• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.347-352
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• 1994

Cytotoxic test was performed by SRB assay on human epidermoid carcinoma HEp-2 cell line for screening the soil microorganism, secreting anticancer agent. One microorganism was selected among two thousand microorganisms for its highest cytotoxicity. And this microorganism was identified with Streptomyces species after performing of diaminopimeric acid and reducing sugar analysis of cell wall and analysing the cultural characteristics and named Streptomyces sp. SM 1119. The effect of anticancer agent in SM 1119 culture extract on the cell cycle was studied by using GG$_{o}$ synchronized NIH 3T3 cells. The extract inhibited the serum stimulation of GG$_{o}$ NIH 3T3 cell only within 1 hour after serum stimulation but not after 6 hours. The extract also reduced the amount of c-myc mRNA in Colo 320 cell. These results suggest that the anticancer agent in the extract inhibits the progression of cell cycle very early stages, probably from G$_{0}$ to G$_{1}$.

• Plant Resources
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• v.4 no.3
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• pp.181-188
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• 2001

We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.581-584
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• 2002

Individual surface(hydrophilic/hydrophobic) were prepared and mammalian cells were cultured on the hydrophilic region. For drug test, cancer and normal cells were treated with Taxol, as an example. Our system was compared with MTT assay. CHO cells were resistant to Taxol up to 100 nM in both Methods. However, A549 cells was sensitive at 100 nM Taxol in the 2 day-treatment. Cervical carcinoma cell, HeLa, was very sensitive to Taxol. In our system, the cells were not shown from above 20 nM Taxol treatment. Our system was competitive to MTT assay in animal cells for drug test.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.6909-6912
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• 2015

Cancer is a major public health problem due to the jeavy disease burden, fatality and tendency for increased incidence. Of all cancer types, cervical cancer is reported to be the fourth most common cancer in women, and the seventh overall, with an estimated 528,000 new cases worldwide in 2012. It features a long preclinical phase with slowly progressing precancerous lesions such as CIN 2 and 3 and adenocarcinoma in situ. Therefore, screening programs such as with Pap smear tests may play an important role in cervical cancer prevention. The purpose of this study was to present results of a Pap smear screening survey for cervical cancer targeting women living in an urban area in the province of Mersin, located in the Mediterranean region of Turkey. This community-based descriptive study included women living at Akdeniz county of Mersin province. A total of 1,032 screened women between 30 and 65 ages within the routine screening programme constituted the study population. The mean age of the participants was $43.8{\pm}8.6$ (min. 30, max. 65) years. The percentage of the participants who had previously undergone smears was 40.6%. Epithelial cell changes were found in 26 (2.5%) participants, with ASC-US in 18 (1.7%), ASC-H in 2 (0.2%), LSIL in 5 (0.5%) and HSIL in 1 (0.1%). The most common clinical presentation together with epithelial changes was abnormal vaginal discharge. Taking into account the presence of women who had never undergone Pap test; it should be offered at primary level of health care in the form of a community-based service to achieve reduced morbidity and mortality rates.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.975-981
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• 2014

Cervical cancer is an issue of foremost importance globally, specifically affecting the developing nations. Significant advances have taken place with regard to diagnosis of cervical cancer, especially with screening. Appropriate screening measures can thus reduce the incidence of cervical cancer. The most desirable screening technique should be less invasive, easy to perform, cost-effective and cover a wide range of diagnostic icons. Manual liquid based cytology (MLBC) can be considered as one of the suitable technique for screening with the above-mentioned benefits. The aim of the current study was to compare two cervical screening techniques on the basis of different morphological parameters and staining parameters by using modified acetic acid Pap staining to see the possibility of reducing time economy involved in conventional Pap staining (CPS). The study was conducted on a total 88 cases and all were analyzed with both MLBC and CPS. Forty eight cases that were regarded as satisfactory on the basis of Bethesda system by both methods were further recruited for investigation. Their morphological parameters and staining quality were compared and scored according to a scoring system defined in the study. Quality indices was calculated for both staining procedures and smear techniques.

• Journal of Nutrition and Health
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• v.35 no.10
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• pp.1053-1059
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• 2002

In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

• Molecules and Cells
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• v.42 no.6
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• pp.480-494
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• 2019

