

Resistance to the induction of apoptosis is a possible mechanism by which tumor cells can survive anti-neoplastic treatments. Melanoma is notoriously resistant to anti-neoplastic therapy. Previous studies have demonstrated focal adhesion kinase (FAK) overexpression in melanoma cell lines. Given its probable role in mediating resistance to apoptosis, many researchers have sought to determine whether the downregulation of FAK in melanoma cells would confer a greater sensitivity to anti-neoplastic agents. Genistein is a known inhibitor of protein-tyrosine kinase (PTK), which may attenuate the growth of cancer cells by inhibiting the PTK-mediated signaling pathway. This present study was undertaken to investigate the effect of genistein on the expression of FAK and cell cycle related proteins in the G361 melanoma cell line. Genistein was found to have a preferential cytotoxic effect on G361 melanoma cells over HaCaT normal keratinocytes. Genistein decreased the expression of 125 kDa phosphotyrosine kinase and the FAK protein in particular. Genistein treatment did not affect the expression of p53 in G361 cells in which p21 is upregulated. The expression of cyclin B and cdc2 was downregulated by genistein treatment. Taken together, our data indicate that genistein induces the decreased proliferation of G361 melanoma cells via the inhibition of FAK expression and regulation of cell cycle genes. This suggests that the use of genistein may be a viable approach to future melanoma treatments.

• 한국식품영양과학회지
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• 제33권8호
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• pp.1237-1245
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• 2004

브라질 기원인 신령 버섯 (A. blazei murill)은 강력한 항암 및 면역강화 작용을 가진 것으로 알려져 있다. 본 연구에서는 신령버섯 수용성 추출물(water extracted A. blazei Murill, WEAB)이 A549 인체 폐암세포 증식에 미치는 영향을 조사하였으며, 증식억제와 연관된 기전 해석을 시도하였다. WEAB가 처리된 A549 세포는 처리 농도 의존적으로 생존율이 감소되었으며, WEAB 처리는 암세포의 다양한 형태적 변형을 유발하였다. Flow cytometry 분석 결과로서 WEAB 처리에 의한 A549 폐암세포의 증식억제는 세포주기 G2/M arrest 및 apoptosis 유발과 직접적으로 연관성이 있음을 알 수 있었다. WEAB가 처리된 암세포에서 전사 및 번역 수준에서 cyclin A 발현의 감소 및 Cdk inhibitor p21 발현의 증가 현상이 관찰되었으나, cyclin B1, Cdk2, Cdc2, Wee1, Cdc25c 및 p53 등의 발현에는 큰 변화가 관찰되지 못하였다. 또한 WEAB의 처리는 COX-2 선택적 발현 저하를 유발하였으나, telomere 조절 관련 유전자들의 발현에는 큰 영향을 주지 못하였다. 이상의 결과는 신령버섯 추출물이 강력한 항암 및 암 예방 효능의 잠재력을 가지고 있음을 의미하며, 이에 관한 지속적인 연구가 필요할 것으로 생각된다.

• Molecular & Cellular Toxicology
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• 제2권3호
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• pp.159-165
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• 2006

To determine the anticancer effect of D-amygdalin (D-mandelinitrole-${\beta}$-D-gentiobioside) in human chronic myeloid leukemia cells K562, we profiled the gene expression between amygdalin treatment and control groups. Through 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, the cytotoxicity of D-amygdalin was $57.79{\pm}1.83%$ at the concentration of 5 mg/mL for 24 h. We performed cDNA microarray analysis and compared the gene expression profiles between D-amygdalin (5 mg/mL, 24 h) treatment and control groups. Among the genes changed by D-amygdalin, we paid attention to cell cycle-related genes, and particularly cell cycle regulator genes; because arrest of cell cycle processing was ideal tactic in remedy for cancer. In our data, expressions of cyclin-dependent kinase inhibitor 1B (p27, Kip1) (CDKN1B), ataxia telangiectasia mutated (includes complementation groups A, C, and D) (ATM), cyclin-dependent kinase inhibitor 1C (p57, Kip2) (CDKN1C), and CHK1 checkpoint homolog (CHEK1, formally known as CHK1) were increased, while expressions of cyclin-dependent kinase 2 (CDK2), cell division cycle 25A (CDC25A), and cyclin E1 (CCNE1) were decreased. The pattern of these gene expressions were confirmed through RT-PCR. Our results showed that D-amygdalin might control cell cycle regulator genes and arrest S phase of cell cycle in K562 cells as the useful anticancer drug.

