• 한국동물생명공학회지
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• 제37권2호
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• pp.121-129
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• 2022

Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNPA2/B1) is an N6-methyladenosine (m⁶A) RNA modification regulator and a key determinant of prem-RNA processing, mRNA metabolism and transportation in cells. Currently, m⁶A reader proteins such as hnRNPA2/B1 and YTHDF2 has functional roles in mice embryo. However, the role of hnRNPA2/B1 in porcine embryogenic development are unclear. Here, we investigated the developmental competence and mRNA expression levels in porcine parthenogenetic embryos after hnRNPA2/B1 knock-down. HhnRNPA2/B1 was localized in the nucleus during subsequent embryonic development since zygote stage. After hnRNPA2/B1 knock-down using double stranded RNA injection, blastocyst formation rate decreased than that in the control group. Moreover, hnRNPA2/B1 knock-down embryos show developmental delay after compaction. In blastocyste stage, total cell number was decreased. Interestingly, gene expression patterns revealed that transcription of Pou5f1, Sox2, TRFP2C, Cdx2 and PARD6B decreased without changing the junction protein, ZO1, OCLN, and CDH1. Thus, hnRNPA2/B1 is necessary for porcine early embryo development by regulating gene expression through epigenetic RNA modification.

• Journal of Nutrition and Health
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• 제42권6호
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• pp.505-515
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• 2009

본 연구에서는 비타민 C, silicon, 철분을 농도별로 피부섬유아 세포에 처리 후 콜라겐 합성 효소인 PH, LH의 mRNA와 protein 발현 및 분해 효소인 MMP-1와 저해제인 TIMP-1의 mRNA, protein 발현의 변화를 대조군과 비교 분석하였으며 그 결과를 요약하면 다음과 같다. 1) 피부 섬유아세포에서 비타민 C의 처리는 콜라겐 합성 효소인 PH의 mRNA 발현을 대조군에 비해 현저하게 증가시켰고 이와 병행하여, PH의 protein 발현도 대조군에 비해 유의적으로 증가하였다. 그러나 다른 콜라겐 합성 효소인 LH의 mRNA 발현에는 영향을 미치지 않았으나 protein의 발현을 증가시켰다. 또한, 콜라겐 분해 효소인 MMP-1의 mRNA 발현은 대조군에 비해 유의적으로 증가시켰으나 protein 발현에서는 대조군과 차이가 없었다. 2) 피부 섬유아세포에서 silicon의 혈중 내 농도 처리는 LH의 mRNA 발현의 현저한 증가와 더불어 protein 발현에도 긍정적인 영향을 미치는 것으로 나타났다. 콜라겐 분해 효소인 MMP-1과 저해제인 TIMP-1의 단백질 발현을 대조군에 비해 증가시켰다. 3) 피부 섬유아세포에서 철분의 혈중 내 농도 처리는 콜라겐 합성 및 분해 관련 효소의 mRNA 및 protein 발현에 영향을 미치지 않았다. 결론적으로, 피부섬유아세포에서 비타민 C 및 silicon의 처리는 콜라겐의 posttranslational modification 관련 효소의 mRNA의 발현 및 단백질의 발현을 증가시켜 궁극적으로 콜라겐 합성에 긍정적인 영향을 나타내었다.

• Toxicological Research
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• 제20권1호
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• pp.31-36
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• 2004

Against environmental stress, ceruloplasmin which is a plasma protein, are believed to play central roles in antioxidant- or peroxidase-activity in blood stream to remove free radicals, which may be caused by exposing of $\gamma$-irradiation. In human U-937 cells exposed to $\gamma$-irradiation, the levels of mRNA in ceruloplasmin gene were measured on 0, 4, 12, 24 hr after exposing by using comparative RT-PCR (Reverse transcriptase-polymerase chain reaction) which was achieved to compare with house keeping genes such as $\beta$-actin and hprt. After $\gamma$-irradiation of 100 rads or 200 rads, the total quantities of RNA were increased as dose and time dependent manner. On the contrary, the variation of mRNA expression in ceruloplasmin was not found until 4 hr after irradiation. After 12 hr and 24 hr of irradiation, the levels of mRNA in ceruloplasmin were significantly increased as dose and time dependent manner than un-exposed cells.

