• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.4065-4069
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• 2015

Background: Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein specifically expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. Epithelial-mesenchymal transition (EMT) was found to be closely related with tumor progression. Accordingly, it is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer cells. Materials and Methods: EGFL7 expression was firstly testified in 4 pancreatic cancer cell lines by real-time polymerase chain reaction (Real-time PCR) and western blot, and the highest expression of EGFL7 was found in PANC-1 cell line. Then, PANC-1 cells transfected with small interference RNA (siRNA) of EGFL7 using plasmid vector were named si-PANC-1, while transfected with negative control plasmid vector were called NC-PANC-1. Transwell assay was used to analyze the migration of PANC-1 cells. Real-time PCR and western blotting were used to detect the expression change of EGFL7 gene, EMT markers like E-Cadherin, N-Cadherin, Vimentin, Fibronectin and transcription factors like snail, slug in PANC-1, NCPANC-1, and si-PANC-1 cells, respectively. Results: After successful plasmid transfection, EGFL7 gene were dramatically knock-down by RNA interference in si-PANC-1 group. Meanwhile, migration ability decreased significantly, compared with PANC-1 and NC-PANC-1 group. Meanwhile, the expression of epithelial phenotype marker E-Cadherin increased and that of mesenchymal phenotype markers N-Cadherin, Vimentin, Fibronectin dramatically decreased in si-PANC-1 group, indicating a reversion of EMT. Also, transcription factors snail and slug decreased significantly after RNA interference. Conclusions: Current study suggested that highly-expressed EGFL7 promotes migration of PANC-1 cells and acts through transcription factors snail and slug to induce EMT, and further study is needed to confirm this issue.

• Advances in Traditional Medicine
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• 제1권1호
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• pp.8-27
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• 2000

Components of extracellular matrix and the matrix-degrading enzymes are some of the key regulators of tumor metastasis and angiogenesis. Hyaluronic acid (HA), a matrix glycosaminoglycan, is known to promote tumor adhesion and migration, and its small fragments are angiogenic. Until now, we have compared levels of hyaluronidase, an enzyme that degrade HA, in normal adult prostate, benign prostate hyperplasia and prostate cancer tissues and in conditioned media from epithelial explant cultures, using a substrate (HA)-gel assay and ELISA-like assay (Kim et al., unpublished results). The present review described an overall characterization of hyaluronidases and its application to human diseases. The hyaluronidases are a family of enzymes that have, until recently, deed thorough explication. The substrate for these enzymes, hyaluronan, is becoming increasingly important, recognized now as a major participant in basic processes such as cell motility, wound healing, embryogenesis, and implicated in cancer progression. And in those lower life forms that torment human beings, hyaluronidase is associated with mechanisms of entry and spread, e.g. as a virulence factor for bacteria, for tissue dissection in gas gangrene, as a means of treponema spread in syphilis, and for penetration of skin and gut by nematode parasites. Hyaluronidase also comprises a component of the venom of a wide variety of organisms, including bees, wasps, hornets, spiders, scorpions, sh, snakes and lizards. Of particular interest is the homology between some of these venom hyaluronidases and the enzyme found in the plasma membrane of mammalian spermatozoa, attesting to the ancient nature of the conserved sequence, a 36% identity in a 300 amino acid stretch of the enzyme protein. Clearly, hyaluronidase is of biological interest, being involved in the pathophysiology of so many important' human disorders. Greater effort should be made in studying this family of enzymes that have, until recently, been overlooked. Also, oriental medical application of the hyaluronidase will be discussed with respect to inhibition and suppression of inflammation and malignacy.

