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Validation of Neurotensin Receptor 1 as a Therapeutic Target for Gastric Cancer
Akter, Hafeza,Yoon, Jung Hwan,Yoo, Young Sook,Kang, Min-Jung Korean Society for Molecular and Cellular Biology 2018 Molecules and cells Vol.41 No.6
Gastric cancer is the fifth most common type of malignancy worldwide, and the survival rate of patients with advanced-stage gastric cancer is low, even after receiving chemotherapy. Here, we validated neurotensin receptor 1 (NTSR1) as a potential therapeutic target in gastric cancer. We compared NTSR1 expression levels in sixty different gastric cancer-tissue samples and cells, as well as in other cancer cells (lung, breast, pancreatic, and colon), by assessing NTSR1 expression via semi-quantitative real-time reverse transcription polymerase chain reaction, immunocytochemistry and western blot. Following neurotensin (NT) treatment, we analyzed the expression and activity of matrix metalloproteinase-9 (MMP-9) and further determined the effects on cell migration and invasion via wound-healing and transwell assays. Our results revealed that NTSR1 mRNA levels were higher in gastric cancer tissues than non-cancerous tissues. Both of NTSR1 mRNA levels and expression were higher in gastric cancer cell lines relative to levels observed in other cancer-cell lines. Moreover, NT treatment induced MMP-9 expression and activity in all cancer cell lines, which was significantly decreased following treatment with the NTSR1 antagonist SR48692 or small-interfering RNA targeting NTSR1. Furthermore, NT-mediated metastases was confirmed by observing epithelial-mesenchymal transition markers SNAIL and E-cadherin in gastric cancer cells. NT-mediated invasion and migration of gastric cancer cells were reduced by NTSR1 depletion through the Erk signaling. These findings strongly suggested that NTR1 constitutes a potential therapeutic target for the inhibition of gastric cancer invasion and metastasis.
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Akter, Hafeza,Park, Min,Kwon, Oh-Seung,Song, Eun Joo,Park, Won-Sang,Kang, Min-Jung Saikon Pub. Co 2015 TUMOR BIOLOGY Vol.36 No.8
<P>Neurotensin (NT) is distributed throughout the brain and gastrointestinal tract. Although the relationship between NT and matrix metalloproteinase-9 (MMP-9) activity in gastric cancer has not been reported, the elevation of MMP-9 and NT is reported in the breast, lung, prostate, and gastric cancer. The aim of our study is to investigate the relationship between NT and MMP-9 activity and the underlying signaling mechanism in gastric cancer cell lines. Commercial ELISA kits were used for estimation of NT and MMP-9 expression, and fluorescence resonance energy transfer (FRET) assay was used for measurement of MMP-9 activity. Cell migration and invasion were determined by wound healing and transwell assay. The expression of signaling proteins was measured by Western blotting. Our study reveals a positive correlation between increased plasma NT and MMP-9 activity in both of patient's serum and gastric cancer cell lines. A dose-dependent elevation of MMP-9 activity was observed by NT treatment in gastric cancer cells (MKN-1 and MKN-45) compared to untreated gastric cancer and normal epithelial cell (HFE-145). Moreover, NT-mediated migration and invasion were observed in gastric cancer cells unlike in normal cell. The signaling mechanism of NT in gastric cancer cells was confirmed in protein kinase C (PKC), extracellular-signal regulated kinase (ERK), and phosphatidylinositol 3-kinase (PI3K) pathway. In addition, pretreatment of gastric cancer cells with NTR1 inhibitor SR48692 was shown to significantly inhibit the NT-mediated MMP-9 activity, cell invasion, and migration. Our finding illustrated NTR1 could be a possible therapeutic target for gastric cancer.</P>