Aggregates of disease-causing proteins dysregulate cellular functions, thereby causing neuronal cell loss in diverse neurodegenerative diseases. Although many in vitro or in vivo studies of protein aggregate inhibitors have been performed, a therapeutic strategy to control aggregate toxicity has not been earnestly pursued, partly due to the limitations of available aggregate models. In this study, we established a tetracycline (Tet)-inducible nuclear aggregate (${\beta}23$) expression model to screen potential lead compounds inhibiting ${\beta}23$-induced toxicity. High-throughput screening identified several natural compounds as nuclear ${\beta}23$ inhibitors, including peucedanocoumarin III (PCIII). Interestingly, PCIII accelerates disaggregation and proteasomal clearance of both nuclear and cytosolic ${\beta}23$ aggregates and protects SH-SY5Y cells from toxicity induced by ${\beta}23$ expression. Of translational relevance, PCIII disassembled fibrils and enhanced clearance of cytosolic and nuclear protein aggregates in cellular models of huntingtin and ${\alpha}$-synuclein aggregation. Moreover, cellular toxicity was diminished with PCIII treatment for polyglutamine (PolyQ)-huntingtin expression and ${\alpha}$-synuclein expression in conjunction with 6-hydroxydopamine (6-OHDA) treatment. Importantly, PCIII not only inhibited ${\alpha}$-synuclein aggregation but also disaggregated preformed ${\alpha}$-synuclein fibrils in vitro. Taken together, our results suggest that a Tet-Off ${\beta}23$ cell model could serve as a robust platform for screening effective lead compounds inhibiting nuclear or cytosolic protein aggregates. Brain-permeable PCIII or its derivatives could be beneficial for eliminating established protein aggregates.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.2
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• pp.1200-1206
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• 2015

The major target for drug discovery, G-protein coupled receptor (GPCR) is involved in many physiological activities and related to various diseases and disorders. Among experimental techniques relating to the GPCR drug discovery process, various cell-based screening methods are influenced by cell conditions used in the overall process. Recently, the utilization of frozen cells is suggested in terms of reducing data variation and cost-effectiveness. The aim of this study is to evaluate various conditions in cell freezing such as temperature conditions and storage terms. The stable cell lines for calcium sensing receptor and urotensin receptor were established followed by storing cultured cells at $-80^{\circ}C$ up to 4 weeks. To compare with cell stored at liquid nitrogen, agonist and antagonist responses were recorded based on the luminescence detection by the calcium induced photoprotein activation. Cell signals were reduced as the storage period was increased without the changes in $EC_{50}$ and $IC_{50}$ values $EC_{50}:3.46{\pm}1.36mM$, $IC_{50}:0.49{\pm}0.15{\mu}M$). In case of cells stored in liquid nitrogen, cell responses were decreased comparing to those in live cells, however changes by storage periods and significant variations of $EC_{50}/IC_{50}$ values were not detected. The decrease of cell signals in various frozen cells may be due to the increase of cell damages. From these results, the best way for a long-term cryopreservation is the use of liquid nitrogen condition, and for the purpose of short-term storage within a month, $-80^{\circ}C$ storage condition can be possible to adopt. As a conclusion, the active implementation of frozen cells may contribute to decrease variations of experimental data during the initial cell-based screening process.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.64-64
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• 1993

당해년도에는 HBV-X와 결합하는 상대 단백질을 탐색하는 것이 주목적이다. 이 목적으로 1) 우선 생물학적 활성을 보유하는 X-단백질을 E. coil system에서 다량으로 생산하는 공정을 확립하였으며 2) 이 X-단백질을 labelling한 후 Probe로 사용하여 liver cell내에 존재하는 43Kd, 48Kd, 55Kd, 100Kd의 단백질이 HBV-X에 결합하는 것을 확인하였으며 3) liver cell expression library를 screening하여 HBV-X와 결합하는 단백질을 coding하는 유전자를 cloning하여 현재 각 clone들을 규명하고 있는 중이다. 4) 또한 암억제 유전자 산물인 p53과 X-단백질과의 상호작용을 발견하였다. 이러한 결과는 X-단백질이 간암의 발생에 작용하는 기작을 설명할 수 있는 중요한 발견이다.

• Molecules and Cells
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• v.44 no.3
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• pp.127-135
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• 2021

Since the introduction of RNA sequencing (RNA-seq) as a high-throughput mRNA expression analysis tool, this procedure has been increasingly implemented to identify cell-level transcriptome changes in a myriad of model systems. However, early methods processed cell samples in bulk, and therefore the unique transcriptomic patterns of individual cells would be lost due to data averaging. Nonetheless, the recent and continuous development of new single-cell RNA sequencing (scRNA-seq) toolkits has enabled researchers to compare transcriptomes at a single-cell resolution, thus facilitating the analysis of individual cellular features and a deeper understanding of cellular functions. Nonetheless, the rapid evolution of high throughput single-cell "omics" tools has created the need for effective hypothesis verification strategies. Particularly, this issue could be addressed by coupling cell engineering techniques with single-cell sequencing. This approach has been successfully employed to gain further insights into disease pathogenesis and the dynamics of differentiation trajectories. Therefore, this review will discuss the current status of cell engineering toolkits and their contributions to single-cell and genome-wide data collection and analyses.