• Title/Summary/Keyword: cell growth assay

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The effects of Injinchunggan-tang on Cell Growth and Apoptosis in Human Hepatic Stellate Cell Line LX2 (인진청간탕(茵蔯淸肝湯)이 간성상세포의 세포성장과 사멸에 미치는 영향)

  • Kim, Sang-Joo;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.32 no.4
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    • pp.519-529
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    • 2011
  • Objectives : This study was performed to investigate the effects of Injinchunggan-tang on cell growth and apoptosis in human hepatic stellate cell line LX2. Materials and Methods : Hepatic stellate cells were treated with various concentrations of Injinchunggan-tang extract for 24, 48 and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, apoptosis, caspase activity, caspase inhibitor and the mRNA of the Bcl-2, and Bax with ${\beta}$-actin were measured by using MTT assay, apoptosis assay and RT-PCR. Results : Proliferation, and mRNA expression of the hepatic stellate cells were inhibited by Injinchunggan-tang treatment in a dose-dependent manner. This indicates the prescription has inhibitory effect on fibrogenesis of the liver by regulating the fibrogenesis-associated genes in transcription. Cell viability was inhibited in time- and dose-dependent manners. Conclusions : These results suggest that Injinchunggan-tang would be beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis.

Comparative Studies of Adriamycin and 28-Deacetyl Sendanin on In Vitro Growth Inhibition of Human Cancer Cell Lines

  • Kim, Hwan-Mook;Oh, Goo-Taeg;Han, Sang-Bae;Hong, Dong-Ho;Hwang, Bang-Yeon;Kim, Young-Ho;Lee, Jung-Joon
    • Archives of Pharmacal Research
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    • v.17 no.2
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    • pp.100-103
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    • 1994
  • The limonoid compound (28-deacetyl sendanin0 isolated from the fruit of Melia toosendan SIEB. et ZUCC. was evaluated on anticancer activity. According to a standard in vitro cytotoxicity assy, eight human cancer cell lines and SRB assay were introduced for present evaluation. As a positive standard, adriamycin was tested in parallel. The cell lines were originated from six different organs. In view of dose-response profiles to 28-deacetyl sendanin, the most sensitive cells were SF-539 and PC-3 which were derived from CNS and prostate, respecitively. In contrast, all the cell lines responded similarly to adriamycin to give rise to nearly indentical six cell lines were more sensitive to 28-deacetyl sendanin and two were more resistant. As a result, 28-deacetyl sendanin had more senstive and selective inhibitory effects on in vitro growth of human cancer cell lines in a comparison with adriamycin.

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Short-Hairpin RNA-Mediated MTA2 Silencing Inhibits Human Breast Cancer Cell Line MDA-MB231 Proliferation and Metastasis

  • Lu, Jun;Jin, Mu-Lan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.14
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    • pp.5577-5582
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    • 2014
  • Objective: To observe the effects of metastasis-associated tumor gene family 2 (MTA2) depletion on human breast cancer cell proliferation and metastasis. Methods: A short-hairpin RNA targeting MTA2 was chemically synthesized and transfected into a lentivirus to construct Lv-shMTA2 for infection into the MDA-MB231 human breast cancer cell line. At 48 hours after infection cells were harvested and mRNA and protein levels of MTA2 were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Cell viability and metastasis were assessed by CCK-8, wound-healing assay and Transwell assay, respectively. In addition, a xenograft model of human breast cancer was constructed to investigate cancerous cell growth and capacity for metastasis. Results: After infection with Lv-shMTA2, mRNA and protein levels of MTA2 was significantly reduced (p<0.05) and MDA-MB231 cell proliferation and metastasis were inhibited (p<0.05). In addition, mean tumor size was smaller than that in control group nude mice (p<0.05) and numbers of metastatic deposits in lung were lower than in control group mice (p<0.05). Depletion of MTA2 affected MMP-2 and apoptosis-related protein expression. Conclusions: For the first time to our knowledge we showed that MTA2 depletion could significantly inhibit human breast cancer cell growth and metastasis, implying that MTA2 might be involved in the progression of breast cancer. The role of MTA2 in breast cancer growth and metastasis might be linked with regulation of matrix metalloproteinase and apoptosis.

