• Molecules and Cells
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• 제38권12호
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• pp.1111-1117
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• 2015

Toll-like receptor 5 (TLR5) is a specific receptor for microbial flagellin and is one of the most well-known receptors in the TLR family. We reported previously that TLR5 signaling is well maintained during aging and that caveolin-1 may be involved in TLR5 signaling in aged macrophages through direct interactions. Therefore, it is important to clarify whether caveolin-1/TLR5 interactions affect TLR5 expression during aging. To assess the effect of caveolin-1 on TLR5, we analyzed TLR5 expression in senescent fibroblasts and aged tissues expressing high levels of caveolin-1. As expected, TLR5 mRNA and protein expression was well maintained in senescent fibroblasts and aged tissues, whereas TLR4 mRNA and protein were diminished in those cells and tissues. To determine the mechanism of caveolin-1-dependent TLR5 expression, we examined TLR5 expression in caveolin-1 deficient mice. Interestingly, TLR5 mRNA and protein levels were decreased dramatically in tissues from caveolin-1 knockout mice. Moreover, overexpressed caveolin-1 in vitro enhanced TLR5 mRNA through the MAPK pathway and prolonged TLR5 protein half-life through direct interaction. These results suggest that caveolin-1 may play a crucial role in maintaining of TLR5 by regulating transcription systems and increasing protein half-life.

• 한국콘텐츠학회논문지
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• 제20권6호
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• pp.124-130
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• 2020

본 연구에서는 시스플라틴과 백작약 에틸아세테이트 분획물의 병용 처리에 의한 암세포 성장억제 및 PMA에 의해 유도된 MMP-2 및 MMP-9 암전이 억제 효과를 조사하였다. 세포생존율 측정은 MTS법에 의해 조사하였고, MMP-2/-9의 유전자발현과 활성은 RT-PCR과 Zymography법을 통하여 확인하였다. 결과에 의하면, 백작약, 시스플라틴의 농도가 증가함에 따라 세포 성장억제 효과가 증가함을 보였다. 또한, 단독 처리에 비해 200 μM의 시스플라틴과 50 ㎍/ml의 백작약 병용 처리에 의해서는 YD-10B 세포의 성장이 50% 감소하였다. PMA 처리된 YD-10B 세포에서 50 ㎍/ml의 백작약과 200 μM의 시스플라틴을 병용 처리하였을 때, MMP-2 및 MMP-9의 mRNA 발현과 단백질 활성들이 모두 유의하게 억제하였다. 그러므로 본 연구에서는 시스플라틴과 백작약의 병용 처리는 시스플라틴 단독 처리보다 구강암의 암 침윤을 억제할 수 있는 효과적인 항암제로서의 가능성을 기대할 수 있다.

• The Journal of Korean Physical Therapy
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• 제22권3호
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• pp.71-77
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• 2010

Purpose: To determine whether upregulation of inducible nitric oxide synthase (iNOS) transcription and translation is related to radicular pain in a model of lumbar disc herniation. Also, to investigate the temporal changes of mRNA expression of iNOS and the identity of iNOS and transient receptor potential vanilloid (TRPV) 1 channel expression cells in dorsal root ganglion (DRG) of a model of lumbar disc herniation. Methods: A lumbar disc herniated rat model was developed by implantation of the autologous nucleus pulposus, harvested from the coccygeal vertebra of each tail, on the left L5 nerve root just proximal to the DRG. Rats were tested for mechanical allodynia of the plantar surface of both hind paws 2 days before surgery and 1, 5, 10, 20 and 30 days postoperatively. Reverse transcription polymerase chain reaction (RT-PCR) was used to follow iNOS mRNA expression. To stain iNOS and TRPV1 in DRG, an immunohistochemical study was done 10 days after surgery. Results: A significant drop in mechanical withdrawal threshold on the ipsilateral and contralateral hind paws was observed 1 day after surgery and was prolonged to 30 days in rats with lumbar disc herniation. The expression of mRNA for iNOS peaked at postoperative day 10 on both sides of the DRG. iNOS-positive sensory neurons in the DRG varied in size from large to small diameter cells. A majority of small and intermediate sensory neurons were TRPV1-positive cells. Double immunofluorescence staining for TRPV1 and iNOS revealed that most intermediate TRPV1-positive sensory neurons co-localized with iNOS-positive neurons. Conclusion: Nucleus pulposus-induced mechanical allodynia can be generated without mechanical compression. This pain is related to temporal changes in expression of iNOS mRNA in the DRG. Co-localization of TRPV1 and iNOS in intermediate neurons of the DRG is correlated with pain modality and intensity.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1639-1642
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• 2014

