• Radiation Oncology Journal
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• 제28권3호
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• pp.147-154
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• 2010

목 적: 대장암 세포에서 epidermal growth factor receptor (EGFR) 저해제인 nimotuzumab에 의한 방사선 민감도 증진 효과를 살펴보고자 한다. 대상 및 방법: 총 4종류의 인간 유래 대장암 세포주인 HCT-8, LoVo, WiDr, HCT-116를 nimotuzumab과 방사선을 병합 처리한 후 세포증식, 생존율, 세포주기 진행에 미치는 영향을 MTT, clonogenic survival assay, flow cytometry와 western blot을 통해 분석하였다. 결 과: 대장암 세포주에서 nimotuzumab에 의해 EGFR 인산화가 억제됨을 확인하였고 이러한 조건에서 nimotuzumab이 HCT-116을 제외한 나머지 3종류의 대장암 세포주의 방사선 민감도를 증진시킴을 확인하였다. 반면에, nimotuzumab은 방사선 조사와 무관하게 대장암 세포의 증식이나 세포 주기에는 아무런 영향을 미치지 않았다. 결 론: Nimotuzumab은 EGFR에 의한 세포 생존 신호 전달을 억제함으로써 대장암 세포의 방사선에 대한 민감도를 증가시켰다. 본 연구는 대장암의 방사선 치료에 EGFR 특이적 저해제인 nimotuzumab의 임상 적용 근거를 제공하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7291-7296
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• 2014

Different plant parts of Flacourtia indica have long been used in Ayurvedic medicine. Previous studies have demonstrated that the methanolic extract of F. indica possess anti-inflammatory properties. The present study was aimed at investigating the anticancer effects of methanol extract of Flacourtia indica (FIM) aerial parts in human colon cancer (HCT116) cells. Treatment of cells with FIM at a concentration of $500{\mu}g/ml$ for 24 hours significantly reduced cell viability and induced apoptosis, which was associated with the increased cytoplasmic expression of cytochrome c, activation of caspase-3, and the cleavage of poly-(ADP-ribose) polymerase. Incubation with FIM also inhibited the levels of Bcl-2, Bcl-xl and survivin, which are the markers of cell proliferation, whereas the expression of Bax remained unchanged. Treatment with FIM led to the generation of reactive oxygen species (ROS) in a concentration-dependent manner. Pharmacological inhibition of ROS generation by pretreatment of cells with N-acetyl cysteine abrogated FIM-induced apoptosis in HCT116 cells. Thus, these results demonstrate that FIM has anti-proliferative and pro-apoptotic effects in HCT116 cells and the effects are, at least in part, due to the ROS dependent activation of caspases.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.465-472
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• 2020

Colorectal cancer (CRC) is one of the most malignant type of cancers and its incidence is steadily increasing, due to life style factors that include western diet. Abnormal activation of canonical Wnt/β-catenin signaling pathway plays an important role in colorectal carcinogenesis. Therefore, targeting Wnt/β-catenin signaling has been considered a crucial strategy in the discovery of small molecules for CRC. In the present study, we found that Nodosin, an ent-kaurene diterpenoid isolated from Isodon serra, effectively inhibits the proliferation of human colon cancer HCT116 cells. Mechanistically, Nodosin effectively inhibited the overactivated transcriptional activity of β-catenin/T-cell factor (TCF) determined by Wnt/β-catenin reporter gene assay in HEK293 and HCT116 cells. The expression of Wnt/β-catenin target genes such as Axin2, cyclin D1, and survivin were also suppressed by Nodosin in HCT116 cells. Further study revealed that a longer exposure of Nodosin induced the G₂/M phase cell cycle arrest and subsequently apoptosis in HCT116 cells. These findings suggest that the anti-proliferative activity of Nodosin in colorectal cancer cells might in part be associated with the regulation of Wnt/β-catenin signaling pathway.

• 한방비만학회지
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• 제13권1호
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• pp.24-32
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• 2013

Objectives: The aim of this study is to investigate anti-imflammatory and protective effect for intestinal epithelial cells with Atractylodes macrocephae (AM), a traditional Korean Herbal medicine and fermented Atractylodes macrocephae (FAM) with Lactobacillus plantarum. Methods: HCT-116 and Raw 264.7 cells were used in this study. Using NO assay, we measured lipopolysaccharide (LPS)-induced anti-inflammatory effect. We measured permeability of intestinal epithelial cells with transepithelial electrical resistance and horseradish peroxide flux assay. Water soluble tetrazolium salt assay was used to see cell proliferation. All the results were presented in mean and standard deviation. We used Student's t-test for analyzing significance of results. Results: In Raw 264.7 cells NO production decreased 22.4% with pre-treatment of AM and FAM, especially with FAM in high concentration. In HCT-116 cells LPS-induced intestinal permeability had a protective effect with both AM and FAM, which was also tend to be proportional to the concentration. Cell viability increased up to 135.52% after treatment of high concentration of FAM in HCT-116, while there was no significant change in Raw 264.7 cells with herb treatments. Conclusions: These results show evidence that AM, especially fermented ones, significantly reduced intestinal membrane permeability. They also had a protective effect as well as an anti-inflammation effect for HCT-116 and Raw 264.7 cells. This suggest that FAM may be a therapeutic agent for Leaky gut syndrome by reducing intestinal permeability.