Effects of Gagamgilgyung-tang on the Proliferation and Apoptosis of Human Lung Cancer Cell (가감길경탕이 인체 폐암세포의 증식 및 사멸에 미치는 영향에 관한 연구)

  • 이충섭;정희재;신순식;정승기;이형구
    • The Journal of Korean Medicine
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    • v.23 no.1
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    • pp.24-36
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    • 2002
  • Objectives: The chemotherapeutic potential of Gagamgilgyung-tang for the treatment of human lung cancer, the antitumorigenic effects of Gagamgilgyung-tang on the proliferation and apoptosis of human lung cancer cell line A427 were investigated using molecular biological approaches, Methods: To determine Gagamgilgyung-tang concentrations which do not evoke cytotoxic damage to the cell line, cell viability was examined by MTT assay. To prove Gagamgilgyung-tang's antitumorigenic potential to human lung cancer, [3H]thymidine incorporation assay, trypan blue exclusion and Cpp32 protease activity assays and quantitative RT-PCR analysis were examined. Results: While A427 cells treated with $0.1-2.0{\mu\textrm{g}}/ml$ of Gagamgilgyung-tang showed no recognizable effect, marked reductions of cell viability were detected at concentrations over $5.0{\;}\mu\textrm{g}/ml$. DNA replication of A427 cells was inhibited by Gagamgilgyung-tang in a dose-dependent manner and Gagamgilgyung-tang induced the G1 cell cycle arrest through inhibition of DNA replication. Gagamgilgyung-tang triggered apoptotic cell death of A427 and enhanced the apoptotic sensitivity of the cells that were injured by a DNA damage-inducing chemotherapeutic drug etoposide. Gagamgilgyung-tang induces expression of growth-inhibiting genes such as p53 and p21/Wafl whereas it inhibited expression of growth-promoting genes such as c-Myc and Cyclin D1. Expression of a representative apoptosis-inducing gene Bax was also found to be induced by Gagamgilgyung-tang while apoptosis-suppressing Bcl-2 expression was not changed. Conclusions: Gagamgilgyung-tang could suppress the abnormal growth of tumor cells by suppressing the survival of genetically altered cells via induction of apoptosis. This study suggests that Gagamgilgyung-tang might have an antitumorigenic potential to human lung cancer cells, which might be associated with its growth-inhibiting and apoptosis-inducing properties.

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Murrayafoline A Induces a G0/G1-Phase Arrest in Platelet-Derived Growth Factor-Stimulated Vascular Smooth Muscle Cells

  • Han, Joo-Hui;Kim, Yohan;Jung, Sang-Hyuk;Lee, Jung-Jin;Park, Hyun-Soo;Song, Gyu-Yong;Nguyen, Manh Cuong;Kim, Young Ho;Myung, Chang-Seon
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.5
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    • pp.421-426
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    • 2015
  • The increased potential for vascular smooth muscle cell (VSMC) growth is a key abnormality in the development of atherosclerosis and post-angioplasty restenosis. Abnormally high activity of platelet-derived growth factor (PDGF) is believed to play a central role in the etiology of these pathophysiological situations. Here, we investigated the anti-proliferative effects and possible mechanism(s) of murrayafoline A, a carbazole alkaloid isolated from Glycosmis stenocarpa Guillamin (Rutaceae), on PDGF-BB-stimulated VSMCs. Murrayafoline A inhibited the PDGF-BB-stimulated proliferation of VSMCs in a concentration-dependent manner, as measured using a non-radioactive colorimetric WST-1 assay and direct cell counting. Furthermore, murrayafoline A suppressed the PDGF-BB-stimulated progression through $G_0/G_1$ to S phase of the cell cycle, as measured by [$^3H$]-thymidine incorporation assay and cell cycle progression analysis. This anti-proliferative action of murrayafoline A, arresting cell cycle progression at $G_0/G_1$ phase in PDGF-BB-stimulated VSMCs, was mediated via down-regulation of the expression of cyclin D1, cyclin E, cyclin-dependent kinase (CDK)2, CDK4, and proliferating cell nuclear antigen (PCNA), and the phosphorylation of retinoblastoma protein (pRb). These results indicate that murrayafoline A may be useful in preventing the progression of vascular complications such as restenosis after percutaneous transluminal coronary angioplasty and atherosclerosis.