• 대한두경부종양학회지
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• 제14권2호
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• pp.228-235
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• 1998

Objectives: Papillary microcarcinoma of the thyroid was evaluated as to the effectiveness of diagnostic modalities, lymphatic spread pattern, and therapeutic decision according to tumor size. Material and Methods: We retrospectively analyzed a clinicopathologic findings of 72 papillary microcarcinoma patients who were treated at the over 11 years between 1985 and 1995. The authors divided papillary microcarcinoma of the thyroid into two subgroups according to tumor size: $0{\leqq}5mm$ and $5<0{\leqq}10mm$. An analysis including age and gender distribution, diagnostic tools(thyroid sonogram, thyroid scan, thyroid function test, fine needle aspiration cytology, frozen section), pathological examination of lymphnode, and surgical procedures was carried out in each subgroups. Results: The carcinoma of smaller than 5mm were found in 32 patients, and of 6 -10mm were in 40 patients. The average age of patients was 45years and all of them were female. Cold nodules on thyroid scan were noticed in 53 patientss and normal findings were in 15 patients. Suspicious malignant lesions(fine calcification, solid mass, irregular margin) on thyroid sonography were detected in 23 patients and the sonography was more useful in detecting $0{\leqq}5mm$ small sized lesions than other diagnostic methods. FNAC were performed in 17 patients, and 7 patients were diagnosed as having thyroid papillary cancer. But diagnotic rate in $0{\leqq}5mm$ small sized lesions was very low(one of eights).Frozen section were performed in all patients, among these 15 patients were diagnosed as being benign diseases and false negative rates were higher in $0{\leqq}5mm$ small sized lesions than in $5<0{\leqq}10mm$ sized lesions(p-value<0.006). Only thyroidectomies were performed in 24 patients and thyroidectomy with node dissections in 48 patients. The lymphnode metastatic rates were much higher in multifocal lesions(61.5%) than in single lesion. The incidence of cervical lymphnode metastasis was 19.4% in $0{\leqq}5mm$ sized lesions and 47.9% in $5<0{\leqq}10mm$ sized lesions. Postoperative management were performed with TSH suppression therapy(T4, synthroid) in all patients and RI therapy in 29 patients. Conclusion: On the basis of our study, improved preoperative diagnostic tools for papillary microcarcinoma of the thyroid was helpful in the choice of surgical treatment. As a result of techninological progress(ultrasonography, FNAC), the pencentage of the discovery of papillary microcarcinoma has been increased. The thyroid ultrasonography was useful in detecting small sized lesions($0{\leqq}5mm$), but FNAC may not be beneficial in detecting small sized lesions($0{\leqq}5mm$). In the surgical procedure, thyroid lobectomy alone should be avoided because of the high rate of bilaterality and multifocality.

• 생명과학회지
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• 제16권6호
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• pp.964-971
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• 2006

기질의 침윤과 전이를 특징으로 하는 악성종양 세포는 세포외 기질이나 기저막에 의존적으로 작용한다. 세포외 기질을 분해하는 효소인 matrix metalloproteinase (MMP) 계들의 발현 및 활성증가는 대부분의 악성종양세포에서 전이와 침윤을 촉진시킨다. MMP family 가운데 특히 type IV collagenase 활성을 지닌 MMP-2와 MMP-9은 세포외기질의 중요한 구성분인 collagen, fibronectin을 분해하는 특성을 가지며 암 전이를 용이하게 하는 주요한 효소로 잘 알려져 있다. 본 연구에서는 항암후보물질인 disulfiram이 골 육종 (U20S), 신장암 (Caki-1) 및 자궁암 (Caski) 세포에서 MMP-2와 MMP-9의 효소활성 및 발현억제에 대해 조사하였다. MTT assay를 이용하여 disulfiram에 대한 암세포 viability 실험에서는 disulfiram이 암세포의 viability를 저해하였다. 또한 zymography, western blot 및 RT-PCR 등을 이용한 type IV collagenase의 활성 및 발현 실험에서 disulfiram은 type IV collagenase의 활성을 비롯하여 단백질 및 mRNA 발현을 억제시키는 것을 확인하였다. 따라서 disulfiram이 MMP-2와 MMP-9의 활성 및 발현 억제 기전을 통하여 골 육종, 신장암 및 자궁경부암 세포의 작용을 억제한다는 연구 결과는 disulfiram이 각종 악성종양의 침윤과 전이를 억제 또는 방지하기 위한 치료물질로서 임상에서 활용할 수 있는 가능성을 보여준다.

