• 제목/요약/키워드: Colony forming assay

검색결과 105건 처리시간 0.024초

수치파두(修治巴豆) 및 파두가황연(巴豆加黃連)의 세포독성(細胞毒性)과 항종양(抗腫瘍) 효과(效果)에 관(關)한 실험적(實驗的) 연구(硏究) (Experimental Studies on the Change of Cytotoxic and Antitumor Effects according to the Prebrewed Method of Semen Tiglii and Rhizoma Coptidis)

  • 조성각;문구;문석재
    • 대한한방종양학회지
    • /
    • 제1권1호
    • /
    • pp.191-211
    • /
    • 1995
  • This experiment was designed to study the change of cytotoxic and antitumor effects according to the prebrewed method of Semen Tiglii and Rhizoma Coptidis. The cytotoxic and antitumor effects were evaluated on human cell lines(A 549, Caki-1, LL2, Sarcoma 180, NIH/3T3) after exposure to prebrewed Semen Tiglii and Rhizoma Coptidis water extract 0.1, 0.2, 0.4, 0.8, 1.6 mg/ml using in MTT assay, LDH, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. The cytotoxicity from the result of MTT assay was low slightly in the ST II(炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of STI(生巴豆霜). 2. The cytotoxicity from the result of LDH was low slightly in the ST Ⅱ (炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of ST I(生巴豆霜). 3. The antitumor affect on A 549 tumor cell from the result of colony forming efficiency was low slightly in the ST II (炒巴豆霜) and ST I + RC(生巴豆霜加黃連). 4. The antitumor effect on Caki-1 tumor cell from the result of SRB assay was low slightly in the ST II (炒巴豆霜). 5. Median survival time and Increased life span increased slightly in the ST I RC(生巴豆霜加黃連) and ST II (炒巴豆霜). 6. The inhibitory effect on the growth of Sarcoma 180 and Lewis lung carcinoma tumor cell increased slightly in the ST I + RC(生巴豆霜加黃連) and ST II (炒巴豆霜).

  • PDF

Mitofusin-2 enhances cervical cancer progression through Wnt/β-catenin signaling

  • Sung Yong Ahn
    • BMB Reports
    • /
    • 제57권4호
    • /
    • pp.194-199
    • /
    • 2024
  • Overexpression of mitofusin-2 (MFN2), a mitochondrial fusion protein, is frequently associated with poor prognosis in cervical cancer patients. Here, I aimed to investigate the involvement of MFN2 in cervical cancer progression and determine the effect of MFN2 on prognosis in cervical cancer patients. After generating MFN2-knockdown SiHa cells derived from squamous cell carcinoma, I investigated the effect of MFN2 on SiHa cell proliferation using the Cell Counting Kit-8 assay and determined the mRNA levels of proliferation markers. Colony-forming ability and tumorigenesis were evaluated using a colony-formation assay and tumor xenograft mouse models. The migratory and invasive abilities associated with MFN2 were measured using wound-healing and invasion assays. Wnt/β-catenin-mediated epithelial-mesenchymal transition (EMT) markers related to MFN2 were assessed through quantitative RT-PCR. MFN2-knockdown SiHa cells exhibited reduced proliferation, colony formation, migration, invasion, and tumor formation in vivo. The motility of SiHa cells with MFN2 knockdown was reduced through Wnt/β-catenin-mediated EMT inhibition. MFN2 promoted cancer progression and tumorigenesis in SiHa cells. Overall, MFN2 could serve as a therapeutic target and a novel biomarker for cervical cancer.

Detection of Bacillus Cereus Using Bioluminescence Assay with Cell Wall-binding Domain Conjugated Magnetic Nanoparticles

  • Park, Chanyong;Kong, Minsuk;Lee, Ju-Hoon;Ryu, Sangryeol;Park, Sungsu
    • BioChip Journal
    • /
    • 제12권4호
    • /
    • pp.287-293
    • /
    • 2018
  • Bacillus cereus can cause blood infections (i.e., sepsis). Its early detection is very important for treating patients. However, an antibody with high binding affinity to B. cereus is not currently available. Bacteriophage cell wall-binding domain (CBD) has strong and specific binding affinity to B. cereus. Here, we report the improvement in the sensitivity of an ATP bioluminescence assay for B. cereus detection using CBD-conjugated magnetic nanoparticles (CBD-MNPs). The assay was able to detect as few as 10 colony forming units (CFU) per mL and $10^3CFU\;per\;mL$ in buffer and blood. CBD-MNPs did not show any cross-reactivity with other microorganisms. These results demonstrate the feasibility of the ATP assay for the detection of B. cereus.