Cytotoxic Effect of Korean Traditional Prescriptions on the Human Gastric Cancer Cell Lines (한약처방제의 인체 위암 세포주에 대한 세포독성 효과에 관한 연구)

  • Kim, Eun-Hae;Eun, Young-Ah;Kang, Bong-Joo;Sung, Hyun-Jae;Park, Kap-Joo
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.233-238
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    • 1997
  • ln order to search for antigastric cancer agents from Korean traditional prescriptions. We selected 41 traditional prescriptions, based on a review of the Korean traditional medicine books. Both boiling water and methanol extracts were tested, by means of the Sulforhodamine B (SRB) protein assay. Six of the 41 water extracts; #3, #34, #35, #38, #40, #41 showed efficacy against gastric cancer cell (AGS: Human gastric carcinoma, ATCC HTB 103). #3 inhibited 50% cancer cell growth1 at the concentration of $152\;{\mu}g/ml$, #34, #35, #38, #40 and #41 inhibited 50% cancer cell growth at the concentration of $145\;{\mu}g/ml$, $129\;{\mu}g/ml$, $173\;{\mu}g/ml$, $10\;{\mu}g/ml$ and $19\;{\mu}g/ml$ respectively. Ten of the 41 methanol extracts; #1, #3, #32, #33, #35, #36, #37, #38, #41 were active. #1 inhibited 50% cancer cell growth at the concentration of $206\;{\mu}g/ml$, #3, #32, #33, #35, #36, #37, 738, #40, #41 inhibited 50% cancer cell growth at the concentration of $133\;{\mu}g/ml$, $159\;{\mu}g/ml$, $199\;{\mu}g/ml$, $147\;{\mu}g/ml$, $113\;{\mu}g/ml$, $187\;{\mu}g/ml$, $130\;{\mu}g/ml$, $9\;{\mu}g/ml$, $15\;{\mu}g/ml$ respectively. Prescription #3, #35, #38, #40, #41 were also interesting because both methanol and water extracts were active.

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The Study of Aati-cancer Effects of Bee Venom for Aqua-acupuncure (약침용(藥鍼用) 봉독성분(蜂毒成分) 중(中) Apamin, Melittin의 항암작용(抗癌作用))

  • Kwon, Do-Hee;Lee, Jae-dong;Choi, Do-Yong
    • Journal of Acupuncture Research
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    • v.18 no.1
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    • pp.129-145
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    • 2001
  • Objectives : To characterize the antitumorigenic potential of three representative bee venom components, Melittin, Apamin, and Phospholipase A2, their effects on cell proliferation and apotosis of the human melanoma cell line SK-MEL-2 were analyzed using molecular biological approaches. Methodes & Results : To determine the doses of the drugs that do not induce cytotoxic damage to this cell line, cell viability was examined by MTT assay. While SK-MEL-2 cells treated with 0.5 - 2.0㎍/㎖ of each drug showed no recognizable cytotoxic effect, marked reductions of cell viability were detected at concentrations over 5.0㎍/㎖. [3H]thymidine incorporation assay for cell proliferation demonstrated that DNA replication of SK-MEL-2 cells is inhibited by Apamin and Phospholipase A2 in a dose-dependent manner. Consistent with this result, the cells were accumulated at the G1 phase of the cell cycle after treatment with Apamin and Phospholipase A2, whereas no detectable change in cell proliferation was identified by Melittin treatment. In addition, tryphan blue exclusion and flow cytometric analyses showed that all of these drugs can trigger apoptotic cell death of SK-MEL-2, suggesting that Melittin, Apamin, and Phospholipase A2 have antitumorigenic potential through the suppression of cell growth and/or induction of apoptosis. Qantitative RT-PCR analysis revealed that Apamin and Phospholipase A2 inhibit expression of growth-promoting genes such as c-Jun, c-Fos, and Cyciin D1. Furthermore, Phospholipase A2 induced tumor suppressors p53 and p21/Wafl. In addition, all three drugs were found to activate expression of a representative apoptosis-inducing gene Bax while expression of apoptosis-suppressing Bcl-2 and Bcl-XL genes was not changed. Taken together, this study strongly suggests that Metittin, Apamin, and Phosphalipase A2 may have antitumorigenic activities, which are associated with its growth-inhibiting and/or apoptosis-inducing potentials.

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Suppression of MCF-7 Breast Cancer Cell Multiplication by Alismatis rhizoma Extract

  • Da Hyun Kim;Eun Ju Yang;Jeong Hyun Chang
    • Biomedical Science Letters
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    • v.30 no.3
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    • pp.137-142
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    • 2024
  • The aim of this study was to investigate the inhibitory effects of Alismatis rhizoma extract on breast cancer cell growth and elucidate the underlying mechanisms. The growth inhibitory effects of Alismatis rhizoma extract on breast cancer cells were measured using the WST-1 assay, while its impact on relevant proteins and genes was analyzed using real-time polymerase chain reaction. The results demonstrated significant inhibition of breast cancer cell growth by Alismatis rhizoma, with the inhibitory effect showing a positive correlation with the dosage, treatment duration, and quantity. Furthermore, Alismatis rhizoma extract markedly decreased the expression levels of cell cycle protein D1 and suppressed cell migration and invasion. In conclusion, Alismatis rhizoma exhibits the potential to inhibit breast cancer cell growth, possibly by regulating cell cycle progression and inhibiting cell migration and invasion. These findings suggest that Alismatis rhizoma extract possesses anticancer properties against human breast cancer cells. Further investigation of its reuglation of action will provide foundational data, potentially paving the way for its development as an anticancer therapeutic agent.

