• Biomolecules & Therapeutics
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• 제27권1호
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• pp.41-47
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• 2019

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated $IC_{50}$ value of $3{\mu}M$ after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G_1$ phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the $PERK/elF2{\alpha}/CHOP$ apoptotic pathway, and mitochondrial $Ca^{2+}$ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER- and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.

• Natural Product Sciences
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• 제15권1호
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• pp.1-7
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• 2009

This study was performed to investigate the cell growth inhibitory effect of tissue cultured root of wild Panax ginseng C.A. Mayer (tcwPG). The human stomach carcinoma cell line, MKN 74, was incubated with 70% EtOH extract of tcwPG or Panax ginseng C.A. Mayer (PG) for 24 hrs. tcwPG inhibited cell growth at a concentration of $250{\mu}g/ml$. However, Panax ginseng extract did not inhibit cell growth at the same concentration. We also tested the ethyl acetate and $H_2O$ fractions of tcwPG. The inhibitory effect of the ethyl acetate fraction on cell proliferation in MKN 74 cells was more potent than that of the crude extract, and the inhibitory effect of the $H_2O$ fraction was less than that of the ethyl acetate fraction. When we separated tcwPG into polar and non-polar saponin fractions and then measured cell growth inhibition, the non-polar saponin in tcwPG exhibited cytotoxicity. To compare the effects of tcwPG on various cancer cell lines, we measured cytotoxicity in MKN 74 (stomach cancer cell line), SW 620 (colon cancer cell line) and PC 3 (prostate cancer cell line). All three cell lines showed cell growth inhibition, and the cell growth inhibitory effects were not quite different in the various cell lines. The non-polar saponins of tcwPG arrested PC 3 cells at G1-phase as did Panax ginseng.

• 생명과학회지
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• 제27권10호
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• pp.1215-1224
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• 2017

현재까지 다양한 암의 발병 원인이 밝혀졌는데, 그 중 하나로써 DNA에 돌연변이가 축적되어 유전체가 불안정 해짐에 따라 암이 발생될 수 있는 기작들이 주목받고 있다. 생물정보학과 유전체학의 발달에 따라 질병 연구에 있어서 보다 더 정확하고 신뢰성 있는 바이오마커를 찾는 것이 가능해졌다. 따라서, 생물정보학과 유전체학의 연구 기반을 바탕으로 한 암의 바이오마커는 암의 조기진단뿐만 아니라, 더 나아가 암 발생 예측과 암환자의 예후 진단에 적용될 수 있다. 최근 들어 인간 유전체에서 약 45%를 차지하는 이동성 유전인자(transposable elements, TEs)가 유전자의 발현 조절과 DNA의 돌연변이를 유도함으로써 다양한 질병에 영향을 미친다는 사실이 밝혀짐에 따라, 이러한 이동성 유전인자들이 암의 발생과 어떤 연관이 있는지에 대한 연구 또한 활발히 진행되고 있다. 따라서 우리는 이동성 유전인자가 대장암과 어떤 연관성이 있는지에 대해 조사를 하였으며, 이를 어떻게 바이오마커로 활용할 수 있는지 알아보았다. 우선, 이동성 유전인자 중 인간 유전체에 많이 존재하면서 유전체에 많은 영향을 미치는 LINE-1 (long interspersed nuclear element-1, L1)과 Alu, LTR (long terminal repeat) 위주로 확인하였다. 흥미롭게도, 대장암 세포에서 LINE-1의 저메틸화, APC 유전자 내에 LINE-1 삽입, Alu의 저메틸화와 과메틸화, LTR 삽입에 따른 isoform 발생 등이 특징적으로 나타나는 것을 알 수 있었다. 또한 원발암유전자에서의 L1 저메틸화가 대장암 전이의 바이오마커로 쓰일 수 있다는 것과 Alu에 의한 MLH1 돌연변이가 가족성 및 유전성 대장암에서 흔히 발견된다는 것을 알 수 있었다. 이 때 이동성 유전인자에 의하여 영향 받는 유전자들의 발현을 in silico 발현 분석을 통하여 분석하였으며, 조직별, 성별 특이적 발현 양상을 제시하였다. 이들을 토대로 대장암 바이오마커를 개발하여 유전성 대장암의 예측 및 대장암 진단 또는 대장암 예후 예측을 통한 개인 맞춤형 치료에 활용할 수 있을 것으로 예상된다.

