• Title/Summary/Keyword: cell growth assay

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Evaluation of Anticancer Activity of Curcumin Analogues Bearing a Heterocyclic Nucleus

  • Ahsan, Mohamed Jawed
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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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 $2.67{\mu}M$.

Antineoplastic Effect of Several Herbal Medicines on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (황기, 당귀, 칠피, 천화분의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun Soo;Lee, Min Hye;Ko, Seong Gyu;Choi, You Kyung;Jun, Chan Young;Park, Jong Hyeong
    • Journal of Society of Preventive Korean Medicine
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    • v.18 no.1
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    • pp.83-92
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    • 2014
  • Objective : This study was performed to investigate the antineoplastic effect of Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes and Trichosanthes kirilowii on SNU-80 anaplastic thyroid carcinoma cell line. Method : We examined whether our herbal medicines decreases cell growth rate of SNU-80 using MTT assay. We performed western blot analysis to verify that our herbal medicines induces apoptosis via caspase-dependent mechanism. We also performed wound healing assay and transwell invasion assay to investigate whether our herbal medicines affects the migration and invasion of anaplastic cancer cells, SNU-80. We also carried out ELISA assay to know our herbal medicines suppresses the expression of proinvasive molecules, such as VEGF and MMP-2 secreted from SNU-80. Results : MTT assay demonstrates that Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed strongly the growth of SNU-80. Western blot analysis demonstrates that Trichosanthes kirilowii induces apoptosis activating the cleavages of caspases (caspase-8, caspase-3) and PARP. Wound healing assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the migration of SNU-80. Transwell invasion assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the invasion of SNU-80. Elisa assay demonstrates that Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed the expression of VEGF and MMP-2. Conclusion : We could conclude that several herbal medicines suppresses the growth and inhibits the migration and invasion of SNU-80 which is anaplastic thyroid cancer cells. Especially, Rhus verniciflua Stokes, Trichosanthes kirilowii had stronger anti-cancer effect suggesting that we can apply them to treat anaplastic thyroid cancer.

Rat Gonadotropes and Somatotropes Express Growth Hormone Releasing Hormone Gene in the Pituitary (흰쥐 뇌하수체 Gonadotropes와 Somatotropes에서의 Growth Hormone Releasing Hormone 유전자 발현)

  • 이성호
    • Development and Reproduction
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    • v.2 no.2
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    • pp.189-196
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    • 1998
  • Several lines of evidence indicate that some neuropeptides classically associated hypothalamus have been found in pituitary gland, suggesting the existence of local regulation of pituitary function. Among the hypothalamic releasing hormones, genes for TRH and GnRH are expressed in the rat anterior pituitary gland. The present study was carried out to investigate the expression of the GHRH gene in rat anterior pituitary and the pituitary-derived cell lines. The presence of GHRH transcripts in pituitary tissue was shown by 3'rapid amplification of cDNA end (3'-RACE) analysis. In reverse transcription-polymerase chain reaction (RT-PCR) study, GHRH cDNA fragments were amplified from two pituitary-derived cell lines, $\alpha$T3 cells originated from mouse gonadotrope and GH3 cells from rat somatolactotrope. Immunoreactive GHRH was detected in large and medium-sized pituitary cells by immunocytochemistry. Significant amounts of GHRH-like molecules were found in the GH3 cell extracts. In RNase protection assay, the level of pituitary GHRH mRNA was augmented by ovariectomy. These results demonstrate that GHRH gene is expressed in the rat gonadotropes and somatotropes, and suggest that the pituitary GHRH could be participated in the paracrine and/or autocrine regulation of cell proliferation, as well as promoting growth hormone secretion.

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Growth Inhibition of Extract from Sulfur fed Duck Carcass against Various Cancer Cell Lines (유황오리 추출물의 각종 암세포에 대한 생육억제 효과)

  • 최귀헌;김창한
    • Food Science of Animal Resources
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    • v.22 no.4
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    • pp.348-351
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    • 2002
  • This study was carried out to investigate the anticancer effect of extracts from sulfur fed duck carcass. Growth inhibition of cancer cell lines was measured by MTT assay. Eleven cancer cell lines, such as Calu-3(human lung carcinoma), SK-MES-1(human lung carcinoma), HL6O(human leukemia), KB(human epidermoid of mouth carcinoma), Farrow(human melanoma), HEP-2(human larynx carcinoma), SNU-1(human stomach carcinoma), K-562 (human leukemia), WiDr(human colon carcinoma), P388(mouse leukemia) and 3LL(mouse lung carcinoma) showed the growth inhibition higher than 50%, but those, such as SF-188(human brain carcinoma), A-549(human lung carcinoma) and HEC-lB(human uterus carcinoma) showed the growth inhibition lower than 50% in the extract of sulfur fed duck carcass at the concentration of 10 mg/㎖. The sulfur fed duck carcass extract had better growth inhibition than the normal counterpart against various cancer cell lines at the concentration of 10 mg/㎖. When the effect of growth inhibition of an effluent by different concentrations of methyl alcohol(25, 50, 75 and 100%) tested on Diaion HP-20 column chromatography, an effluent by concentration of 100% methyl alcohol showed the most strong effect of growth inhibition against HEP-2(human larynx carcinoma).

