• 한국식품영양과학회지
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• 제39권8호
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• pp.1107-1112
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• 2010

발아 전후 6종의 벼(일품벼, 백진주벼, 설갱벼, 고아미2, 거대배아벼 및 흑광벼)의 물 및 에탄올 추출물에 대한 암세포주 성장억제효과를 간암세포주(HepG2), 전립선암세포주(PC-3) 및 유방암세포주(MCF-7)에 대하여 살펴보았다. 물추출물은 1 mg/mL 농도에서 3종의 암세포주에 대하여 118.97~44.85% 범위의 높은 생존율을 보여 항암효과가 낮았지만 에탄올 추출물은 90.82~5.04% 범위의 생존율을 보여 물 추출물보다 높은 암세포 성장억제 효과를 나타내었다. 일품벼 발아 전후 에탄올 추출물은 간암세포주에 대하여 1 mg/mL 농도에서 각각 27.23 및 42.36%의 생존율을 보여 발아 시 항암효과가 증가하였으며, 특히 유방암세포주에 대해서는 0.25 mg/mL 농도에서 각각 48.20 및 56.82%의 생존율을 보여 다른 품종보다 성장억제율이 높았다. 또한 설갱벼발아 전후 에탄올 추출물은 전립선암세포에 대하여 1 mg/mL 농도에서 각각 44.52 및 64.51%의 생존율을 보여 발아 항암효과가 증가하였다. 추후 항암활성 물질의 분리 동정과 메커니즘 규명에 대한 연구가 수행되어야 할 것으로 판단된다.

• Animal cells and systems
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• 제8권3호
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• pp.213-220
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• 2004

MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5799-5804
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• 2012

Family with sequence similarity 189, member B (FAM189B), alias COTE1, a putative oncogene selected by microarray, for the first time was here found to be significantly up-regulated in hepatocellular carcinoma (HCC) specimens and HCC cell lines. mRNA expression of COTE1 in HCC samples and cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR, while protein expression of COTE1 in HCC tissues was assessed by immunohistochemistry. In addition, invasion of HCC cells was observed after overexpressing or silencing COTE1. In the total of 48 paired HCC specimens, compared with the adjacent non-cancer tissues, the expression of COTE1 was up-regulated in 31 (p<0.01). In HCC cell lines, COTE1 expression was significantly higher than in normal human adult liver (p<0.01). Overexpression of COTE1 enhanced HCC-derived LM6 and MHCC-L cellular invasion in vitro. In contrast, COTE1 knockdown via RNAi markedly suppressed these phenotypes, as documented in LM3 and MHCC-H HCC cells. Mechanistic analyses indicated that COTE1 could physically associate with WW domain oxidoreductase (WWOX), a tumor suppressor. COTE1 may be closely correlated with invasion of hepatocellular carcinoma (HCC) cells and thus may serve as an effective target for gene therapy.

• IMMUNE NETWORK
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• 제13권4호
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• pp.133-140
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• 2013

Since the discovery of the immunomodulation property of mesenchymal stem cells (MSCs) about a decade ago, it has been extensively investigated whether MSCs can be used for the treatment of immune-related diseases, such as graft versus-host disease (GvHD). However, how to evaluate the efficacy of human MSCs for the clinical trial is still unclear. We used an MHC-mismatched model of GvHD (B6 into BALB/c). Surprisingly, the administration of the human MSCs (hMSCs) could reduce the GvHD-related mortality of the mouse recipients and xenogeneically inhibit mouse T-cell proliferation and $IFN-{\gamma}$ production in vitro. We recently established a new protocol for the isolation of a homogeneous population of MSCs called subfractionation culturing methods (SCM), and established a library of clonal MSC lines. Therefore, we also investigated whether MSCs isolated by the conventional gradient centrifugation method (GCM) and SCM show different efficacy in vivo. Intriguingly, clonal hMSCs (hcMSCs) isolated by SCM showed better efficacy than hMSCs isolated by GCM. Based on these results, the MHC-mismatched model of GvHD may be useful for evaluating the efficacy of human MSCs before the clinical trial. The results of this study suggest that different MSC lines may show different efficacy in vivo and in vitro.

