• BMB Reports
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• v.51 no.11
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• pp.602-607
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• 2018

Aberrant expression of microRNAs (miRNAs) plays important roles in carcinogenesis and tumor progression. However, the expression and biological role of miR-301b in triple-negative breast cancer (TNBC) remains unclear. Here we aimed to evaluate the roles and mechanisms of miR-301b in TNBC cells. miR-301b expression was assessed in TNBC specimens and cell lines by quantitative Real-Time PCR (qRT-PCR). TNBC cells were transfected with miR-301b mimics, inhibitors or Cylindromatosis (CYLD) small interfering RNA (siRNA) using Lipofectamine 2000. The functional roles of miR-301b were determined by cell proliferation, colony formation, and apoptosis assays. Western blots and qRT-PCR were used to measure the expression of mRNAs and proteins in the cells. We found that miR-301b was upregulated in TNBC specimens and cell lines. Overexpression of miR-301b promoted cell proliferation in TNBC cells, while inhibited the apoptosis induced by 5-FU. CYLD was downregulated by miR-301b at both mRNA and protein levels in TNBC cells. Dual-luciferase report assay confirmed that miR-301b downregulated CYLD by direct interaction with the 3'-untranslated region(3'-UTR) of CYLD mRNA. $NF-{\kappa}B$ activation was mechanistically associated with miR-301b-mediated downregulation of CYLD. However, inhibition of miR-301b reversed all the effects of miR-301b. In conclusion, miR-301b plays an oncogenic role in TNBC possibly by downregulating CYLD and subsequently activating $NF-{\kappa}B$ p65, and this may provide a novel therapeutic approach for TNBC.

• The Korean Journal of Food And Nutrition
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• v.23 no.4
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• pp.526-530
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• 2010

Isopentenyl diphosphate(IPP) isomerization to dimethylallyl diphosphate(DMAPP) is an important step for the efficient production of isoprenoids such as lycopene, ${\beta}$-carotene, astaxanthin, etc. The type II isopentenyl diphosphate isomerase gene from Synechocystis sp. PCC6803(sll1556, Syidi2) was cloned and expressed in Escherichia coli $DH5{\alpha}$. When E. coli $DH5{\alpha}$ harboring lycopene synthesis genes, crtE, crtB, and crtI and mevalonate pathway genes, MvK1, MvK2, and Mvd, was cultured on LB medium containing mevalonate, the strain grew very slowly be due to the toxicity of isopentenyl diphosphate derived from mevalonate. When Syidi2 was introduced to E. coli $DH5{\alpha}$ harboring the lycopene synthesis genes and mevalonate pathway genes, growth on mevalonate medium was fully restored and the colony showed red color indicating lycopene formation. The growth rate of the mutant strain, E. coli $DH5{\alpha}$(idi::${\Delta}km$), was very slow because of IPP accumulation and DMAPP deprivation. Ultimately the idi mutant was complemented by introducing the Syidi2 gene.

• Journal of Species Research
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• v.6 no.2
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• pp.101-118
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• 2017

In an investigation of indigenous prokaryotic species in Korea, a total of 57 bacterial strains assigned to the classes Betaproteobacteria and Gammaproteobacteria were isolated from diverse environments. Samples were collected from fresh water, natural caves, soil, paddy fields, lakes, sea water, jeotgal (fermented seafood), salt flats, soil from abandoned mines, plant roots, digestive organs of both Japanese crested ibis (Nipponia nippon) and Burmese python (Python molurus bivittatus) and tidal flats. From the high 16S rRNA gene sequence similarity (>98.7%) and formation of robust phylogenetic clades with closely related species, it was determined that each strain belonged to an independent and predefined bacterial species within either the Betaproteobacteria or Gammaproteobacteria. There is no official report or publication that describes these 57 proteobacterial species in Korea. Overall, in the class Betaproteobacteria there were 16 species in 12 genera of 4 families in the order Burkholderiales and two species in two genera of one family in the order Neisseriales. Within the class Gammaproteobacteia, there were five species in four genera of four families in the order Alteromonadales, 12 species in 11 genera of one family in the order Enterobacteriales, four species in four genera of three families in the order Oceanospirillales, 11 species in four genera of two families in the order Pseudomonadales, two species in the order Vibrionales and five species in five genera of one family in the order Xanthomonadales. Gram reaction, colony and cell morphology, basic biochemical characteristics, isolation source and strain IDs are described in the species description section.