Growth Inhibition of Human Head and Neck Squamous Cell Carcinomas by Angelica decursiva Extracts

  • Shin, Woo-Cheol;Kim, Chun-Sung;Kim, Heung-Joong;Lee, Myoung-Hwa;Kim, Hye-Ryun;Kim, Do-Kyung
    • International Journal of Oral Biology
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    • v.35 no.4
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    • pp.153-158
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    • 2010
  • Angelica decursiva has been used in Korean traditional medicine as an antitussive, an analgesic, an antipyretic and a cough remedy. However, the anti-cancer properties of Angelica decursiva have not yet been well defined. In our current study the cytotoxic activity of ethanol extracts of Angelica decursiva root (EEAD) and the mechanism of cell death exhibited by EEAD were examined in FaDu human head and neck squamous cell carcinoma cells. The cytotoxic effects of EEAD upon the growth of FaDu cells were examined with an MTT assay. In addition, the mechanism of cell death induced by EEAD was evaluated by DNA fragmentation analysis, immunoblotting and caspase activation measurements. EEAD induced apoptotic cell death in FaDu cells in a concentration- and time-dependent manner, as determined by MTT assay and DNA fragmentation analysis. Furthermore, the proteolytic processing of caspase-3, -7 and -9 was increased by EEAD treatment of FaDu cells. In addition, the activation of caspase-3 and -7 was detected in living FaDu cells by fluorescence microscopy. These results suggest that EEAD can induce apoptosis and suppress cell growth in cancer cells and may have utility as a future anticancer therapy.

Antitumor Effects of Kluyveromyces marxianus TFM-7 Isolated from Kefir

  • Lee, Hyun-Jung;Nam, Bo-Ra;Kim, Jin-Man;Kim, Ji-Yeon;Paik, Hyun-Dong;Kim, Chang-Han
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.133-137
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    • 2007
  • The Strain TFM-7, Which has an antitumor effect, was isolated from Kefir and identified based on analysis using the API 50 CHL kit and 265 rDNA sequencing. Strain TFM-7 was confirmed to belong to the genus Kluyveromyces. Analysis of the 265 rDNA nucleotide sequences found strain TFM-7 to be related to Kluyveromyces marxianus. NRRL Y-828IT. K. marxianus. TFM-7 was cultured with potato dektrose broth medium at $27^{\circ}C$ for 72 hr, and its inhibition effects on the proliferation of seven tumor cell lines and a normal cell line were assessed using the MTT assay. The antitumor effects and growth characteristics of K. marxianus TFM-7 were investigated during a culture period of 7 days. By the $3^{rd}\;day$, K. marxianus TFM-7 showed a dry cell weight 2.39 g/L, a pH of 4.39, an ethanol content of 0.89%, and an inhibition effect on the proliferation of seven tumor cell lines above 50%, except for A-549 tumor cell line. K. marxianus TFM-7 was the most effective at inhibiting the growth of Hep-2 cell line among all tumor cell lines tested. Growth inhibition of a normal cell line, NIH/3T3, was less than 35%, suggesting a decreased level of cytotoxicity toward normal cells. These results indicate that K. marxianus TFM-7 may have used as a yeast strain with antitumor activity.

The Effects of Gokgisaeng on Anti-inflammation and Rat C6 Glioma Cell Migration (곡기생(槲寄生)의 항염증 효능 및 암세포 이주저해에 미치는 영향)

