• Food Science and Biotechnology
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• 제16권1호
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• pp.163-166
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• 2007

The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

• Molecules and Cells
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• 제26권5호
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• pp.514-516
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• 2008

The cancer stem cell hypothesis posits that tumor growth is driven by a rare subpopulation of cells, designated cancer stem cells (CSC). Studies supporting this theory are based in large part on xenotransplantation experiments wherein human cancer cells are grown in immunocompromised mice and only CSC, often constituting less than 1% of the malignancy, generate tumors. Herein, we show that all colonies derived from randomly chosen single cells in mouse lung and breast cancer cell lines form tumors following allografting histocompatible mice. Our study suggests that the majority of malignant cells rather than CSC can sustain tumors and that the cancer stem cell theory must be reevaluated.

• 대한한방종양학회지
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• 제2권1호
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• pp.43-56
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• 1996

The sprig of Sojekbojungwhan(消積保中丸) has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Sojekbojungwhan extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell lines(A549, hep3B, Caki-1, Sarcoma 180) after esposure to Sojekbojungwhan extract using in ILS, colony forming efficiency 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. As a result of exposure to Sojekbojungwhan extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Sojekbojungwhan extact showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Sojekbojungwhan decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Sojekbojungwhan also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Sojekbojungwhan extract into TBM. 4. Sojekbojungwhan extract also increased NK cell activities. According to the above results, it could be suggested that Sojekbojungwhan extract has some antitumor effects.

• IMMUNE NETWORK
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• 제5권2호
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• pp.124-129
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• 2005

Background: CM1 (centrocyte/-blast marker 1) is originally defined as a germinal center B cell marker. It is known that CM1 plays a critical role on B cell development in germinal center. In addition, we have found that CM1 is expressed on lymphoma cell lines, such as Raji, Ramos and IM-9. This means that CM1 might be served as a tumor marker as well. In the present study, we examined the expression of CM1 on the surface of the other tumors and the possibility of the development of tumor screening ELISA kit by using CM1. Methods: First, we have examined the expression of CM1 on stomach cancer and hepatoma, which are predominantly (discovered) occurred in Korean, by flow cytometry analysis. After purifying of CM1 antigen from Raji and Ramos, the optimal ELISA condition was determined. And then we compared the level of CM1 between normal individuals and cancer patients by ELISA. To decrease the non-specific binding of anti-CM1 mAb with serum components except CM1 and to enhance the diagnostic accuracy, albumin depletion spin column was used. Results: CM1 was highly expressed on stomach cancer and hepatoma cell lines. In addition, we have also confirmed the increased CM1 expression on cancer patients. The difference of CM1 expression between normal individuals and cancer patients were more clearly observed, after deletion of serum albumin by using albumin depletion spin column. Conclusion: Based on the results from this study, CM1 might be a useful molecule for the early diagnosis of cancer. In addition, further studies for the increase of ELISA sensitivity and appropriate albumin depletion methods should be needed.

• 대한약침학회지
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• 제23권4호
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• pp.212-219
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• 2020

Objectives: Nowadays cancer treatment is an important challenge in the medical world that needs better therapies. Many active secretions produced by insects such as honey bees used to discover new anticancer drugs. Bee venom (BV) has a potent anti inflammatory, anti cancer and tumor effects. The aim of present study is evaluation of anticancer effects induced by Apis mellifera venom (AmV) on cell Lines. Methods: AmV was selected for study on cancer cell lines. Total protein, molecular weight and LD50 of crude venom were determined. Then, cells were grown in Dulbecco's Modified Eagle medium supplemented with 10% fetal bovine serum and 1% antibiotics. The A549, HeLa and MDA-MB-231 cell Lines were exposed by different concentration of AmV. The morphology of cells was determined and cell viability was studed by MTT assay. Evaluation of cell death was determined by and DNA fragmentation. Results: The results from MTT assay showed that 3.125 ㎍/mL of A549, 12.5 for HeLa and 6.25 ㎍/mL of MDA-MB-231 killed 50% of cells (p < 0.05). Morphological analysis and the results from hoescht staining and DNA fragmentation indicated that cell death induced by AmV was significantly apoptosis. Conclusion: The data showed that using lower dosage of AmV during treatment period cause inhibition of proliferation in time and dose dependant manner. Findings indicated that some ingredients of AmV have anticancer effects and with further investigation it can be used in production of anticancer drugs.