• Imaging Science in Dentistry
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• 제34권2호
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• pp.99-106
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• 2004

Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, especially on the osteonectin and bone sialoprotein. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1, 4 and 8 Gy at a dose rate of 5.38 Gy/min using Cs-I37 irradiator. After specimens were harvested, total RNA was extracted on the 3rd, 7th, 14th, 21st day after irradiation. The total RNA was reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR with a pair of primers. Results: The irradiated cells showed a dose-dependent increase in osteonectin mRNA expression when compared with the unirradiated control group. The irradiated cells showed no difference in bone sialoprotein mRNA expression when compared with the unirradiated control group. In accordance with the duration of culture period after irradiation, the level of osteonectin mRNA expression showed no difference, but it increased a little at the 21st day in the 4 and 8 Gy exposure groups. In the case of bone sialoprotein, however, the level of mRNA expression increased significantly at the 3rd and 7th day after irradiation, but it showed no difference at the 14th and 21st day when compared with the control group. Conclusion: These results showed that each single dose of 0.5, 1, 4 and 8 Gy influenced the mRNA expression of osteonectin and bone sialoprotein at the calcification stage of osteoblastic cells, suggesting that single dose of irradiation affected the osteoblastic bone formation at the cell level.

• 대한수의학회지
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• 제40권2호
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• pp.229-236
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• 2000

The synthesis of steroid hormone starts from cholesterol. Steroidogenic acute regulatory protein (StAR) acutely transfers cholesterol from the outer mitochondrial membrane to the inner in the early step of steroidogenesis. Many kinds of steroid hormone are mainly synthesized in adrenal grand, ovary, and testis. Among the steroid hormone, testosterone is synthesized in Leydig cells of the testis, the production of testosterone significantly increases in adult testis after puberty onset. Therefore, we think that the expression of StAR mRNA in testis will change according to the testicular development. The aim of this study is to determine the distribution of StAR mRNA in immature and adult rat testes and to confirm the functions of StAR in these testes. Thus, in situ hybridization was used in rat testes of the 2, 4, and 10 weeks of age. StAR mRNA was expressed in Leydig cells. Positive signals of StAR mRNA were weakly detected in Leydig cells of the 2 weeks of age. But, StAR mRNA was strongly expressed in Leydig cells of the 4 and 10 weeks of age, where steroidogenesis actively occur. In our results, the pattern of StAR mRNA expression was similar to the pattern of testosterone production in immature and adult rat testes. In conclusion, we can suggest that StAR acts as an important factor to regulate the synthesis of testosterone in Leydig cells of the rat testis.

• 한국축산식품학회지
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• 제43권4호
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• pp.703-711
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• 2023

As an initial study to elucidate the molecular mechanism of how probiotics modulate macrophage activity, we monitored mRNA expression patterns in peritoneal macrophages (PMs) treated with two different strains of probiotics. After treatment with either Weissella cibaria WIKIM28 or Latilactobacillus sakei WIKIM50, total RNAs from PMs were isolated and subjected into gene chip analyses. As controls, mRNAs from vehicle (phosphate-buffered saline, PBS)-treated PMs were also subjected to gene chip analysis. Compared to vehicle (PBS)-treated PMs, WIKIM28-treated and WIKIM50-treated PMs exhibited a total of 889 and 432 differentially expressed genes with expression differences of at least 4 folds, respectively. Compared to WIKIM28-treated PMs, WIKIM50-treated PMs showed 25 up-regulated genes and 21 down-regulated genes with expression differences of more than 2 folds. Interestingly, mRNA transcripts of M2 macrophage polarization marker such as anxa1, mafb, and sepp1 were increased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. Reversely, mRNA transcripts of M1 macrophage polarization marker such as hdac9, ptgs2, and socs3 were decreased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. In agreement with these observations, mRNA expression levels of tumor necrosis factor-α and interleukin-1α were significantly reduced in WIKIM50-treated macrophages compared to those in WIKIM28-treated macrophages. These results may indicate that probiotics can be classified as two different types depending on their ability to convert macrophages into M1 or M2 polarization.