• Nutrition Research and Practice
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• 제4권3호
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• pp.177-182
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• 2010

The Chaga mushroom (Inonotus obliquus) has been used in folk medicine to treat cancers. However, limited information exists on the underlying anticancer effects of the major component of I. obliquus in vivo. We hypothesize that the pure compounds ($3{\beta}$-hydroxy-lanosta-8,24-dien-21-al, inotodiol and lanosterol, respectively) separated from I. obliquus would inhibit tumor growth in Balbc/c mice bearing Sarcoma-180 cells (S-180) in vivo and growth of human carcinoma cells in vitro. To test this hypothesis, the growth inhibition of each subfraction isolated from I. obliquus on human carcinoma cell lines (lung carcinoma A-549 cells, stomach adenocarcinoma AGS cells, breast adenocarcinoma MCF-7 cells, and cervical adenocarcinoma HeLa cells) was tested in vitro. Then, after S-180 implantation, the mice were fed a normal chow supplemented with 0, 0.1 or 0.2 mg of subfraction 1, 2 or 3 per mouse per day. All of the subfractions isolated from I. obliquus showed significant cytotoxic activity against the selected cancer cell lines in vitro. Subfraction 1 was more active than subfraction 2 and subfraction 3 against the A549, AGS and MCF-7 cancer cell lines in vitro. In in vivo results, subfraction 1 isolated from I. obliquus at concentrations of 0.1 and 0.2 mg/mouse per day significantly decreased tumor volume by 23.96% and 33.71%, respectively, as compared with the control. Subfractions 2 and 3 also significantly inhibited tumor growth in mice bearing S-180 as compared with the control mouse tumor. Subfraction 1 isolated from I. obliquus showed greater inhibition of tumor growth than subfractions 2 and 3, which agrees well with the in vitro results. The results suggest that I. obliquus and its compounds in these subfractions isolated from I. obliquus could be used as natural anticancer ingredients in the food and/or pharmaceutical industry.

• BMB Reports
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• 제40권3호
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• pp.358-367
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• 2007

A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 $\mu$g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 $\pm$ 6.8), Bre-04 (41.5 $\pm$ 4.8), and Lu-04 (11.4 $\pm$ 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with $TC_50$ and $EC_50$ of 5.176 mg/ml and 3.054 $\mu$g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9445-9451
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• 2014

Neuroblastoma is the most common extracranial solid tumor in children. Approximately half of the affected patients are diagnosed with high-risk poor prognosis disease, and novel therapies are needed. Sanguinarine is a benzophenanthridine alkaloid which has anti-microbial, anti-oxidant and anti-inflammatory properties. The aim of this study is whether sanguinarine has in vitro apoptotic effects and which apoptotic genes might be affected in the human neuroblastoma cell lines SH-SY5Y (N-myc negative), Kelly (N-myc positive, ALK positive), and SK-N-BE(2). Cell viability was analysed with WST-1 and apoptotic cell death rates were determined using TUNEL. After RNA isolation and cDNA conversion, expression of 84 custom array genes of apoptosis was determined. Sanguinarine caused cell death in a dose dependent manner in all neuroblastoma cell lines except SK-N-BE(2) with rates of 18% in SH-SY5Y and 21% in Kelly human neuroblastoma cells. Cisplatin caused similar apoptotic cell death rates of 16% in SH-SY5Y and 23% in Kelly cells and sanguinarine-cisplatin combinations caused the same rates (18% and 20%). Sanguinarine treatment did not affect apoptototic gene expression but decreased levels of anti-apoptotic genes NOL3 and BCL2L2 in SH-SY5Y cells. Caspase and TNF related gene expression was affected by the sanguinarine-cisplatin combination in SH-SY5Y cells. The expression of regulation of apoptotic genes were increased with sanguinarine treatment in Kelly cells. From these results, we conclude that sanguinarine is a candidate agent against neuroblastoma.

• International Journal of Oral Biology
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• 제42권3호
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• pp.107-115
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• 2017

Human malignant melanoma is an aggressive skin cancer which has been rising at a greater rate than any other cancers. Although various new therapeutic methods have been developed in previous studies, this disease has properties of high proliferation and metastasis rate which remain obstacles that have lead to a poor prognosis in patients. It has been reported that a specific Lactobacillus extract has anti-cancer and -metastasis effect in vitro and in vivo. However, previous research has not specified precisely what effect the Lactobacillus rhamnosus GG (LGG) extract has had on human malignant melanomas. In this study, we showed that the LGG extract has anti-cancer and -metastasis effects on the human malignant melanoma cell lines, A375P and A375SM. At first, it was found that, while the LGG extract affects human neonatal dermal fibroblasts slightly, it induced the dose-dependent anti-cancer effect on A375P and A375SM by a WST-1 proliferation assay. As a result of a real-time PCR analysis, the expression patterns of several genes related to cell cycle, proliferation, and apoptosis were modulating in a manner that inhibited the growth of both malignant melanoma cell lines after the treatment of the LGG extract. Furthermore, genes related to the epithelial-mesenchymal transition were down-regulated, and migration rates were also decreased significantly by the LGG extract. Our study showed that the LGG extract could be used as a potential therapeutic source.