The multidrug resistance phenotype is one of the major problems in development of cancer cell resistance to chemotherapy. Some natural compounds from medicinal plants have demonstrated promising capacity in enhancing anticancer effects in drug resistant cancer cells. We aimed to investigate whether mangiferin might have an ability to re-sensitize MCF-7 breast cancer cells previously treated with short-term doxorubicin in vitro, through the modulation of efflux transporters, P-glycoprotein (P-gp), MRP1 and BCRP. We exposed MCF-7 breast cancer cells pretreated with doxorubicin for 10 days to mangiferin (10, 25 or 50 ${\mu}M$) for 96 hours. Afterwards, we evaluated influence on cell viability and level of mRNA expression of P-gp, MRP1 and BCRP. Doxorubicin given in combination with mangiferin at low concentrations (10 and 25 ${\mu}M$) failed to give significant reduction in cell viability, while at the highest concentrations, the combination significantly reduced cell viability. The mRNA expression analysis of P-gp, MRP1 and BCRP showed that mangiferin had inhibitory effects on P-gp but no effects on MRP1 and BCRP. In conclusion, we suggest that mangiferin at high concentrations can be used as chemosensitizer for doxorubicin therapy. This effect might be attributed by inhibitory effects of mangiferin on P-glycoprotein expression.

• Asian Pacific Journal of Cancer Prevention
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• 제14권6호
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• pp.3503-3507
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• 2013

Background: Although the majority of investigations concerned with TP53 and its protein have focused on coding regions, recently a set of studies highlighted significant roles of regulatory elements located in p53 mRNA, especially 5'UTR. The wrap53${\alpha}$ transcript is one of those that acts as a natural antisense agent, forming RNA-RNA hybrids with p53 mRNA and protecting it from degradation. Materials and Methods: In this study, we focused on the mutation status of exon $1{\alpha}$ of the WRAP53 gene (according to exon 1 of p53) in 160 breast tumor tissue samples and conducted a bioinformatics search for probable miRNA binding site in the p53/wrap53 overlapping region. Mutations were detected, using single stranded conformation polymorphism (SSCP) and sequencing. We applied the miRBase database for prediction of miRNAs which target overlapping region of p53/wrap53 transcripts. Results: Our results showed all samples to have wild type alleles in exon 1 of TP53 gene. We could detect a novel and unreported intronic mutation (IVS1+56, G>C) outside overlapping regions of p53/wrap53 genes in breast cancer tissues and also predict the presence of a binding site for miR-4732-5p in the 5'UTR of Wrap53 mRNA. Conclusions: From our findings we propose designing further studies focused on overexpression of miRNA-4732-5p and introducing different mutations in the overlapping region of wrap53 and p53 genes in order to study their effects on p53 and its ${\Delta}N$ isoform (${\Delta}$40p53) expression. The results may provide new pieces in the p53 targeting puzzle for cancer therapy.

