• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6349-6355
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• 2012

Previous studies showed that Math1 homologous to human Hath1 can cause mouse goblet cells to differentiate. In this context it is important that the majority of colon cancers have few goblet cells. In the present study, the potential role of Hath1 in colon carcinogenesis was investigated. Sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa, mucosa adjacent colon cancer and colon cancer samples from 48 patients. Hath1 and Muc2 expression in these samples were tested by immunohistochemistry, quantitative real-time reverse transcription -PCR and Western blotting. After the recombinant plasmid, pcDNA3.1(+)-Hath1 had been transfected into HT29 colon cancer cells, three clones were selected randomly to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time reverse transcription-PCR and Western blotting. Moreover, the proliferative ability of HT29 cells introduced with Hath1 was assessed by means of colony formation assay and xenografting. Expression of Hath1, Muc2, cyclin D1 and p27 in the xenograft tumors was also detected by Western blotting. No goblet cells were to be found in colon cancer and levels of Hath1 mRNA and Hath1, Muc2 mRNA and Muc2 were significantly down-regulated. Hath1 could decrease cyclin D1, increase p27 and Muc2 in HT29 cells and inhibit their proliferation. Hath1 may be an anti-oncogene in colon carcinogenesis.

• BMB Reports
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• 제41권7호
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• pp.529-536
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• 2008

Cinnamate-4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway, which is responsible for synthesizing a variety of secondary metabolites that participate in development and adaptation. In this study, we isolated a full-length cDNA of the C4H gene from the Korean black raspberry (Rubus sp.) and found that this gene existed as a single gene. By comparing the deduced amino acid sequence of Rubus sp. C4H with other sequences reported previously we determined that this sequence was highly conserved among widely divergent plant species. In addition, quantitative real time PCR studies indicated that the C4H gene had a differential expression pattern during fruit development, where gene expression was first detected in green fruit and was then remarkably reduced in yellow fruit, followed by an increase in red and black fruit. To investigate the two peaks in expression observed during fruit development and ripening, we measured the flavonoid content. The content of the major flavanol of Korean black raspberry fruits was determined to be highest at the beginning of fruit development, followed by a gradually decrease according to the developmental stages. In contrast, the content of anthocyanins during the progress of ripening was dramatically increased. Our results suggest that the C4H gene in Korean black raspberry plays a role during color development at the late stages of fruit ripening, whereas the expression of C4H gene during the early stages may be related to the accumulation of flavanols.

• 대한화장품학회지
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• 제39권1호
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• pp.75-81
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• 2013

Chrysin (5,7-dihydroxyflavone)은 프로폴리스, 꿀 같은 음식과 다양한 식물에 존재하는 천연 플라보노이드이다. Chrysin은 항산화, 항노화, 항염, 항암 효과 등 다양한 생물학적 효과를 가진다고 알려져 있다. 이 연구에서, 우리는 사람의 각질형성세포에서 chrysin이 VDR을 통한 transcriptional activity에 미치는 영향을 dual-luciferase assay을 통하여 살펴보았다. Chrysin은 농도 의존적으로 VDR을 통한 transcriptional activity를 증가시켰다. Quantitative real time PCR을 통해 chrysin이 사람의 각질형성세포에서 VDR mRNA의 발현을 증가시킴을 확인하였다. 또한, chrysin이 각질형성세포의 분화 마커인 keratin 10, involucrin 그리고 filaggrin의 mRNA 발현을 증가시킴을 확인하였다. 이러한 연구 결과는 chrysin이 VDR을 통한 transcriptional activity를 조절하여 각질형성세포의 분화를 촉진시킬 수 있다는 것을 시사한다.

• 한국발생생물학회지:발생과생식
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• 제21권4호
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• pp.379-389
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• 2017

Connexin (Cx) involves in the regulation of various physiological functions of tissue by forming a channel, a gap junction which allows direct cell-cell communication, between adjacent cells. The effect of a single subcutaneous treatment of estradiol benzoate (EB) or flutamide (Flu) at the weaning age on the expression of Cx isoforms in the adult caput epididymis was evaluated in this research. Using quantitative real-time PCR analysis, a low-dose of EB [$0.015{\mu}g/kg$ body weight (BW)] caused significant decreases of Cx30.3, Cx32, Cx40, Cx43, and Cx45 mRNA levels and no change of Cx26, Cx31, Cx31.1, Cx37 transcript levels. The treatment of a high-dose EB ($1.5{\mu}g/kg\;BW$) resulted in reduced expression of Cx30.3, Cx31, Cx43, and Cx45 but increased expression of Cx37 and Cx40. Expression of all Cx isoforms examined, except Cx31, was significantly increased by the treatment of a low-dose Flu ($500{\mu}g/kg\;BW$). However, the treatment of a high-dose Flu (5 mg/kg BW) led significant expressional suppression of Cx30.3, Cx31, Cx31.1, Cx32, Cx40, Cx43, and Cx45 but an increase of Cx37 transcript level. With the comparison of previous findings, the expression of Cx isoforms in the adult epididymis after the exposure to EB or Flu is likely differentially regulated in regional-specific and/or exposed postnatal age-specific manner.