• Tuberculosis and Respiratory Diseases
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• 제42권3호
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• pp.302-313
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• 1995

연구배경: 종양괴사인자(Tumor necrosis factor; TNF)는 다양한 생물학적인 작용을 가지며 종양 세포에 대한 세포 독성은 그 대표적인 기능중의 하나이다. TNF-$\alpha$는 생체외에서(in vitro) 몇몇 종양 세포주에 대하여 항암제, 특히 topoisomerase II targeted chemotherapeutic agent의 세포 독성 효과를 상승적으로 증가시키는 것이 알려져 있다. 최근 암세포에 대한 cytokine 유전자 요법에서 TNF는 중요한 대상으로 여겨지고 있으며, 유전자 이입에 의해 암조직이 TNF를 생성하게 될 경우 암 증식 억제 효과가 있음이 보고되고 있다. 연구자는 암세포에 TNF-$\alpha$ 유전자를 이입하여 자신이 TNF-$\alpha$를 생성하도록 형질을 변환시킨 암세포는 topoisomerase II 억제 항암제에 대한 김수성에 변화가 있을 것이라는 가설을 수립하였고 이를 검증하고자 본 연구를 수행하였다. 본 연구에서는 생체외로(in vitro) TNF-$\alpha$ 유전자를 이입하여 TNF-$\alpha$를 생성하는 암세포주에서 topoisomerase II targeted drug에 대한 항암제 감수성 효과가 모세포주에 비하여 증대될 수 있는지를 알아 보고자하였다. 방법: TNF-$\alpha$에 감수성을 보이는 것으로 알려진 인체 중피종 세포주인 NCI-H2058 세포주 및 생쥐의 섬유육종 세포주인 WEHI164 세포주와 인체 비소세포 폐암 세포주인 A549 세포주를 배양하여, 먼저 임상에서 흔히 폐암의 항암 화학 요법 치료에 널리 쓰이는 대표적인 topoisomerase II targeted chemotherapeutic drug인 etoposide(VP-16)와 doxorubicin(adriamycin)을 가하였을 때 관찰된 세포 독성을 MTT assay로 측정하고, 각 모세포주(parenta1 cell line)에 TNF-$\alpha$의 유전자를 이입시켜서 형절 변환한 세포주(transformed cell line)에 대하여 각각 동일한 항암제를 가하였을 때 관찰된 세포 독성의 정도를 같은 방법으로 측정하여, 그 결과를 비교 분석하였다. 또한 모세포주에 외부에서 TNF를 가하여 전처치한 후 동일한 항암제를 가하였을 때의 세포독성을 관찰하여 비교 분석하였다. 결과: H2058 세포주에서는 TNF-$\alpha$ 유전자를 이입한 세포주 topoisomerase II targeted drug을 가하였을 때, 항암제 감수성이 모세포주에 같은 항암제를 가하였을 때에 비하여 의미있게 증가함을 관찰할 수 있었으나(p<0.05), WEHI 세포주와 A549 세포주에 있어서는 TNF-$\alpha$ 유전자를 이입한 세포주에서 모세포주에 비하여 항암제 감수성이 증가하지는 않았다. 결론: TNF-$\alpha$ 유전자의 이입이 topoisomerase II targeted chemotherapeutic drug에 대한 항암제 감수성을 증가시키는 효과는 세포주에 따라 다양한 결과를 보이는 것을 알 수 있었으며, 적어도 선택된 특정 종류의 호흡기계 암세포에 있어서는 TNF-$\alpha$ 유전자의 이입으로 항암제 감수성(chemosensitivity)을 증가시킬 수 있을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.810-815
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• 2004

Many biological properties and the clinical potential of human interleukin-2 (hIL-2) draw much attention to its high-level expression in mammalian cells. Recombinant human IL-2 (rhIL-2) was expressed in Chinese hamster ovary (CHO) cells, using the recently developed expression system which confers position-independent expression. Stable CHO cell lines carrying several hundred amplified copies of the rhIL-2 gene were easily obtained and rhIL-2 was expressed at high levels after selection with increasing concentrations of methotrexate. Interestingly, the insertion of the transcription terminator of the human gastrin gene into the downstream region of the gene for rhIL-2 considerably increased rhIL-2 expression. Using the expression system with the transcription terminator, it was possible to get a CHO cell line expressing the rhIL-2 at a very high level, about $11.4\mug/10^6$ cell/day, which is about 6 times higher than that previously reported. The biological activity of the rhIL-2 protein purified from the cell line was also confirmed by the cell proliferation assay.