• 한국식품저장유통학회지
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• 제24권8호
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• pp.1122-1128
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• 2017

애호박의 부위 중 덩굴손 유래의 열수 추출물에 대한 세포 독성 및 염증제어활성 대하여 조사하였다. 애호박 덩굴손의 열수에 대한 세포의 독성에 미치는 영향을 조사한 결과 0.313-2.5 mg/mL의 농도일 때 물 추출물은 세포의 생장에 영향을 미치지 않아 세포에 대한 독성이 없다고 판단되었다. BMDMs를 배양한 세포에 호박덩굴손 열수 추출물과 LPS와 함께 처리한 결과 염증성 사이토카인인 IL-6 및 TNF-${\alpha}$의 농도 의존적으로 감소함을 확인하였다. 호박 덩굴손 열수 추출물로부터 단리 정제된 rutin에 의한 염증성 사이토카인의 분비가 억제되었다. 국내에서 재배되고 있는 호박의 부위별로 조사해본 결과 호박부위 중 유일하게 덩굴손부분에서 염증억제 활성을 보였다. 쥬키니 호박, 애호박 및 늙은 호박의 덩굴손에서 유효성분인 rutin을 함유하고 있었고 향후 염증 억제 소재로서의 활용가능성을 시사하였다.

• Journal of Korean Neurosurgical Society
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• 제65권6호
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• pp.790-800
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• 2022

Objective : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.

• Advances in nano research
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• 제12권6호
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• pp.639-652
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• 2022

The prolyl 3-hydroxylase family member 4 (P3H4), is associated with post-translational modification of fibrillar collagens and aberrantly activated in cancer leading to tumor progression. However, its role in clear cell renal cell carcinoma (ccRCC) is still unknown. Here we reported that P3H4 was highly expressed in renal cancer tissues and significantly positive correlated with poor prognosis. Knockdown of P3H4 inhibited the proliferation, migration and metastasis of renal cancer cells in vitro and in vivo, and also, overexpression of it enhanced the oncogenic process. Mechanistically, P3H4 depletion decreased the levels of GDF15-MMP9 axis and repressed its downstream signaling. Further functional studies revealed that inhibition of GDF15 suppressed renal cancer cell growth and GDF15 recombinant human protein (rhGDF15) supplementation effectively rescued the inhibitory effect induced by P3H4 knockdown. Moreover, decreased levels of MMP9 caused by inhibition of P3H4-GDF15 signaling constrained the expression of PD-L1 and suppression of P3H4 accordingly promoted anti-tumor immunity via stimulating the infiltration of CD4+ and CD8+ T cells in syngeneic mice model. Taken together, our findings firstly demonstrated that P3H4 promotes ccRCC progression by activating GDF15-MMP9-PD-L1 axis and targeting P3H4-GDF15-MMP9 signaling pathway can be a novel strategy of controlling ccRCC malignancy.

• 농업생명과학연구
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• 제50권2호
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• pp.151-160
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• 2016

염증성 사이토카인은 파골세포형성과정에서 중요한 요인이며, 뼈의 흡수는 자주 골다공증과 연결된다. 설포라판은 보로콜리의 화뢰로 부터 분리된 물질로 염증성 사이토카인을 억제한다고 알려져 있다. 본 실험에서는 Receptor activator of nuclear factor kappaB ligand(RANKL)로 자극된 세포에서 설포라판이 파골세포 형성 억제에 대한 효과를 측정하였다. 설포라판은 대식세포인 RAW 264.7 세포에서 파골세포 특이 마커 유전자인 tartrate-resistant acid phosphatase(TRAP), Cathepsin K, matrix metalloproteinase 9(MMP-9), calcitonin receptor을 저해하였으며, TRAP, MMP-9, tumor necrosis factor receptor-associated factor 6(TRAF6)와 전사인자인 nuclease factor of activated T cells(NFATc1)의 단백질 발현과 RANKL로 자극하였을 때 전자인사인 nuclear factor kappaB(NF-kappaB)의 전사활성도 억제 하였다. 이와 같은 결과로 설포라판이 NF-kappaB의 전사활성 억제뿐만 아니라, 파골세포형성인자(TRAP, cathepsin K, MMP-9, calcitonin, NFATc1)와 NFATc1의 발현을 억제시키는 효과가 있음을 확인하였다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.197-203
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• 2023