• Biomolecules & Therapeutics
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• 제30권2호
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• pp.137-144
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• 2022

Radiation resistance represents an imperative obstacle in the treatment of patients with colorectal cancer, which remains difficult to overcome. Here, we explored the anti-proliferative and migration-inhibiting properties of the natural product shikonin on a radiation-resistant human colon carcinoma cell line (SNU-C5RR). Shikonin reduced the viability of these cells in a dose-dependent manner; 38 µM of shikonin was determined as the half-maximal inhibitory concentration. Shikonin induced apoptotic cell death, as demonstrated by increased apoptotic body formation and the number of TUNEL-positive cells. Moreover, shikonin enhanced mitochondrial membrane depolarization and Bax expression and also decreased Bcl-2 expression with translocation of cytochrome c from mitochondria into the cytosol. In addition, shikonin activated mitogen-activated protein kinases, and their specific inhibitors reduced the cytotoxic effects of shikonin. Additionally, shikonin decreased the migration of SNU-C5RR cells via the upregulation of E-cadherin and downregulation of N-cadherin. Taken together, these results suggest that shikonin induces mitochondria-mediated apoptosis and attenuates epithelial-mesenchymal transition in SNU-C5RR cells.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3113-3121
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• 2014

Colorectal cancer remains one of the most common types of cancer and a leading cause of cancer death worldwide. In this study, we aimed to investigate effects of DuP-697, an irreversible selective inhibitor of COX-2 on colorectal cancer cells alone and in combination with a promising new multi-targeted kinase inhibitor E7080. The HT29 colorectal cancer cell line was used. Real time cell analysis (xCELLigence system) was conducted to determine effects on colorectal cell proliferation, angiogenesis was assessed with a chorioallantoic membrane model and apoptosis was determined with annexin V staining. We found that DuP-697 alone exerted antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells. For the antiproliferative effect the half maximum inhibition concentration ($IC_{50}$) was $4.28{\times}10^{-8}mol/L$. Antiangiogenic scores were 1.2, 0.8 and 0.5 for 100, 10 and 1 nmol/L DuP-697 concentrations, respectively. We detected apoptosis in 52% of HT29 colorectal cancer cells after administration of 100 nmol/L DuP-697. Also in combination with the thyrosine kinase inhibitor E7080 strong antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells were observed. This study indicates that DuP-697 may be a promising agent in the treatment of colorectal cancer. Additionally the increased effects observed in the combination with thyrosine kinase inhibitor give the possibility to use lower doses of DuP-697 and E7080 which can avoid and/or minimize side effects.

• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4651-4657
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• 2014

The development of chemopreventive approaches using a concoction of phytochemicals is potentially viable for combating many types of cancer including colon carcinogenesis. This study evaluated the anti-proliferative effects of ginger and Gelam honey and its efficacy in enhancing the anti-cancer effects of 5-FU (5-fluorouracil) against a colorectal cancer cell line, HCT 116. Cell viability was measured via MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphenyl)-2H-tetrazolium) assay showing ginger inhibiting the growth of HCT 116 cells more potently ($IC_{50}$ of 3mg/mL) in comparison to Gelam honey ($IC_{50}$ of 75mg/mL). Combined treatment of the two compounds (3mg/mL ginger+75mg/mL Gelam honey) synergistically lowered the $IC_{50}$ of Gelam honey to 22mg/mL. Combination with 35 mg/mL Gelam honey markedly enhanced 5-FU inhibiting effects on the growth of HCT 116 cells. Subsequent analysis on the induction of cellular apoptosis suggested that individual treatment of ginger and Gelam honey produced higher apoptosis than 5-FU alone. In addition, treatment with the combination of two natural compounds increased the apoptotic rate of HCT 116 cells dose-dependently while treatment of either ginger or Gelam honey combined with 5-FU only showed modest changes. Combination index analysis showed the combination effect of both natural compounds to be synergistic in their inhibitory action against HCT 116 colon cancer cells (CI 0.96 < 1). In conclusion, combined treatment of Gelam honey and ginger extract could potentially enhance the chemotherapeutic effect of 5-FU against colorectal cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1739-1744
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• 2016