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자성나노입자 주입에 의한 혈청의 인터페론-감마 변화 특성
이한승,전찬호,HASAN MAHBUB,최종구,AKTERHAFEZA,배예빈,이상석 한국물리학회 2022 새물리 Vol.72 No.9
Magnetic nanoparticles are widely used for treatment and diagnosis in the medical field. Given that their position is easily manipulated using a magnetic field, they can be developed as targeted immunotherapeutic agents. This study aims to determine whether dextran coat on 40 nm Fe3O4 magnetic nanoparticles is medically toxic. The interferon-gamma (IFN-γ) concentration is measured using a sandwich ELISA method. IFN-γ is a potent inflammatory cytokine that increases with toxic immune responses. If magnetic nanoparticles are toxic, then an inflammatory reaction occurs when they are injected into the body through the tail vein of a mouse, resulting in a change in IFN-γ concentration in blood serum. However, when dextran-coated magnetic nanoparticles are injected in vivo, the IFN-γ concentration in the mouse serum does not increase, so an inflammatory reaction does not occur. Therefore, the dextran-coated magnetic nanoparticles are biocompatible and suitable for antibody-conjugated targeted immunotherapeutic drug development. 치료와 진단의 목적으로 의료분야에 많이 사용되고 있는 자성나노입자는 자기장에 의해 위치조작이 가능한 장점이 있어 표적 면역 치료제로 개발될 수 있다. 40 nm 크기의 자철석 Fe3O4 에 덱스트란이 코팅된 자성나노입자를 의학적으로 독성 여부를 확인하기 위해 샌드위치 ELISA 방법을 이용하여 인터페론-감마 (IFN-γ)의 농도를 측정하게 된다. 자성나노입자가 독성이 있다면 마우스 꼬리 정맥을 통해 체내 주입되었을 때 염증 반응이 일어나 혈액의 혈청 내 IFN-γ 농도의 변화를 나타내게 된다. 면역 반응이 일어나면 사이토카인이 분비되기 때문에 IFN-γ가 증가하게 된다. 덱스트란-자성나노입자를 생체 내에 주입하였을 때 마우스의 혈청 내의 IFN-γ가 증가하지 않았기에 염증 반응이 일어나지 않아서 독성이 없는 생체적합성 물질로 면역치료에 적합한 단일클론항체를 결합한 자성나노입자를 의약학적으로 사용이 가능할 수 있다.
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유도결합 플라즈마 질량분석법을 이용한 마우스 비장 내 Fe 입자 분포 연구
임예은,이정인,HASAN MAHBUB,최종구,AKTERHAFEZA,이상석 한국물리학회 2022 새물리 Vol.72 No.9
Magnetic nanoparticles (MNPs) are biocompatible materials. One example is iron (Fe), one of 14 trace elements constituting living things. MNPs are used in various bioapplications comprising drug delivery and thermotherapy. After anesthesia, mice were injected intravenously (tail vein) with either 50 μL of PBS (control group) or MNPs (treatment group) and maintained for 20 days. The MNPs were administered at a concentration of 2 μg/μL. The mice were sacrificed on days 0, 5, 10, 15, and 20 post-MNP injections. After anatomical examination, the organs were collected and wet weights were measured. The spleen, which stores immune cells and recycles red blood cells, was preprocessed by acid digestion. After a spleen sample in a homogeneous state was prepared in a 50 mL tube to a final volume of 20 mL, the level of inorganic Fe elements in the spleen was detected through ICP-MS. Furthermore, the intensity and concentration of Fe in the spleen of control mice and MNP-treated mice were identified. The intravenously injected dextran-MNPs showed the highest accumulation rate in the spleen on day 15. Therefore, the optimal period for suppressing the overactivation of T cells in the spleen is 15 days. 생체 적합성 재료로 사용되고 있는 자성나노입자는 생명체를 구성하는 14가지 미량 원소들 중 하나로서 약물전달, 발열요법 등 다양한 바이오 응용분야에 활용되고 있다. 마취가 된 마우스 꼬리 정맥에 1마리는 PBS만을 주입하고 나머지 4마리는 2 μg/μL 농도의 자성나노입자를 각각 주입하고 복부를 개복하여 총 7가지의 장기를 해부함과 동시에 중량 측정을 진행한다. 채취한 장기 중 T세포와 적혈구를 생성하는 비장만 분리하여 시료의 전 처리 과정을 수행한다. 50 mL 튜브에 균질액 상태인 비장 샘플을 최종 부피 20 mL로 만든 후 ICP-MS를 통해 시료를 구성하고 있는 성분 중 비장 내 Fe 무기원소의 수치를 검출한다. 대조군 마우스부터 20일차 마우스까지의 비장 내 Fe 강도와 비장 내 Fe 농도를 분석한다. 정맥으로 주입된 덱스트란-자성나노입자는 15일차에 비장에서 가장 높은 축적률을 보인다. 따라서 비장 내 T세포 과잉 활성화를 억제하기 위한 최적의 기간은 15일이다.
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