• 환경생물
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• 제22권1호
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• pp.206-212
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• 2004

In an effort to develop the tools for monitoring the contamination of xenoestrogen in the aquatic environment of Korea, reverse transcription-polymerase chain reaction (RT-PCR) analysis of vitellogenin (VTG) mRNA expression were optimized in Synechogobius hastus. Based on the partial VTG cDNA sequence VTG mRNA level in livers from male fishes was analyzed by RT-PCR. As an internal control beta actin mRNA was amplified. 3 ${\mu}g$ of total RNA was reverse transcribed in 20 $\mu$l reaction using murine leukemia virus 〔MuLV〕 reverse transcriptase. Subsequent PCR using the 1 ${\mu}g$ of cDNA resulted in linear increase in PCR product of VTG in female liver cDNA from 10 to 30 cycles of amplification. On the contrary, in male, PCR product first detected at 28 cycles of amplification and linearly increased during 38 cycles of amplification, suggesting that male S. hastus expresses minute amount of VTG mRNA which is $2^{-18}$ equivalent of female. In conclusion, the optimized protocol of VTG mRNA expression in the liver of male S. hastus will be promising the environmental monitoring the xenoestrogen contamination in the western coast and estuaries in Korea.

• Genomics & Informatics
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• 제5권1호
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• pp.32-35
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• 2007

Telomerase reverse transcriptase (TERT) is an enzymatic ribonucleoprotein that prolongs the replicative life span of cells by maintaining protective structures at the ends of eukaryotic chromosomes. Telomerase activity is highly up-regulated in 85-90% of human cancers, and is predominately regulated by hTERT expression. In contrast, most normal somatic tissues in humans express low or undetectable levels of telomerase activity. This expression profile identifies TERT as a potential anticancer target. By using an RNA mapping strategy based on a trans-splicing ribozyme library, we identified the regions of mouse TERT (mTERT) RNA that were accessible to ribozymes. We found that particularly accessible sites were present downstream of the AUG start codon. This mTERTspecific ribozyme will be useful for validation of the RNA replacement as cancer gene therapy approach in mouse model with syngeneic tumors.

• 대한약침학회지
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• 제7권2호
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• pp.43-56
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• 2004

Objectives : In order to investigate effects and immune improvement of distilled red-ginseng herbal Acupuncture, expression of Cox-1, Cox-2, and mRNA of Bcl-2 and Bax were analyzed in A549 cell in vivo. Survival time and expression of cytokine mRNA were measured for the mice with Sarcoma-180 induced abdominal cancer. Methods : Balb/c mouse was treated with distilled red-ginseng Herbal Acupuncture at Wisu($BL_{21}$) and Chung- wan($CV_{12}$) to investigate anti-cancer effects and immune response. Results : 1. For expression of mRNA of Cox-1 using RT-PCR, the control group and the experiment groups didn't show significant differences. For Cox-2, both experiment groups and the normal group showed significant differences. 2. For expression of mRNA of Bcl-2 using RT-PCR, experiment groups showed slight decrease compared to the control group. For Bax, no significant changes were shown between the control group and experiment groups. 3. For survival time, all of experiment groups showed 11.1% increase compared to the control group. 4. For IL-2 and IL-4 productivity using Flow cytometry, all of experiment groups didn't show any significance. 5. For IL-2 productivity using ELISA, all of experiment groups didn't show any significance. 6. For expression of cytokine mRNA using RT-PCR, significant increase of IL.-2 and IL-4 were witnessed in the experiment group II compared to the control group. Significant increase of IL-10 was shown in all off experiment groups compared to the control group. Conclusion : According to the results, we can expect that distilled red-ginseng Herbal Acupuncture may be further effccts in anti-cancer and immune improvement if increasing concentration.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.811-816
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• 2007

Adipocyte fatty acid-binding protein (A-FABP), which belongs to the FABP family, plays an essential role in long-chain fatty acid uptake and metabolic homeostasis, especially in adipose tissue. The pattern of A-FABP gene mRNA expression in different growth stages and its relation to intramuscular fat (IMF) accretion in pigs was studied. Fifteen female $Duroc{\times}Landrace{\times}Yorkshire$ pigs in five groups of three pigs each, weighing 1, 30, 50, 70 and 90 kg were used to study developmental gene mRNA expression of A-FABP in various adipose tissues by means of semi-quantitative RT-PCR. Results showed that A-FABP mRNA levels in subcutaneous and ventral adipose tissues first increased from 1 to 50 kg, then gradually declined from 50 to 90 kg. Moreover, the rank order of A-FABP mRNA levels determined in three adipose tissues was as follows: subcutaneous adipose>ventral adipose>mesenteric adipose. A-FABP mRNA expression in mesenteric adipose tissue was constant during development. In addition, a positive correlation from 1 to 50 kg BW pigs and a negative correlation from 50 to 90 kg BW between A-FABP mRNA levels in subcutaneous and ventral adipose and IMF content were found.