ANTI-TUMOR EFFECTS OF VASCULAR ENDOTHELIAL GROWTH FACTOR INHIBITOR ON ORAL SQUAMOUS CELL CARCINOMA CELL LINES (혈관내피세포성장인자 억제제에 의한 구강편평상피세포암종 세포주의 성장 억제 효과)

  • Han, Se-Jin;Lee, Jae-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.2
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    • pp.66-73
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    • 2009
  • Tumor angiogenesis is a process leading to formation of blood vessels within tumors and is crucial for maintaining a supply of oxygen and nutrients to support tumor growth and metastasis. Vascular endothelial growth factor(VEGF) plays a key role in tumor angiogenesis including induction of endothelial cell proliferation, migration, survival and capillary tube formation. VEGF binds to two distinct receptors on endothelial cells. VEGFR-2 is considered to be the dominant signaling receptor for endothelial cell permeability, proliferation, and differentiation. Bevacizumab(Avastin, Genetech, USA) is a monoclonal antibody against vascular endothelial growth factor. It is used in the treatment of cancer, where it inhibits tumor growth by blocking the formation of new blood vessels. The goal of this study is to identify the anti-tumor effect of Bevacizumab(Avastin) for oral squamous cell carcinoma cell lines. Human squamous cell carcinoma cell line(HN4) was used in this study. We examined the sensitivity of HN4 cell line to Bevacizumab(Avastin) by using in vitro proliferation assays. The results were as follows. 1. In the result of MTT assay according to concentration of Bevacizumab(Avastin), antiproliferative effect for oral squamous cell carcinoma cell lines was observed. 2. The growth curve of cell line showed the gradual growth inhibition of oral squamous cell carcinoma cell lines after exposure of Bevacizumab(Avastin). 3. In the apoptotic index, groups inoculated Bevacizumab(Avastin) were higher than control groups. 4. In condition of serum starvation, VEGFR-2 did not show any detectable autophosphorylation, whereas the addition of VEGF activated the receptor. Suppression of phosphorylated VEGFR-2 and phosphorylated MAPK was observed following treatment with Bevacizumab(Avastin) in a dose-dependent manner. 5. In TEM view, dispersed nuclear membrane, scattered many cytoplasmic vacuoles and localized chromosomal margination after Bevacizumab(Avastin) treatment were observed. These findings suggest that Bevacizumab(Avastin) has the potential to inhibit MAPK pathway in proliferation of oral squamous cell carcinoma cell lines via inhibition of VEGF-dependent tumor growth.

The Comparative of Inhibitory Effect of Various Solvent Extracts from Aloe arborescens and Aloe vera on Tumor Cell Lines Using Clonoginic Assay (Clonogenic Assay를 이용한 Aloe arborescens와 Aloe vera 용매 추출물의 종양세포 억제효과의 비교)

  • Hong, Hee-Sun;Lee, Keyong-Ho;Kim, Jeong-Hwan;Kang, Hee-Gon;Cho, Choa-Hyoung;Kim, Chang-Han
    • Korean Journal of Pharmacognosy
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    • v.30 no.3
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    • pp.275-279
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    • 1999
  • The solvent extracts from Aloe arborescens and Aloe vera were randomly screened for inhibitory effects on the growth of tumor cell lines. In case of Aloe vera extracts, butyl alcohol extract and ethyl alcohol extract showed antitumor activity at $100\;{\mu}g/ml$ on lung cell lines(A427, Sk-mes-1, Calu-3 and 3LL). In Aloe arborescens extracts, butyl alcohol extract and ethyl alcohol extract exerted high activity at $100\;{\mu}g/ml$ on breast cell line(Hs-578T) and lung cell line(Sk-mes-1), respectively. The solvent extracts from Aloe vera exerted antitumor activity broadly on various tumor cell lines. In contract, the solvent extracts from Aloe arborescens exerted specific antitumoricity on a few tumor cell lines.

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