• The Journal of Korean Physical Therapy
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• 제15권4호
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• pp.190-198
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• 2003

Expression of c-fos, an immediate early gene, has accepted to be a marker of functional activity in neurons. This study was aimed to investigate the effects of cold therapy on the expression of spinal cord c-fos in rats of carrageenan-induced muscle pain. Muscle pain was induced in male Sprague-Dawley rats by intra-muscular injection of gastrocnemius with $2\%$ carrageenan. The paw withdrawal latency (PWL) and tail flick test (TFT) responses to heat stimuli were used to detect secondary hyperalgesia produced by the muscle pain and measured to assess the effects of cold. The expression of c-fos was determined in the lumbar regions of the spinal cord by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry assays. The secondary hyperalgesia to heat simuli (PWL and TFT) were significantly reduced in cold therapy compared with that in the controls. In RT-PCR assays the expression of c-fos mRNA was down-regulated in the lumbar spinal cord in cold group. In addition, Fos immunoreactivity in the dorsal horn of the lumbar spinal cord was decreased in cold group. These results suggested that application of cold attributed to increase PWL and TFT responses and to decrease expression of the c-fos produced by muscle pain.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5391-5396
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• 2013

Aim: To investigate the role of Golgi phosphoprotein 3 (GOLPH3) in tumour growth and metastasis of esophageal squamous cancer. Methods: A lentiviral shRNA-vector was utilized to stably knockdown GOLPH3 in Eca-109 esophageal squamous cancer cells. mRNA transcription and protein expression of GOLPH3 were examined by real-time quantitative PCR and Western blotting, respectively. Cell proliferation activity was assessed by MTT assay and invasion and migration potentials by matrigel invasion and transwell motility assays. Results: Stable knockdown in the GOLPH3 cell line was established. PD-A gene expression was significantly suppressed by lentivirus-mediated RNAi, which resulted in reducing the capacity for cell proliferation, migration, invasion and adhesion in vitro. In vivo, GOLPH3 depletion resulted in inhibition of tumour growth, with stable decrease in the expression of GOLPH3 in tumor xenografts. Conclusions: Our findings suggest that lentivirus mediated silencing of the GOLPH3 gene has a significant anti-tumour effect on esophageal squamous cancer in vitro and in vivo. In addition, the results indicate that GOLPH3 might be an effective molecular target for gene therapy in esophageal squamous cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제13권2호
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• pp.473-477
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• 2012

Introduction: The esophagus squamous cell carcinoma (ESCC) is one of the most deadly malignances, and a current challenge is the development of effective therapeutic agents. Our present work addressed the effect of HIF-$1{\alpha}$ siRNA alone or in combination with cisplatin on the growth of ESCC in nude mice. Materials and Methods: Xenografts were established by inoculating ESCC TE-1 cells in nude mice, and transplanted tumors were treated with HIF-$1{\alpha}$ siRNA, cisplatin alone or together. Growth was assessed by measuring tumor volume. HIF-$1{\alpha}$ mRNA and protein expression were detected using RT-PCR and immunohistochemistry, respectively. Apoptosis of ESCC TE-1 cells was analyzed by flow cytometry. Results: In our nude mice model, HIF-$1{\alpha}$ siRNA effectively inhibited the growth of transplanted ESCC, downregulating HIF-$1{\alpha}$ mRNA and protein expression, and inducing ESCC TE-1 cell apoptosis. Notably when combinated with cisplatin, HIF-$1{\alpha}$ siRNA showed synergistic interaction in suppressing tumor growth. Furthermore, the proportion of apoptotic cells in HIF-$1{\alpha}$ siRNA plus cisplatin group was significantly higher than that in cisplatin or HIF-$1{\alpha}$ siRNA-treated groups (P<0.05). Conclusions: Down-regulated HIF-$1{\alpha}$ expression induced by siRNA could effectively suppress the growth of transplanted ESCC $in$ $vivo$. HIF-$1{\alpha}$ siRNA could enhance the cytotoxicity of cisplatin, which suggests that a combination of these two agents may have potential for therapy of advanced ESCC.