We report herein an in vitro anticancer evaluation of a series of seven curcumin analogues (3a-g). The National Cancer Institute (NCI US) Protocol was followed and all the compounds were evaluated for their anticancer activity on nine different panels (leukemia, non small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer) represented by 60 NCI human cancer cell lines. All the compounds showed significant anticancer activity in one dose assay (drug concentration $10{\mu}M$) and hence were evaluated further in five dose assays (0.01, 0.1, 1, 10 and $100{\mu}M$) and three dose related parameters $GI_{50}$, TGI and $LC_{50}$ were calculated for each (3a-g) in micro molar drug concentrations (${\mu}M$). The compound 3d (NSC 757927) showed maximum mean percent growth inhibition (PGI) of 112.2%, while compound 3g (NSC 763374) showed less mean PGI of 40.1% in the one dose assay. The maximum anticancer activity was observed with the SR (leukemia) cell line with a $GI_{50}$ of $0.03{\mu}M$. The calculated average sensitivity of all cell lines of a particular subpanel toward the test agent showed that all the curcumin analogues showed maximum activity on leukemia cell lines with $GI_{50}$ values between 0.23 and $2.67{\mu}M$.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1849-1855
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• 2015

Staphylococcus aureus plays an important role in sepsis, septic shock, pneumonia, and wound infections. Here, we demonstrate that Lactobacillus plantarum extracts inhibited S. aureus-induced cell death of a human epithelial cell line, HT-29. In particular, we have shown that S. aureus-induced cell death was abolished by neutralization of α-toxin, indicating that α-toxin is the major mediator of S. aureus-induced cell death. DNA fragmentation experiment and caspase assay revealed that the S. aureus-induced cell death was apoptosis. L. plantarum extracts inhibited the generation of effector caspase-3 and the initiator caspase-9 in S. aureus- or α-toxin-induced cell death. Moreover, expression of Bcl-2, an anti-apoptotic protein, was activated in L. plantarum extract-treated cells as compared with the S. aureus- or α-toxin-treated only cells. Furthermore, S. aureus-induced apoptosis was efficiently inhibited by lipoteichoic acid and peptidoglycan of L. plantarum. Together, our results suggest that L. plantarum extracts can inhibit the S. aureus-mediated apoptosis, which is associated with S. aureus spreading, in intestinal epithelial cells, and may provide a new therapeutic reagent to treat bacterial infections.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2235-2240
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• 2012

c-Src is one member of non-receptor tyrosine kinase protein family that has over expression and activation in many human cancer cells. It has been shown that c-Src is implicated in various downstream signaling pathways associated with EGFR-dependent signaling such as MAPK and STAT5 pathways. Transactivation of EGFR by c-Src is more effective than EGFR ligands. To inhibit the c-Src expression, we used c-Src antisense oligonucleotide complexed with PAMAM Denderimes. The effect of c-Src antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay. Then, the expression of c-Src, EGFR and the genes related to EGFR-depended signaling with P53 was applied by real time PCR. We used western blot analysis to elucidate the effect of antisense on the level of c-Src protein expression. The results showed, c-Src antisense complexed with PAMAM denderimers has an effective role in decrease of c-Src expression and EGFR-dependent downstream genes.

• Journal of Ginseng Research
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• 제23권2호
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• pp.99-104
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• 1999

인삼과 계피 추출물의 인체 직장암 세포(HT-29), 인체 간암 세포(HepG2)및 인체 결장암 세포(HRT-18)의 증식에 미치는 영향을 in vitro에서 확인하였다. 암세포의 배양액에서 인삼 추출물 또는 계피 추출물의 효과는 농도에 비례하여 암세포의 증식을 억제하였다. 세포증식의 억제 정도는 인삼과 계피 추출물을 단독으로 첨가한 것보다 병용하여 첨가한 경우 현저한 상승 효과를 나타내어, 낮은 인삼 농도에서도 HT-29, HepG2및 HRT-18암세포의 증식을 효과적으로 억제하였으며 심지어는 사멸시켰다. 인삼과 계피의 혼합물은 세포주기 중 G1 단계에서 S단계로의 진행을 지체시킴으로써 암세포의 증식을 억제하는 것으로 나타났다.