• 한국발생생물학회지:발생과생식
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• 제22권1호
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• pp.29-38
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• 2018

In the multicellular tissue, cell-cell interaction is important for a precise control of its function. The exchange of signaling molecules between adjacent cells via connexon allows the functional harmony of cells in the tissue. The present research was to determine the presence and expressional patterns of connexin (Cx) isoforms in the rat epididymal fat during postnatal development using quantitative real-time polymerase chain reaction (PCR) analysis. Of 13 Cx isoforms examined, expression of 11 Cx isoforms in the epididymal fat during postnatal development was detected. These Cx isoforms include Cx26, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Expressional levels of all Cx isoforms at 1 and 2 years of age were significantly higher than those at the early postnatal ages, such as 7 days, 14 days, and 24 days of ages. Except Cx33 and Cx43, the transcript levels of rest Cx isoforms at 1 year of age were significantly lower than that at 2 years of age. In addition, expressional patterns of Cx isoforms between 7 days and 5 months of ages generally varied according to the isoform. The existence of various Cx isoforms in the rat epididymal fat has been identified and expression of each Cx isoform in the epididymal fat during postnatal development has shown a particular pattern, distinguishable from the others. To our knowledges, this is the first report showing expressional patterns of Cx isoforms at transcript level in the epididymal fat at various postnatal ages.

• 대한의생명과학회지
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• 제28권1호
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• pp.42-49
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• 2022

Tetrodotoxin (TTX) is a natural neurotoxin found in several species of puffer fish belonging to Tetraodon fugu genus and has been reported to affect processes such as proliferation, metastasis and invasion of various cancer cells. However, it was not revealed which genes were influenced by these reactions. In this experiment, it was examined in human SW620 colorectal cancer cells. The proliferation of SW620 cells was significantly reduced when treated with 0, 1, 10 and 100 μM TTX for 48 h. It was confirmed using Annexin V-propidium iodide staining that some apoptosis was induced. Differentially expressed genes (DEGs) affecting cell proliferation through RNA sequencing (RNA-seq) were selected. The expression change of DEGs was confirmed by conducting quantitative real-time polymerase chain reaction (qRT-PCR). As a result, the mRNA expression of FOS and WDR48 genes was found to be increased in the 100 μM TTX treatment group compared to the control group. On the other hand, the mRNA expression of ALKBH7, NDUFA13, RIPPLY3 and SELENOM genes was found to be reduced, and in the case of the ALKBH7 gene was identified to show significant differences. This experiment suggests that TTX can be used as an important fundamental data to elucidate the mechanism that inhibits the proliferation of SW620 cells.

• 한국어병학회지
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• 제36권2호
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• pp.229-238
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• 2023

In this study, the correlation between clinical changes and RSIV genome copy number was investigated to determine the quantitative criteria for the characteristics of RSIV infection. The rock bream (Oplegnathus fasciatus) was intraperitoneally injected with three different doses (1.0×10¹, 1.0×10³ and 1.0×10⁵ viral genome copies/fish) as low, medium, and high doses, respectively. The clinical signs (spleen enlargement, death) observation and real-time PCR were conducted at 5, 10 and 14 days post-injection. During the experiment, spleen index as a quantitative indicator for spleen enlargement was continuously increased in the medium- (up to 2.26) and high-dose (up to 4.99) challenge groups, respectively. Notably, when the spleen index was over 1.5, 2.0, 2.5 and 3.0, a positive correlation was revealed with average viral genome copy numbers of 2.51, 3.37, 4.97 and 5.43×10⁷ viral genome copies/mg, respectively. Moreover, the threshold of spleen index over 1.5 was 1.0×10⁶ viral genome copies/mg, while the thresholds of spleen index over 2.0 and dead was 2.51×10⁷ viral genome copies/mg and the thresholds of spleen index over 2.5 and 3 was 3.98×10⁷ viral genome copies/mg. These findings suggest the possibility of quantitatively analyzing the characteristics and development process of RSIV infection.