• Biomolecules & Therapeutics
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• 제27권2호
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• pp.216-221
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• 2019

The c-Met protein is a receptor tyrosine kinase involved in cell growth, proliferation, survival, and angiogenesis of several human tumors. Overexpression of c-Met has been found in gastric cancers and correlated with a poor prognosis. Indirubin is the active component of Danggui Longhui Wan, which is a traditional Chinese antileukemic recipe. In the present study, we tested the anti-cancer effects of an indirubin derivative, LDD-1937, on human gastric cancer cells SNU-638. When we performed the in vitro kinase assay against the c-Met activity, LDD-1937 inhibited the activity of c-Met. This result was confirmed by immunoblot and immunofluorescence of phosphorylated c-Met. Immunoblot analysis showed that LDD-1937 decreased the expression of the Erk1/2, STAT3, STAT5, and Akt, downstream proteins of c-Met. In addition, LDD-1937 reduced the cell viability and suppressed colony formation and migration of SNU-638 cells. Furthermore, LDD-1937 induced $G_2/M$ phase arrest in the SNU-638 cells by decreasing the expression levels of cyclin B1 and CDC2. Cleaved-PARP, an apoptosis-related protein, was up-regulated in cells treated with LDD-1937. Overall, this study suggests that LDD-1937 may be a novel small-molecule with therapeutic potential for selectively inhibiting c-Met and c-Met downstream pathways in human gastric cancers overexpressing c-Met.

• BMB Reports
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• 제44권8호
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• pp.529-534
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• 2011

Ribosomal protein S3 (rpS3) is a multifunctional protein involved in translation, DNA repair, and apoptosis. The relationship between rpS3 and cyclin-dependent kinases (Cdks) involved in cell cycle regulation is not yet known. Here, we show that rpS3 is phosphorylated by Cdk1 in G2/M phase. Co-immunoprecipitation and GST pull-down assays revealed that Cdk1 interacted with rpS3. An in vitro kinase assay showed that Cdk1 phosphorylated rpS3 protein. Phosphorylation of rpS3 increased in nocodazole-arrested mitotic cells; however, treatment with Cdk1 inhibitor or Cdk1 siRNA significantly attenuated this phosphorylation event. The phosphorylation of a mutant form of rpS3, T221A, was significantly reduced compared with wild-type rpS3. Decreased phosphorylation and nuclear accumulation of T221A was much more pronounced in G2/M phase. These results suggest that the phosphorylation of rpS3 by Cdk1 occurs at Thr221 during G2/M phase and, moreover, that this event is important for nuclear accumulation of rpS3.

• 생명과학회지
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• 제16권4호
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• pp.589-597
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• 2006

본 연구에서는 T24 인체방광암 및 U937 백혈병 세포의 증식에 미치는 genistein의 영향을 조사 하였다. Genistein이 처리된 T24 및 U937 세포는 처리 농도 의존적으로 세포의 증식이 현저히 감소되었으며 심한 형태적 변형이 동반되었으나, U937 세포에서 보다 높은 감수성을 보였다. 이러한 T24 및 U937 세포의 증식억제 및 형태 변형은 G2/M기의 세포주기 억제 및 apoptosis 유발과 연관성이 있음을 flow cytometry를 이용한 세포주기의 분석을 통하여 확인하였다. T24 세포에서 genistein에 의한 G2/M arrest는 cyclin A, cyclin B1 및 Cdc25C 등의 단백질 발현 감소와 연관성이 있었으나, 종양억제 유전자 p53 및 Cdk inhibitor p21의 발현에는 큰 변화가 없었다. U937 세포에서 genistein에 의한 G2/M arrest는 cyclin B1 및 p53 비의존적인 p21의 발현 증가와 연관성이 있었다. 이상의 결과들은 현재까지 거의 연구가 진행된 바 없는 인체방광암 및 백혈병 세포에서 genistein의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 genistein을 포함한 그와 유사한 항암제 후보물질들의 연구에 있어서 기초 자료로서 사용될 수 있을 것으로 생각된다.