• Korean Journal of Microbiology
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• v.39 no.2
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• pp.76-82
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• 2003

To prepare evolved PHB depolymerase with increased activity for PHB or P(3HB-co-3HV) compared to the activity of the original PHB depolymerase from Alcaligenes faecalis T1, random mutation of the cloned PHB depolymerase gene was performed by using a DNA shuffling method. A library of mutated PHB depolymerase genes from A. faecalis T1 was fused to the ice nucleation protein (INP) gene from Pseudomonas syringae in pJHCl 1 and approximately 7,000 transformants were isolated. Using M9 minimal medium containing PHB or P(3HB-co-3HV) as the carbon source, mutants showing alteration in PHB depolymerase activity were selected from the transformants. The PHB depolymease activity of the transformants was confirmed by the formation of halo around colony and the turbidity decrease tests using culture supermatants. The catalytic activity of PHB depolymerase of the best mutant II-4 for PHB or P(3HB-co-13 mol% 3HV) was approximately 1.8-fold and 3.2-fold, respectively, higher than that of the original PHB depolymerase. DNA sequence analysis revealed that three amino acid residues (Ala209Val, Leu258Phe, and Asp263Thr) were substituted in II-4. From the mutational analysis, it was presumed that the substitution of amino acids near catalytic triad to more hydrophobic amino acids enhance the catalytic activity of PHB depolymerase from A. faecalis T1.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.7
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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.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.2
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• pp.627-633
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• 2015

Background: To study the effect of parecoxib, a novel cyclooxygenase-2 selective inhibitor, on the radiation response of colorectal cancer (CRC) cells and its underlying mechanisms. Materials and Methods: Both in vitro colony formation and apoptosis assays as well as in vivo mouse xenograft experiments were used to explore the radiosensitizing effects of parecoxib in human HCT116 and HT29 CRC cells. Results: Parecoxib sensitized CRC cells to radiation in vitro with a sensitivity enhancement ratio of 1.32 for HCT116 cells and 1.15 for HT29 cells at a surviving fraction of 0.37. This effect was partially attributable to enhanced apoptosis induction by parecoxib combined with radiation, as illustrated using an in vitro apoptosis assays. Parecoxib augmented the tumor response of HCT116 xenografts to radiation, achieving growth delay more than 20 days and an enhancement factor of 1.53. In accordance with the in vitro results, parecoxib combined with radiation resulted in less proliferation and more apoptosis in tumors than radiation alone. Radiation monotherapy decreased microvessel density (MVD) and microvessel intensity (MVI), but increased the hypoxia level in xenografts. Parecoxib did not affect MVD, but it increased MVI and attenuated hypoxia. Conclusions: Parecoxib can effectively enhance radiation sensitivity in CRC cells through direct effects on tumor cells and indirect effects on tumor vasculature.

• Korean journal of applied entomology
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• v.55 no.3
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• pp.235-243
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• 2016

The winter cherry bug, Acanthocoris sordidus (Thunberg), is an insect pest hat damages plants from Solanaceae and Convolvulaceae. The developmental period from egg to adult averages 76 days at $25^{\circ}C$. Adult egg-laying occurred irregularly via spawning, averaging 195 (up to 468) eggs per individual on the abaxial leaf surface of the host plant. Results of linear regression indicated that the lower developmental threshold temperature was $13.9^{\circ}C$ and the effective accumulated temperature was 526.3 DD. Data from a pepper field in 2015 indicated that overwintering adults first appeared during late June (daily average temperature = $25.7^{\circ}C$), reaching maximum density by early September. A choice test examining colonization preferences using a net cage and a Y-tube olfactometer revealed that females gravitated toward conspecifics (other females, males, or both), whereas males moved toward empty areas. Finally, we found that communal breeding results in a longer developmental period and higher eclosion rates than solitary breeding. Developmental periods and eclosion rates were also for colonies in a large space than for those in a small space. This outcome suggests that colonization effects on insect development are stronger in a smaller area.

• Journal of the korean academy of Pediatric Dentistry
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• v.43 no.3
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• pp.306-312
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• 2016