• Pediatric Infection and Vaccine
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• 제14권2호
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• pp.171-178
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• 2007

목 적 : EBV 감염은 발생 연령에 따라 혈청학적으로 진단되지 않을 수 있으며, 이 때 실시간 중합효소연쇄반응이 대신할 수 있는 진단 방법이다. 본 연구는 임상적으로 EBV 감염이 의심되는 환자에서 각각 혈청학적 검사 양성인 경우와 실시간 중합효소연쇄반응 양성인 그룹간의 임상양상을 비교하고자 하였다. 방 법 : 2004년 1월부터 2006년 12월까지 EBV 감염이 의심되는 45명의 환자를 대상으로 EBV 실시간 중합효소연쇄반응 검사를 시행하여 EBV 바이러스혈증의 존재 여부를 확인하였다. $102.5copies/{\mu}g$ DNA를 EBV 연관질병의 결정 수준으로 삼았다. 결 과 : EBV 감염이 혈청학적으로 진단된 환자는 4명이었으며 실시간 중합효소연쇄반응으로 EBV 바이러스혈증이 확인된 환자는 15명이었다. EBV 바이러스혈증이 있는 환자군의 나이는 4세 미만이 73%(11/15)였고 남녀비는 1:1.3 이었다. 입원 시의 임상증상은 발열이 73%(11/15), 인두 발적이 67%(10/15)에서 보였으며 동반된 질환으로는 급성 위장관염과 폐렴이 각각 20%(3/15)였다. 비정형 림프구가 15% 이상인 경우는 27%(4/15)에서 보였고 AST, ALT의 증가가 각각 87%(13/15), 80%(12/15)였다. CRP가 2 mg/dL 이하인 경우는 87%(13/15)였다. EBV 바이러스혈증 환자중 1세 미만은 5명으로 이들 중 80%(4/5)에서 발열이 동반되었고 비정형 림프구는 보이지 않았으며 모두 AST, ALT가 증가하였다. 혈청학적으로 진단된 환자군의 나이는 1-6세였고 75%(3/4)에서 간비대와 비장비대가 보였으며 50%(2/4)에서 발열, 인두발적, 경부 림프절 종대가 보였다. 동반된 질환은 없었으며 백혈구가 모두 $20,000/mm^3$ 이상이었고 50%(2/4)에서 비정형 림프구가 증가했으며 AST, ALT가 50 IU/L 이상으로 상승하였다. CRP는 모두 1 mg/dL 이하였다. 결 론 : 혈청학적 검사에서 진단되지 않았더라고 실시간 중합효소연쇄반응에서 EBV 바이러스혈증을 보인 환자는 EBV 감염의 일반적인 임상증상을 보였다. 따라서 EBV 감염이 의심되는 환자, 특히 1세 미만의 환자는 혈청학적 검사가 음성이어도 실시간 중합효소연쇄반응을 통해 바이러스 혈증을 확인하는 것이 EBV 감염의 진단률을 높이는데 의미가 있을 것으로 사료된다.

• 생명과학회지
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• 제24권8호
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• pp.843-850
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• 2014

본 연구에서는 전통주 제조시 부산물로 생산되는 주박과 누룩으로부터 추출물과 유기용매 분획물을 총 85종을 제조하고, 이들에 의한 항염증 활성을 연구하였다. 85종의 분획물 중 선별한 세 가지의 분획물(KSD-E1-3, KSD-E2-3, KSD-E4-3)에 의해서 LPS에 의해 염증이 유도된 RAW 264.7 세포주에서 nitric oxide 생산이 현저히 감소됨을 확인하였다. 또한, 세가지 분획물에 의해 염증유발 유전자인 COX-2, TNF-alpha, 그리고 iNOS 유전자의 발현이 감소되었다. 세 가지 분획물 중 KSD-E4-3에 의한 항염증 활성의 작용기전을 이해하기 위하여 oligo DNA microarray를 수행하였다. 마이크로어레이 결과 발현이 감소된 유전자 중 염증과 관련된 유전자 6개(IL-1F6, iNOS, IL-10, Fabp4, IL-1RN, CSF2)를 선택하여, RT-PCR과 정량적 real-time PCR을 수행하였다. 그 결과, 모든 유전자의 발현이 감소됨을 확인하였다. 결론적으로, 이러한 연구결과는 전통주 주박이 항염증 활성을 가지고 있는 식품이나 약품을 개발하는데 필요한 새로운 자원으로서 활용 가능함을 시사하는 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9853-9857
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• 2014

Background: The morbidity and mortality rate of liver cancer continues to rise in China and advanced cases respond poorly to chemotherapy. Ribosomal protein L24 has been reported to be a potential therapeutic target whose depletion or acetylation inhibits polysome assembly and cell growth of cancer. Materials and Methods: Total RNA of cultured amycin-resistant and susceptible HepG2 cells was isolated, and real time quantitative RT-PCR were used to indicate differences between amycin-resistant and susceptible strains of HepG2 cells. Viability assays were used to determine amycin resistance in RPL24 transfected and control vector and null-transfected HepG2 cell lines. Results: The ribosomal protein L24 transcription level was 7.7 times higher in the drug-resistant HepG2 cells as compared to susceptible cells on quantitative RT-PCR analysis. This was associated with enhanced drug resistance as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions: The ribosomal protein L24 gene may have effects on drug resistance mechanisms in hepatocellular carcinoma HepG2 cells.