• 생명과학회지
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• 제15권3호
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• pp.397-405
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• 2005

브로콜리와 같은 십자화과 식물에서 glucoraphanin의 가수분해를 통해 생성되는 isothiocyanate의 일종인 sulforaphane은 강력한 항암효과를 가지며, 역학적 조사를 포함한 다양한 선행 연구에서 androgen 비 의존적으로 성장하는 전립선 암세포의 증식을 억제하는데 효과가 있었다. 최근 연구 결과에 따르면 sulforaphane은 다양한 인체암세포의 증식을 억제하고 apoptosis를 유발할 수 있는 것으로 알려지고 있으나, 정확한 분자생물학적 기전은 밝혀져 있지 않은 상태이다. 본 연구에서는 sulforaphane의 항암작용 기전을 조사하기 위하여 HeLa 인체자궁경부암세포의 증식에 미치는 sulforaphane의 영향을 조사하였다. Sulforaphane의 처리에 의한 HeLa 세포의 증식억제 및 형태적 변형은 세포주기 C2/M arrest 및 apoptosis 유발과 밀접한 관련이 있음을 알 수 있었다. RT-PCR 및 Western blot 분석 결과, sulforaphane 처리에 의하여 cyclin A 및 cyclin-dependent kinase (Cdk)4 단백질의 발현이 선택적으로 저하되었으며, Cdc2, Cdk inhibitor인 p16 및 p21의 발현은 증가되었다 그러나 sulforaphane은 cyclooxygenases의 발현이나 telomere 조절에 중요한 역할을 하는 인자들의 발현에는 큰 영향을 주지 못하였다. Sulforaphane의 항암 기전을 규명하기 위해서는 더 많은 연구가 부가적으로 필요하겠지만, 본 연구의 결과들에 의하면 sulforaphane은 강력한 인체암세포의 증식 억제 및 항암작용이 있을 것을 시사하여 준다고 할 수 있다.

• 생명과학회지
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• 제25권10호
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• pp.1184-1195
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• 2015

새로운 실험 동물로 대두되고 있는 제브라피쉬는 척추동물 생식생물학 연구에서도 중요한 역할을 한다. 제브라피쉬의 난자 성숙은 maturation inducing hormone (MIH, 17α,20β-Dihydroxy-4-pregnen-3-one)에 의해 촉발된다. 대부분의 동물의 난자성숙에는 cdc2 kinase와 cyclinB 단백질 복합체인 MPF의 활성화가 필요하다. 발톱개구리와 생쥐에서는 MPF 활성이 두 가지 기작에 의해 조절되는데, 하나는 cyclinB 결합이고 또 다른 하나는 Wee1과 Cdc25에 의한 T14/Y15 잔기의 억제성인산화와 탈인산화이다. 발톱개구리나 생쥐와 달리 제브라피쉬를 포함한 대부분의 진골어류(teleost)는 GV 난자에 pre-MPF complex가 존재하지 않으므로 MPF 활성화는 전적으로 cyclinB 단백질의 de novo synthesis에 의존한다. 다른 종과 마찬가지로 제브라피쉬의 모계유래 mRNA도 CPEB, Dazl, Pum1/Pum2, insulin-like growth factor2 mRNA-binding protein 3 등 다양한 RNA binding protein (RBP)의 결합에 의해 번역이 조절된다. 그러나 제브라피쉬 난자에서 단백질 번역 조절에 관여하는 자세한 작용 기작은 확실하게 규명되지 않았다. 그러므로 제브라피쉬 난자의 성숙과정을 연구하는 것은 척추동물 난자 초기 성숙과정에서 단백질 번역 조절의 역할을 규명할 수 있는 새로운 정보를 제공할 것이다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.110-113
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• 2005

Under the condition of nutritional deprivation, actively growing cells prepare to enter $G_0$-like stationary phase. Protein modification by phosphorylation/dephosphorylation or ubiqutination contributes to transfer cells from active cell cycle to dormant stage. We show here that Psp1/Sds23, which functions in association with the 20S cyclosome/APC (1) and is essential for cell cycle progression in Schizosaccharomyces pombe (2), is phosphorylated by stress-activated MAP kinase Sty1 and protein kinase A, as well as Cdc2/cyclinB, upon entry into stationary phase. Three serines at the positions 18,333 and 391 are phosphorylated and overexpression of Psp1 mutated on these sites causes cell death in stationary phase. These modifications are required for the binding of Spufd2, a S.pombe homolog of multiubiquitin chain assembly factor E4 in ubiquitin fusion degradation pathway. Deletion of Spufd2 gene led to increase cell viability in stationary phase, indicating that S. pombe Ufd2 functions to inhibit cell growth at this stage to maintain cell viability. Moreover, Psp1 enhances the multiubiquitination function of Ufd2, suggesting that Psp1 phosphorylated by sty1 and PKA kinases is associated with the Ufd2-dependent protein degradation pathway, which is linked to stress tolerance, to maintain cell viability in the $G_0$-like stationary phase.