Antimicrobial activity and cytotoxicity test of Scrophularia ningpoensis hemsl extracts against Klebsiella pneumoniae

  • Yook, Keun-Dol
    • 한국컴퓨터정보학회논문지
    • /
    • 제21권5호
    • /
    • pp.135-139
    • /
    • 2016
  • Scrophularia ningpoensis hemsl has been traditionally used in China and Vietnam for treatment of bacteria, atopy, pimple, tonsillitis, angina and encephalitis for a long time. The main objectives of this study were to evaluate the antibacterial activity of the Scrophularia ningpoensis hemsl extract on biofilm formation of Klebsiella pneumoniae. Antibacterial activity was conducted using disc diffusion assay and minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were determined using the broth micro dilution method in accordance to Clinical and Laboratory Standards Institute guidelines(CLSI). Furthermore, cytotoxicity on L929 were assessed using animal cell culture for the proliferation test(MTT cell assay) and the biofilm forming capacity of the K. pneumoniae were determined using the colony forming unit (CFU) assay. The extract exhibited considerable antibacterial activity. K. pneumoniae was susceptible to the extract with the MIC and MBC of 0.1875 and $1.5mg/m{\ell}$ respectively. Cytoxicity test in L929 showed no sign of toxicity at the concentration of $0.75mg/m{\ell}$ and at the same concentration the extract caused inhibition of bacterial biofilm formation. The extract of Scrophularia ningpoensis hemsl possesses an in vitro antibacterial antibiofilm activities against K. pneumoniae, with no sign of cytoxicity on L929.

Detection of Salmonella in Milk by Polymerase Chain Reaction

  • Park, Weon-Sang
    • 한국식품위생안전성학회지
    • /
    • 제15권3호
    • /
    • pp.262-266
    • /
    • 2000
  • 우유에 포함된 Salmonella enteritidis를 효과적으로 분리하는 방법을 찾고 이를 이용하여 우유속의 S. enteritidis의 량을 추정하는 방법을 개발하였다. 일정량의 S. enteritidis를 접종한 우유로부터 guanidine thiocyanate/phenol/chloroform을 이용하여 DNA를 추출한 후 중합효소반응으로 S. enteritidis 섬모항원 유전자를 선택적으로 검출함으로써 우유 1ml당 200 colony forming unit까지 검출이 가능하였고 전체 과정의 수행에 단지 5시간 정도 걸렸다. S. enteritidis 섬모항원 유전자를 cloning한 pGem-4-Sef B(-) DNA와 인위적으로 접종된 우유로부터 추출한 Salmonella DNA를 함께 중합효소반응으로 증폭한 후 제한효소로 잘라 전기영동을 행하여 band의 강도를 비교함으로써 Salmonella DNA copy수를 추정하는 것이 가능하였다.

  • PDF

Regular Exercise Training Increases the Number of Endothelial Progenitor Cells and Decreases Homocysteine Levels in Healthy Peripheral Blood

  • Choi, Jeong Kyu;Moon, Ki Myung;Jung, Seok Yun;Kim, Ji Yong;Choi, Sung Hyun;Kim, Da Yeon;Kang, Songhwa;Chu, Chong Woo;Kwon, Sang Mo
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제18권2호
    • /
    • pp.163-168
    • /
    • 2014
  • Endothelial progenitor cells (EPCs) are known to play an important role in the repair of damaged blood vessels. We used an endothelial progenitor cell colony-forming assay (EPC-CFA) to determine whether EPC numbers could be increased in healthy individuals through regular exercise training. The number of functional EPCs obtained from human peripheral blood-derived AC133 stem cells was measured after a 28-day regular exercise training program. The number of total endothelial progenitor cell colony-forming units (EPC-CFU) was significantly increased compared to that in the control group (p=0.02, n=5). In addition, we observed a significant decrease in homocysteine levels followed by an increase in the number of EPC-CFUs (p=0.04, n=5), indicating that the 28-day regular exercise training could increase the number of EPC colonies and decrease homocysteine levels. Moreover, an inverse correlation was observed between small-endothelial progenitor cell colony-forming units (small-EPC-CFUs) and plasma homocysteine levels in healthy men (r=-0.8125, p=0.047). We found that regular exercise training could increase the number of EPC-CFUs and decrease homocysteine levels, thus decreasing the cardiovascular disease risk in men.