• Nuclear Medicine and Molecular Imaging
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• 제41권3호
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• pp.209-217
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• 2007

목적: mdr1유전자를 표적으로 한 short hairpin RNA (shMDR)는 다약재내성을 나타내는 암세포에서 효과적으로 mdr1 유전자의 발현을 억제 할 수 있고 sodium iodide symporter (NIS)는 유전자 치료와 리포터로의 기능을 동시에 나타낼 수 있다. 이 연구에서는 사람 대장암세포(HCT15)에 shMDR과 NIS를 동시에 이입하고 Tc-99m sestamibi와 I-125 섭취를 측정하였고 doxorubicin과 I-131 치료효과도 관찰하였다. 대상 및 방법: 사람 태아 신장 세포주(Human Embryonic Kidney cells; HEK293)에 liposome 시약으로 shMDR을 이입하고 RT-PCR과 western blot으로 분석하였다. shMDR와 NIS 유전자가 발현하는 adenovirus를 만들고 HCT15 세포에 이입 후 48시간에 shMDR에 의한 Pgp의 기능 억제를 확인하기위해 Tc-99m sestamibi 섭취와 doxorubicin 세포독성을 측정하였다. 또한 NIS유전자의 기능을 확인 하기위해 I-125 섭취와 I-131 세포독성도 확인하였다. 결과: shMDR이 이입 된 HEK293 세포에서 mdr1의 mRNA와 Pgp의 발현이 각각 75%, 80% 감소하였다. NIS 유전자가 발현하는 adenovirus를 HCT15 세포에 이입하고 NIS 유전자 발현을 확인 한 결과 대조군에 비해 월등히 높게 발현하였다. Ad-shMDR 300 MOI, Ad-shMDR 300 MOI 와 Ad-NIS 10 MOI를 처리한 경우 Tc-99m sestamibi의 섭취가 대조군보다 1.5배 정도 증가하였다. HCT15 세포에 Ad-NIS 10 MOI를 감염시킨 경우 I-125 섭취가 대조군에 비해 25배 이상 증가였다. 또한 Ad-shMDR와 Ad-NIS를 동시 감염 시켰을 경우 doxorubicin의 세포 독성이 증가하여 나타났고 Ad-NIS 20 MOI를 감염시켰을 때 I-131에 의한 세포독성이 대조군보다 증가하였다. 결론: 세포에 shMDR의 이입으로 mdr1 유전자의 발현이 억제되고 Tc-99m sestamibi의 섭취와 doxorubicin의 세포독성이 증가하였으며 NIS 유전자의 이입으로 I-125의 섭취와 I-131의 세포독성이 증가하였다. 다약제내성세포에 shMDR와 NIS 유전자의 동시 이입은 doxorubicin과 방사성 옥소의 이중치료 효과를 높일 수 있을 것으로 본다.

• Clinical and Experimental Pediatrics
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• 제57권3호
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• pp.117-124
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• 2014

Purpose: Administration of antiretroviral drugs to mothers and infants significantly decreases mother-to-child human immunodeficiency virus (HIV) transmission; cesarean sections and discouraging breastfeeding further decreases this risk. The present study confirmed the HIV status of babies born to mothers infected with HIV and describes the characteristics of babies and mothers who received preventive treatment. Methods: This study retrospectively analyzed medical records of nine infants and their mothers positive for HIV who gave birth at Korea University Ansan Hospital, between June 1, 2003, and May 31, 2013. Maternal parameters, including HIV diagnosis date, CD4+ count, and HIV ribonucleic acid (RNA) copy number, were analyzed. Infant growth and development, HIV RNA copy number, and HIV antigen/antibody test results were analyzed. Results: Eight HIV-positive mothers delivered nine babies; all the infants received antiretroviral therapy. Three (37.5%) and five mothers (62.5%) were administered single- and multidrug therapy, respectively. Intravenous zidovudine was administered to four infants (50%) at birth. Breastfeeding was discouraged for all the infants. All the infants were negative for HIV, although two were lost to follow-up. Third trimester maternal viral copy numbers were less than 1,000 copies/mL with a median CD4+ count of $325{\mu}L$ ($92-729{\mu}L$). Among the nine infants, two were preterm (22.2%) and three had low birth weights (33.3%). Conclusion: This study concludes that prophylactic antiretroviral therapy, scheduled cesarean section, and prohibition of breastfeeding considerably decrease mother-to-child HIV transmission. Because the number of infants infected via mother-to-child transmission may be increasing, studies in additional regions using more variables are necessary.