• 대한한의학방제학회지
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• 제14권2호
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• pp.21-32
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• 2006

The purpose of this study was to investigate the effect of Imyosan on apoptosis in human colorectal cancer HCT116 cells. Phellodendron amurense Rupr. and Atratylodes lancea D.C. compose Imyosan. First of all, to study the cytotoxic effect of methanol extract of Imyosan (IMS-MeOH) on HCT116 cells, the cells were treated with various concentrations of IMS-MeOH and then cell viability was determined by XTT reduction method. IMS-MeOH reduced viability of HCT116 cells in a dose and time-dependent manner. To confirm the induction of apoptosis, the c1eavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of caspase-3, procaspase-8 and procaspase-9 were examined by western blot analysis. IMS-MeOH decreased procaspase-3, procaspase-8 and procaspase-9 levels in a dose-dependent manner and induced the clevage of PARP. IMS-MeOH triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome c from mitochondria to cytosol. Furthermore, IMS-MeOH also downregulated the anti-apoptotic Bcl-2 and upregulated the pro-apoptotic-Bax. Therefore, these results suggest that IMS-MeOH induced HCT1l6 cell death through the mitochondrial pathway. To explore whether the activities of caspases was required for induction of apoptosis by IMS-MeOH, caspase-3, -8, -9 activity measured by using substrates, respectively. IMS-MeOH increased caspase-3, -8, -9 activity. Co-treatment with inhibitors of caspase-3, -8, -9 and IMS-MeOH significantly blocked IMS-MeOH-triggered apoptosis in HCT1l6 cells. These results suggest that IMS-MeOH induces caspases-mediated apoptosis.

• 한국방사선학회논문지
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• 제10권6호
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• pp.387-393
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• 2016

종양을 효율적으로 적출, 제거하기 위해서는 부수적으로 방사선 치료 및 항암화학요법을 이용하여 수술 전이나, 수술 후 종양의 크기를 줄이거나 작게 할 수는 있었으나, 종양을 적출하거나 제거하는 방법으로 외과적 절제수술요법이 가장 재래적인 수술 방법이다. 종양의 크기를 줄이거나 작게 하는 방법과, 종양을 사멸시킬 수 있는 항암화학요법은 방사성 감수성을 증가시키는 항암약제를 종양세포에 선택적으로 투여하여 방사선에 대한 감수성에 민감하게 반응 할 수 있도록 한 것이며, 다양한 생물학적인 세포증식억제 방법 중 TRAIL은 단백질을 변형 시킬 수 있으며 단백질 구조를 변형시켜 세포의 사멸에 일조를 하는 것으로 알려져 있다. 본 논문에서는 HCT-116세포를 암세포로 간주하여 TRAIL과 방사선과의 상호관계를 분석하였다. 실험결과 TRAIL과 방사선의 단독사용은 대조군과 비교해 본 결과 각각 세포증식과 세포자멸사에 유의적인 영향을 끼치지 않은 것으로 나타났다. 반대로 TRAIL로 처리하고, 방사선 조사를 병행해서 처리한 경우 HCT-116세포가 유의적으로 세포자멸사가 발생되었음을 알 수 있었고, G1대비 G0의 비율도 증가한 것으로 나타났다. 결론적으로 TRAIL은 방사선 방어적인 세포의 세포자멸사를 증가시켜 방사선 감수성을 증가시켰음을 알 수 있었으며, 또한 세포주기를 변화시켜 세포 증식 능력을 점진적으로 감소시킬 수 있었다. TRAIL은 세포자멸사를 증가시키고 세포증식 능력을 감소시켜 방사선 증감제로서 사용이 가능하다는 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1654-1663
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• 2014

To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.

• 한국지역사회생활과학회지
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• 제26권1호
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• pp.103-113
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• 2015

This study examines the effects of Korean Crataegi fructrus(KCF) and Chinese Crataegi fructrus(CCF) on the antioxidative activity and antiproliferation of human cancer cells(HCT-116 human colon, Hep G2 human liver, and A549 human lung cancer cells). The total polyphenol and flavonoid contents, and antioxidative index of the Crataegi fructrus ethanol extracts were significantly higher in KCF than in CCF. The DPPH radical-scavenging activity of the KCF ethanol extract was 82.26%(1000 ppm), and that of the CCF ethanol extract was 77.64%. Antiproliferation effects of 80% ethanol extracts of KCF and CCF on human cancer cells(HCT-116, Hep G2 and A549) increased in a dose-dependent manner. Inhibitory effects of KFC on HCT-116 and A549 cells were greater than those of CCF. The results suggest that ethanol extracts of Crataegi fructrus have antioxidative and hyperplasia inhibition effects on human cancer cells.

• 약학회지
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• 제55권5호
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• pp.391-397
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• 2011

KML001 reduced the proliferation of HCT116 cells in a concentration- and time-dependent manner without change of cell viability. Beclin-1 expression was significantly attenuated by KML001 (P<0.05), but no significant changes were observed in KML001-treated cells. The number of cells in G1 phase was increased 48 hr after KML001 treatment. Furthermore, a dramatic reduction in the frequency of beating and the number of embryoid bodies of the cells was noted after treatment. Taken together, KML001 suppresses the proliferation of HCT116 cells, which might be due to G1 phase arrest.

• 대한본초학회지
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• 제21권2호
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• pp.37-46
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• 2006

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.