• Tuberculosis and Respiratory Diseases
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• 제41권2호
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• pp.87-96
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• 1994

연구배경 : 종양괴사인자(tumor necrosis factor ; TNF)는 여러 암세포에 대해서 세포독성을 보임이 알려져 있다. 그리고, 최근 분자생물학적 발전에 힘입어 TNF 유전자를 암세포에 이입하고 발현시키는 연구가 그 동안 진행되었다. 이들 연구의 목적은, TNF를 전신적으로 쓸 경우 전신 부작용이 심하여 인체에 쓸 수 없는 현 단계에서 암세포 자체에서 TNF를 만들어 내어서 암세포 주위에서만 작용하게 함으로써 연체에 미치는 전신독성을 최소한으로 줄이고 암세포를 사멸시키는 것이었다. 암세포에 TNF 유전자를 이입함으로써 예상되는 항암효과가 성공을 거두기 위해서는 첫째, TNF 유전자가 이입된 암세포에서 분비된 TNF가 주위 암세포를 성공적으로 사멸시켜야 하고 둘째는 분비된 TNF가, TNF 유전자가 이입된 암세포 자신을 사멸시켜야 한다. 본 연구는 이 중 두번째 기전, 즉 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 의해서 사멸되거나 또는 세포 독성이 나타날 수 있는가를 검증하는데 목적을 두었다. 방법 : TNF에 감수성을 보이는 인체의 중피종 세포주인 NCI-H2058과 생쥐 섬유육종 세포주인 WEHI164에 TNF-$\alpha$ 유전자를 retroviral vector를 이용하여 이입하고 TNF를 발현을 시도하였다. DNA 수준과 단백질 수준에서 TNF-$\alpha$ 유전자가 제대로 이입되어 발현되는지 PCR과 ELISA 및 bioassay(MTT assay)로 확인하였다. 그리고, TNF 유전자가 이입된 세포주가 자신이 분비하는 TNF에 사멸되는지 아니면 생존하는지 MTT(dimethylthiazolyl diphenyltetrazolium) assay로 알아보았다. 그리고, 만일 TNF 유전자가 이입된 암세포가 자신이 분비한 TNF에 사멸되지 않고 생존할 경우, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 암세포주가 외부에서 준 TNF에도 내성을 보이는지도 추가로 MTT assay 방법으로 확인해 보았다. 결과 : 1) TNF-$\alpha$ 유전자 이입 및 발현 확인 PCR을 시행한 결과, TNF 유전자가 이입된 NCI-H2058-TNF와 WEHI164-TNF 세포주는 790 base pair 크기의 진한 DNA band를 보인 반면 각각의 모세포주에서는 보이지 않아서 retroviral vector를 이용한 유전자 이입이 DNA 수준에서 이루어졌음을 확인할 수 있었다. 그리고, NCI-H2058-TNF와 WEHI164-TNF의 상층 배양액의 TNF양을 ELISA와 bioassay(MTT assay)로 측정한 결과, 생물학적 활성을 지닌 TNF를 각각 $23.6{\pm}0.84ng$/24h/$10^6cells$, $12.2{\pm}0.36ng$/24h/$10^6cells$ 생산함을 알 수 있었다. 2) TNF 유전자 이입 전후, 암세포의 TNF에 대한 감수성 비교 TNF 유전자 이입 전후의 TNF에 대한 세포주의 감수성(세포사망율)을 TNF의 농도 변화에 따라 비교한 결과, NCI-H2058의 경우 TNF 농도 100ng/ml에서 모세포는 $25{\pm}3%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 그리고, WEHI-164의 경우도 TNF 농도 100ng/ml에서 모세포는 $73{\pm}5%$의 세포독성을 보인 반면 TNF 유전자 이입 후에는 $3{\pm}2%$의 세포독성을 보여 통계적으로 유의한 차이가 있었다(p<0.01). 결론 : TNF에 감수성을 보이는 암세포주인 NCI-H2058과 WEHI164에 TNF 유전자 이입을 시행하고 TNF가 발현되게 하였을 때, TNF 유전자를 이입받은 두 암세포주 모두에서 자신이 생산해 내는 TNF에 내성을 보여 생존하였다. 뿐만 아니라 생존한 이들 세포는 외부에서 준 TNF에 대해서도 내성을 보였다. 따라서, 암세포에 대한 TNF 유전자 이입을 통한 유전자 요법이 성공을 거두려면 유전자 이입된 세포에서 분비하는 TNF의 면역 세포 동원 방법 등의 간접적인 항암기전이 필요할 것으로 생각된다.