  • Kim, Hyun-Young;Jang, Soo-Young;Jeong, Ji-Cheon;Shin, Hyeon-Cheol
    • The Journal of Internal Korean Medicine
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    • v.34 no.1
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    • pp.31-45
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    • 2013
  • Objectives : Gokgisaeng (Korean mistletoe) is used for the treatment of inflammatory and cancer diseases in traditional Korean medicine and its major component lectins have been reported to induce nitric oxide (NO) in RAW 264.7 macrophages, and also induce apoptosis of various types of cancer cells, although its modulatory effects on cancer cell migration and macrophage activation is poorly understood. The aim of this study is to clarify molecular mechanisms of action responsible for the anti-inflammatory and antitumor migration potentials of Korean mistletoe extract (KME). Methods : We investigated the anti-inflammatory activity of KME on NO production and inducible nitric oxide synthase (iNOS) expression by lipopolysaccharide (LPS) in both RAW 264.7 macrophages and rat C6 glioma cells, and also evaluated inhibitory efficacy on glioma cell growth and migration. For assessment, XTT assay, nitrite assay, RT-PCR, scratch-wound and Boyden chamber assay, and western blot analysis were performed. Results : Previously reported, unlike the efficacy of Gokgisaeng lectin, KME inhibited NO production and iNOS expression, and suppressed pro-inflammatory mediators including IL-$1{\beta}$, IL-6, COX-2, iNOS in LPS-stimulated RAW 264.7 cells. Furthermore, KME suppressed tumor cell growth and migration, and it also inhibited LPS-induced NO release and iNOS activation by down-regulating expression of protein kinase C (PKC) and phosphorylation of ERK in C6 glioma cells. Conclusions : Our research findings provide evidence that KME can play a significant role in blocking pro-inflammatory reaction and malignant progression of tumors through the suppression of NO/iNOS by down-regulating of inflammatory signaling pathways, PKC/ERK.

Mouse Melanoma Cell Migration is Dependent on Production of Reactive Oxygen Species under Normoxia Condition

  • Im, Yun-Sun;Ryu, Yun-Kyoung;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.20 no.2
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    • pp.165-170
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    • 2012
  • Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

Study on Antitumor Activity of Samiyeongeontanggamibang(SYTG) (사미연견탕가미방(四味軟堅湯加味方)이 항암(抗癌) 및 항전이(抗轉移) 효과(效果)에 미치는 영향)

  • Bae, Moon-Yong;Kang, In-Cheol;Kim, Sung-Hoon
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.5 no.1
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    • pp.33-46
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    • 1999
  • To evaluate the antitumor and antimetastatic effects of Samiyeongeontanggamibang(SYTG), We have examined whether SYTG can inhibit the growth of several tumor cell lines, tumor cell adhesion, experimental tumor metastasis and increase survival rate of tumor-bearing mice by inhibition of DNA topoisomrase activity. The results were obtained as follows: 1. SYTG extracts revealed an efficient cytotoxicity against A549, SK-OV-3, B16-F10, and SK-MEL-2 cell lines. 2. SYTG extracts inhibited DNA topo-isomerase I from calf thymus. 3. The T/C% in S-180 bearing ICR mice treated with SYTG was 115.8% 4. SYTG extracts significantly inhibited adhesion of A549 cell to complex extracellular matrix. 5. In pulmonary colonization assay, SYTG suppressed lung metastases in tumor cell-injected mice. 6. In CAM assay, SYTG extracts inhibited angiogenesis at $15{\mu}g/egg$ concentration as compared with control. These results suggested that SYTG extracts might be a potent inhibitor of cancer.

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AN EXPERIMENTAL STUDY ON THE STIMULATORY EFFECTS OF EPIDERMAL GROWTH FACTOR AND TRANSFORMING GROWTH FACTOR-α ON THE GROWTH OF SQUAMOUS CANCER CELL LINES (Epidermal Growth Factor 와 Transforming Growth Factor-α가 인체 구강편평상피세포암 세포의 성장에 미치는 영향에 관한 실험적 연구)

  • Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.20 no.4
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    • pp.334-340
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    • 1998
  • Stimulatory effects of epidermal growth factor (EGF) and transforming growth $factor-{\alpha}$($TGF-{\alpha}$) on the growth of squamous cancer cell lines established from human oral cancer tissue with moderate differentiation were studied in vitro. After culturing in serum-free media for 24 hours, growth factors-EGF only, $TGF-{\alpha}$ only and EGF, $TGF-{\alpha}$ together-were added to the media and numbers of cells were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and compared with the control at 96, 144 hours. Each of EGF and $TGF-{\alpha}$ showed statistically significant stimulatory effects on the growth of cells respectively. Dose-dependent relationship of the stimulatory effects were not clearly demonstrated. The effects of EGF were higher than those of $TGF-{\alpha}$ and combinative administration showed higher effects than those of single uses. In conclusion, EGF may play an important and major role in differentiation and growth of human oral squamous cancer cells. $TGF-{\alpha}$, produced from cells activated by EGF, also can stimulate the cell growth and could be an alternative ligand for EGF receptor.

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