제주상사화(Lycoris chejuensis)는 우리나라에 자생하는 특산식물로 관상용으로의 가치가 높을 뿐만 아니라 다양한 약리성분을 함유하고 있어 약용적으로도 가치가 높은 식물이다. 그러나 종자번식이 이루어지지 않고 구근을 통해 번식이 이루어지며 소수의 개체가 자생하므로 산업적으로 활용하기에 어려움이 따른다. 따라서 본 연구는 식물의 종 특성을 벗어나 환경에 관계없이 대량화 할 수 있는 제주상사화 callus (LC)를 사용하였으며 미생물의 효소 작용을 이용하여 분자의 구조 수정을 유도하는 친환경 생물전환 기법인 biorenovation을 적용하여 활성을 증진시키고자 하였다. 이에 본 연구에서는 biorenovation 된 제주상사화 callus 추출물(LCB)의 항염증 활성을 마우스 대식세포인 RAW 264.7 세포에서 평가하였으며 앞서 LC 및 LCB가 세포의 생존에 미치는 영향을 조사한 결과 25, 50, 100 ㎍/mL 농도에서 LCB는 LC의 높은 세포독성을 감소하였음을 확인하였다. 또한 LCB는 세포 독성이 나타나지 않는 농도에서 nitric oxide 및 prostaglandin E₂를 효과적으로 억제하였으며 이들의 합성 효소인 inducible NO synthase, cyclooxygenase-2의 발현을 유의하게 억제함으로써 NO 및 PG E₂를 효과적으로 하향 조절함을 입증하였다. 뿐만 아니라 pro-inflammatory cytokines인 tumor necrosis factor-α와 interleukin-1β, interleukin-6의 발현 또한 유의하게 억제하는 것으로 확인되었다. 이러한 결과를 통해 LCB가 다양한 염증 인자를 표적으로 하는 항염증 소재로 적용될 수 있음을 제안한다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.213-220
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• 2023

본 연구는 비자나무(Torreya nucifera (L.) Siebold & Zucc, TN)와 생물전환 된 비자나무 추출물(TNB)의 항염증 효과를 평가하기 위해 수행되었으며 이를 위해 C. acnes에 의해 유도된 RAW264.7 염증 모델에서 염증인자의 발현을 조사하였다. 실험 결과, TNB는 50, 100, 200 ㎍/mL 농도에서 TN의 높은 세포독성을 개선하였으며 nitric oxide (NO)와 NO 합성 효소인 inducible NO synthase (iNOS) 및 prostaglandin의 합성 효소인 cyclooxygenase-2 (COX-2)의 발현을 유의하게 억제하였다. 또한 TNB는 염증성 사이토카인인 tumor necrosis factor-α (TNF-α) 및 interleukin (IL)-1β, IL-6, IL-8의 발현을 유의하게 억제하였으며 특히 IL-6, IL-8의 경우 가장 고 농도인 200 ㎍/mL에서 정상세포 수준으로 감소하였다. 이후 진행된 western blot에서 인산화 된 IκB-α 및 NF-κB의 발현이 농도의존적으로 억제됨을 확인하였으며 인산화가 억제되면서 degradation이 감소하여 TNB처리 농도가 높아짐에 따라 IκB-α의 농도가 증가하는 경향을 보였다. 결론적으로, TNB는 NF-κB신호 전달 경로를 차단함으로써 다양한 염증 매개 인자의 발현을 효과적으로 하향 조절할 수 있으며 이를 통해 항 염증 활성을 유도하는 것으로 확인된다. 이러한 결과를 근거로 TNB가 C. acnes에 의해 유발된 염증성질환의 치료에 효과적인 천연물 소재로 적용될 수 있음을 제안한다.