Gargle solution has typically been used for the prevention of oral infectious disease such as dental caries and periodontitis. However, the use of most gargle solutions is controversial in application for children because some gargle solutions have harmful side effects. Electrolyzed water is generated by passed an electric current and has antimicrobial activity. The purpose of this study was to investigate and compare the efficacy of electrolyzed water in various conditions for eliminating cariogenic bacteria. Electrolyzed water was generated by a platinum electrode in the presence of sodium chloride at various concentrations. Streptococcus mutans and Streptococcus sobrinus were cultivated into a brain heart infusion broth. After harvesting planktonic bacteria, the pellets were treated with the electrolyzed water and commercial gargle solutions and plated on a mitis-salivarius agar plate. Also, the anti-biofilm activity of the electrolyzed water and commercial gargle solutions was investigated after biofilm formation of S. mutans and S. sobrinus. The bacteria in the biofilm were plated onto a mitis-salivarius agar plate. The plates were incubated, and the colony forming unit was measured. The electrolyzed water containing sodium chloride showed significant antibacterial activity against S. mutans and S. sobrinus as well as some gargle solutions. Furthermore, the electrolyzed water had more disruptive effect on the biofilm of S. mutans and S. sobrinus and killed more bacteria in the biofilm than commercial gargle solutions. The results demonstrate that electrolyzed water may be a useful gargle solution for prevention of dental caries.

• Korean Journal of Food Science and Technology
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• v.38 no.3
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• pp.419-426
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• 2006

A thermophilic microorganism, strain JE 375, which produces a thermostable protease, was isolated from soil and compost in Korea. This gram-positive, rod-shaped, catalase positive, motility positive, and hemolysis ${\beta}$ containing organism was implicated in glucose fermentation, mannitol fermentation, xylose oxidation, aerobic activity and spore formation. The color of the colony was yellowish white. The temperature range for growth at pH 6.5 was between 55 and $70^{\circ}C$, with an optimum growth temperature of $65^{\circ}C$. This result confirmed the strain JE 375 as a thermophilic microorganism. The enzyme was produced aerobically at $65^{\circ}C$ during 20 hr in a medium (pH 6.5) containing 1% trypton. 1% maltose, 0.5% yeast extract and 1% NaCl. The 16S rDNA of strain JE 375 had 97.6% sequence similarity with the 16S rDNA of Bacillus caldoxyloyticus. On the basis of biochemical and physiological properties and phylogenetic analysis, we named the isolated strain as Bacillus sp. JE 375. The thermostable protease from Bacillus sp. JE 375 had been partially purified and characterized. The molecular weight of the enzyme was deduced from SDS-PAGE and gel chromatography as 55 kDa and its optimal temperature was $60^{\circ}C$. The enzyme showed its highest activity at pH 7.5 and was stable from pH 7.0 to 8.0.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.17
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• pp.7297-7301
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• 2014

Objective: To explore the radiosensitization effect of overexpression of silent information regulator 6 (SIRT6) on A549 non-small cell lung cancer (NSCLC) cells. Methods: Adenovirus vector Ad-SIRT6 causing overexpression of SIRT6 was established. Western blotting and MTT assay were adopted to detect the level of SIRT6 protein and the inhibitory rate of A549 cell proliferation after different concentrations of adenovirus transduction (0, 25, 100, 200, and 400 pfu/cell) for 24 h. Control group, Ad-null group and Ad-SIRT6 group were designed in this experiment and virus concentration of the latter two groups was 200 pfu/cell. Colony formation assays were employed to test survival fraction (SF) of the 3 groups after 0, 2, 4, 6, 8, 10 X-ray irradiation. Flow cytometry was used to detect the status of cell cycle of 3 groups after 48 h of 4Gy X-ray irradiation and Western blotting was used to determine the expression of apoptosis-related genes of 3 groups after 48 h of 4GyX-ray irradiation. Results: In the range of 25~400 pfu/cell, the inhibitory rate of A549 cell proliferation increased as adenovirus concentration raised. The inhibitory rates under the concentrations of 0, 25, 100, 200, and 400 pfu/cell were 0%, $4.23{\pm}0.34%$, $12.7{\pm}2.57%$, $22.6{\pm}3.38%$, $32.2{\pm}3.22%$, $38.7{\pm}4.09%$ and $47.8{\pm}5.58%$ and there were significantly differences among groups (P<0.05). SF in Ad-SIRT6 group was lower than Ad-null and control groups after 4~10Gy X-ray irradiation (P<0.05) and the sensitization enhancement ratio (SER) was 1.35 when compared with control group. Moreover, after 48 h of 4Gy X-ray irradiation, there appeared a significant increase in G1-phase cell proportion, upregulated expression of the level of apoptosis-promoting genes (Bax and Cleaved caspase-3), but a obvious decline in S-phase and G2-phase cell proportion and a significant decrease of the level of apoptosis-inhibiting gene (Bal-2) in the Ad-SIRT6 group (P<0.05). Conclusion: The over-expression of adenovirus-mediated SIRT6, which has radiosensitization effect on A549 cells of NSCLC, can inhibit the proliferation of A549 cells and cause G0/G1 phase retardation as well as induce apoptosis of cells.