• 대한암한의학회지
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• 제20권2호
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• pp.5-11
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• 2015

Objective : The objective of this study was to investigate the anti-tumor activity of the complexed herbal formula, Haeamdan (HAD). Methods : Seven Cancer cell lines, LoVo, MCF-7, AGS, Sarcoma 180, HL-60, NCI-H69, LL/2, were prepared and the cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2yl)-2,5-dephenyl tetrazolium bromide (MTT) assay. HAD was applied with various concentrations from 0.1 to 1.0 mg/ml to figure out the appropriate dosage. ICR male mice were intraperitoneally implanted with Sarcoma 180 and divided into 8 species for each group. Control group was treated with normal saline, positive control group was treated with cyclophosphamide 8mg/kg, and experimental group was treated with HAD 1g/kg. Results : Among seven cancer cell lines, HAD exhibited strong cytotoxic activities to followed four cancer cell lines, that is, Sarcoma 180, HL-60, NCI-H69, and LL/2. These cytotoxic activity was expressed under 0.50 mg/ml of IC50 under 0.1~1mg/ml of OBW. When Sarcoma 180 cancer cell was implanted in ICR male mice and treated with the HAD, HAD prolonged the median overall survival for 3.6 days, from 17.5 to 21.1 days. Conclusion : HAD showed strong cytotoxicity to the cancer cells, Sarcoma 180, HL-60, NCI-H69, on in vitro study and it showed anti-tumor activity in vivo with the peritoneal cancer mice by prolonging the median survival for 3.6 days. Further researches would be expected to support the anti-tumor efficacy of HAD.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2975-2979
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• 2015

Background: Telomere length is closely associated with cellular radiosensitivity and WRAP53 is required for telomere addition by telomerase. In this research we assessed radiosensitivity of laryngeal squamous cell carcinoma Hep-2 cell lines after WRAP53 inhibition, and analyzed the molecular mechanisms. Materials and Methods: phWRAP53-siRNA and pNeg-siRNA were constructed and transfected into Hep-2 cells with lipofectamine. Expression of WRAP53 was analyzed by RT-PCR and Western-blottin, radiosensitivity of Hep-2 cells was assessed colony formation assay, and the relative length of telomeres was measured by QPCR. Results: The data revealed that the plasmid of phWRAP53-siRNA was constructed successfully, and the mRNA and protein levels of WRAP53 were both obviously reduced in the Hep-2 cell line transfected with phWRAP53-siRNA. After Hep-2 cells were irradiated with X-rays, the $D_0$ and $SF_2$ were 2.481 and 0.472, respectively, in the phWRAP53-siRNA group, much lower than in the control group ($D_0$ and $SF_2$ of 3.213 and 0.592) (P<0.01). The relative telomere length in the phWRAP53-siRNA group was $0.185{\pm}0.01$, much lower than in the untreated group ($0.523{\pm}0.06$) and the control group ($0.435{\pm}0.01$). Conclusions: Decreasing the expression of WRAP53 using RNA interference technique can enhance the radiosensitivity of Hep-2 cell lines by influencing the telomere length. WRAP53 is expected to be a new target to regulate the radiosensitization of tumor cells.

• 한국축산식품학회지
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• 제24권3호
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• pp.293-297
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• 2004

본 연구에서는 유황오리로부터 항종양 효과를 나타내는 물질을 용매 추출법과 각종 크로마토그래피를 사용하여 분리 및 정제하였다. 유황오리로부터 정제한 활성 물질의 각종 종양세포에 대한 항암 효과를 측정한 결과 MTT assay는 100 $\mu\textrm{g}$/mL의 농도에서 HEp-2는 56%, KB는 58%의 세포 증식 억제 효과가 나타났다. 정상 세포주인 MDBK는 28%의 세포증식억제 효과가 나타나 정상 세포에 대한 세포 독성은 나타나지 않았다. Clonogenic assay는 200 $\mu\textrm{g}$/mL의 농도에서 HEp-2는 26%, KB는 28%의 생존율을 나타내었다. 따라서 유황오리로부터 정제한 활성 물질은 후두암 세포주인 HEp-2와 구강암 세포주인 KB에서만 특이적으로 세포 증식 억제 효과를 나타내었다.

• Bulletin of the Korean Chemical Society
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• 제25권12호
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• pp.1898-1906
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• 2004

Nonclassical quinazolinone analogs I, II, and III, in which the glutamic acid moiety of the classical antifolates is substituted by phenylglycine, phenylalanine or aminobenzoic acid and their methyl esters, were synthesized and evaluated as lipophilic inhibitors of thymidylate synthase (TS). The target compounds were generally potent inhibitors of L. casei and human TS with $IC_{50}$ values within the narrow range of 0.2-10 ${\mu}$M and 0.003-0.03 ${\mu}$M, respectively. Further, most of the target compounds showed cytotoxicity against tumor cell lines of murine and human origin with $IC_{50}$ values of as low as 0.050 ${\mu}$M. Substitution of another hydroxyl or carboxylic acid/ester group at the phenyl ring further increased the potency of TS inhibition and cell growth inhibition. Most effective were compounds If and Ic in which extra carboxylic acid/ester was present at the phenyl ring with nanomolar $IC_{50}$ values of 0.0044 and 0.0093 ${\mu}$M against human TS and submicromolar cytotoxic growth inhibition against all four tumor cell lines.