H1975 세포에서 황금추출물에 의한 gefitinib 저항성 억제 효과 (Root Extract of Scutellaria Baicalensis Increases Gefitinib Sensitivity in H1975 Human Non-small Cell Lung Cancer Cells)

  • 박신형;박현지
    • 동의생리병리학회지
    • /
    • 제35권4호
    • /
    • pp.117-123
    • /
    • 2021
  • Gefitinib, a first generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR TKI), provides obvious clinical benefit in patients with EGFR-mutant non-small cell lung cancer (NSCLC). However, patients ultimately develop gefitinib resistance which mainly caused by EGFR T790M secondary mutation. In the current study, we investigated whether the root extract of Scutellaria baicalensis (SB) overcomes gefitinib resistance. Gefitinib-resistant H1975 human NSCLC cells (EGFR L858R/T790M double mutant) were treated with gefitinib and/or ethanol extract of SB (ESB) to evaluate the effect of ESB on the gefitinib sensitivity. The cell viability was measured by MTT assay and trypan blue exclusion assay. The colony-forming ability was evaluated by anchorage-dependent colony formation assay. Combined treatment with gefitinib and ESB markedly decreased the cell viability and colony formation than single treatment with gefitinib or ESB in H1975 cells. In addition, cells treated with both gefitinib and ESB exhibited a significant increase of sub-G1 DNA content which indicates apoptotic cells compared with those treated with gefitinib or ESB alone. As a molecular mechanism, combined treatment with gefitinib and ESB strongly downregulated the phosphorylation of ERK and JNK than single treatment with gefitinib or ESB. Taken together, our results demonstrate that ESB sensitizes H1975 cells to gefitinib treatment. We cautiously propose that ESB can be used in combination with gefitinib for the advanced NSCLC patients with acquired resistance to EGFR TKIs.

도두(刀豆)의 조혈증진 작용에 관한 연구 (Enhancing effect of Canavalia gladiata DC semen on the hematopoietic expansion and function of stem cells)

  • 김승형;김근회;지경엽;조인식;김한영;이영철
    • 대한본초학회지
    • /
    • 제27권4호
    • /
    • pp.9-16
    • /
    • 2012
  • Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

가미하고초산(加味夏枯草散)이 항암제(抗癌劑)의 항종양효과(抗腫瘍效果)와 종양세포(腫瘍細胞)에 미치는 영향(影響) (Influence of Gamihagochosan on The Antitumor effect of Anticancer Drug and The Proliferation of Tumor Cell Lines)

  • 김균택;전병훈
    • 대한한방내과학회지
    • /
    • 제18권1호
    • /
    • pp.175-190
    • /
    • 1997
  • In order to investigate the effects of Gamihagochosan Extract(加味夏枯草散抽出液) on antitumor effects after human cell lines (A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurement of antitumor effect of Mitomycin C(MMC) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for the measurement of antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows : 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 Cells after exposure to the extract of Gamihagochosan extract depressed the growth of tumor cells by concentration of Garnihagochosan. 2. Antitumor activity of the ethanol extract from Gamihggochosan extract and MMC on ascites form of Ehrlich carcinoma in mice is slightly improved. Especially the mean of survival times in the group of 200mg/kg and MMC 0.1mg/kg is improved over 34.9%. 3. When Gamihggochosan extract and MMC are administered together, the weight of tumor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Gamihagochosan extract and MMC are more significantly improved than MMC alone. According to the above result, it could be suggested that Gamihagochosan extract has indirect antitumor effect by the increase of MMC uptake.

  • PDF

죽엽석고탕가감방(竹葉石膏湯加減方)이 항암화학요법제(抗癌化學療法劑)의 세포독성(細胞毒性)과 종양세포(腫瘍細胞)의 성장억제(成長抑制)에 미치는 효과(效果)에 대한 연구 (Effects of Jukyeopseokgotanggagambang on Anti-tumor Chemotherapeutic Cytotoxicity and Lysosomal Enzymes of Tumor Cell)

  • 전승훈;문구;전병훈
    • 대한한방내과학회지
    • /
    • 제18권1호
    • /
    • pp.191-206
    • /
    • 1997
  • In order to investingate the effects of Jukyeopseokgotanggagambang Extract on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI 1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. According to the change of colony-forming efficiency and SRB assay of Caki-1 cell, hep3B and A549 cells after exposure to the extract of Jukyeopseokgotanggagambang extract, that extract depressed the growth of tumor cells depending on its concentration. 2. Antitumor activities of the ethanol extract from Jukyeopseokgotanggagambang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of Jukyeopseokgotanggagambang extract 200mg/kg and MMC 0.1mg/kg is improved over 30%. 3. When Jukyeopseokgotanggagambang extract and MMC are administerated together, the weight of tumor is more decreased than MMC alone. 4. The effects of the Jukyeopseokgotanggagambang extract and MMC on the lysosomal enzymes in Ehrlich ascites carcinoma cell are more significantly improved than MMC alone. 5. Jukyeopseokgotanggagambang extract also increased the uptake of MMC into Ehrlich carcinoma cells. According to the above results, it could be suggested that Jukyeopseokgotanggagambang extract has indirect autitumor effects by strengthening the effects of MMC on tumor cells.

  • PDF