• IMMUNE NETWORK
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• 제21권4호
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• pp.30.1-30.12
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• 2021

High expression of mitofusin-2 (MFN2), a mitochondrial fusion protein, has been frequently associated with poor prognosis of patients with cervical cancer. Here, we aimed to identify the function of MFN2 in cervical cancer to understand its influence on disease prognosis. To this end, from cervical adenocarcinoma, we performed an MTT assay and quantitative RT-PCR (qRT-PCR) analysis to assess the effects of MFN2 on the proliferation and of HeLa cells. Then, colony-formation ability and tumorigenesis were evaluated using a tumor xenograft mouse model. The migration ability related to MFN2 was also measured using a wound healing assay. Consequently, epithelial-mesenchymal transition (EMT) of MFN2-knockdowned HeLa cells originating from adenocarcinoma. markers related to MFN2 were assessed by qRT-PCR. Clinical data were analyzed using cBioPortal and The Cancer Genome Atlas. We found that MFN2 knockdown reduced the proliferation, colony formation ability, migration, and in vivo tumorigenesis of HeLa cells. Primarily, migration of MFN2-knockdowned HeLa cells decreased through the suppression of EMT. Thus, we concluded that MFN2 facilitates cancer progression and in vivo tumorigenesis in HeLa cells. These findings suggest that MFN2 could be a novel target to regulate the EMT program and tumorigenic potential in HeLa cells and might serve as a therapeutic target for cervical cancer. Taken together, this study is expected to contribute to the treatment of patients with cervical cancer.

• Biomolecules & Therapeutics
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• 제32권1호
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• pp.115-122
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• 2024

Heat shock protein (HSP) 90 is expressed in most living organisms, and several client proteins of HSP90 are necessary for cancer cell survival and growth. Previously, we found that HSP90 was cleaved by histone deacetylase (HDAC) inhibitors and proteasome inhibitors, and the cleavage of HSP90 contributes to their cytotoxicity in K562 leukemia cells. In this study, we first established mouse xenograft models with K562 cells expressing the wild-type or cleavage-resistant mutant HSP90β and found that the suppression of tumor growth by the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) was interrupted by the mutation inhibiting the HSP90 cleavage in vivo. Next, we investigated the possible function of thioredoxin interacting protein (TXNIP) in the HSP90 cleavage induced by SAHA. TXNIP is a negative regulator for thioredoxin, an antioxidant protein. SAHA transcriptionally induced the expression of TXNIP in K562 cells. HSP90 cleavage was induced by SAHA also in the thymocytes of normal mice and suppressed by an anti-oxidant and pan-caspase inhibitor. When the thymocytes from the TXNIP knockout mice and their wild-type littermate control mice were treated with SAHA, the HSP90 cleavage was detected in the thymocytes of the littermate controls but suppressed in those of the TXNIP knockout mice suggesting the requirement of TXNIP for HSP90 cleavage. We additionally found that HSP90 cleavage was induced by actinomycin D, β-mercaptoethanol, and p38 MAPK inhibitor PD169316 suggesting its prevalence. Taken together, we suggest that HSP90 cleavage occurs also in vivo and contributes to the anti-cancer activity of various drugs in a TXNIP-dependent manner.

• Journal of Microbiology and Biotechnology
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• 제34권4호
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• pp.774-782
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• 2024

This study aimed to elucidate the anti-colon cancer mechanism of ginsenoside Rg1 in vitro and in vivo. Cell viability rate was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium assay. The inhibitory effect of ginsenoside Rg1 against CT26 cell proliferation gradually increased with increasing concentration. The in vivo experiments also demonstrated an antitumor effect. The monodansylcadaverine (MDC), transmission electron microscopy (TEM), and expression of autophagy marker proteins confirmed that ginsenoside Rg1 induced autophagy in vitro. Ginsenoside Rg1 induced autophagy death of CT26 cells, but this effect could be diminished by autophagy inhibitor (3-methyladenine, 3-MA). Additionally, in a xenograft model, immunohistochemical analysis of tumor tissues showed that the LC3 and Beclin-1 proteins were highly expressed in the tumors from the ginsenoside Rg1-treated nude mice, confirming that ginsenoside Rg1 also induced autophagy in vivo. Furthermoer, both in vivo and in vitro, the protein expressions of p-Akt, p-mTOR, and p-p70S6K were inhibited by ginsenoside Rg1, which was verified by Akt inhibitors. These results indicated that the mechanism of ginsenoside Rg1 against colon cancer was associated with autophagy through inhibition of the Akt/mTOR/p70S6K signaling pathway.

• Molecules and Cells
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• 제44권7호
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• pp.468-480
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• 2021

Ubiquitin D (UBD) is highly upregulated in many cancers, and plays a pivotal role in the pathophysiological processes of cancers. However, its roles and underlying mechanisms in oral squamous cell carcinoma (OSCC) are still unclear. In the present study, we investigated the role of UBD in patients with OSCC. Quantitative real-time polymerase chain reaction and Western blot were used to measure the expression of UBD in OSCC tissues. Immunohistochemistry assay was used to detect the differential expressions of UBD in 244 OSCC patients and 32 cases of normal oral mucosae. In addition, CCK-8, colony formation, wound healing and Transwell assays were performed to evaluate the effect of UBD on the cell proliferation, migration, and invasion in OSCC. Furthermore, a xenograft tumor model was established to verify the role of UBD on tumor formation in vivo. We found that UBD was upregulated in human OSCC tissues and cell lines and was associated with clinical and pathological features of patients. Moreover, the overexpression of UBD promoted the proliferation, migration and invasion of OSCC cells; however, the knockdown of UBD exerted the opposite effects. In this study, our results also suggested that UBD promoted OSCC progression through NF-κB signaling. Our findings indicated that UBD played a critical role in OSCC and may serve as a prognostic biomarker and potential therapeutic target for OSCC treatment.

• 한국미생물·생명공학회지
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• 제47권1호
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• pp.164-171
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• 2019

기존에는 방사선 요법은 효과에 비해 소화기 점막 궤양 등 부작용이 많다. 본 연구는 방사선의 작용을 모사하는 방사선 모사 미생물 유래 리보솜 스트레스 반응을 이용하여 방사선 치료의 한계를 대체할 기전적 방법을 모색하고자 한다. 암 치료의 평가를 위해서 대장암의 비균질적인 세포구성, 종양줄기세포 및 주위 미세환경 및 배양 기질 등의 상호작용을 고려할 수 있는 소화기암 스페로이드를 이용하였다. 대조군 대장암 스페로이드에 비해서 리보솜 스트레스하에서는 스페로이드 구조가 상대적으로 큰 군집을 형성하였다. 하지만 이는 구조 자체가 매우 섬긴 세포간 구성을 가진 스페로이드였으며 스페로이드 형성과정의 크기 수축은 대조군에 비해 매우 느리게 나타났다. 대조군은 강하게 뭉친 소규모 클러스트의 집합체가 최종적으로 스페로이드를 형성하여 물리적 손상을 받아도 단단한 소규모 클러스트는 유지되나 리보솜 스트레스 하에서는 물리적 손상에 의해 형태가 파손 후에는 스페로이드 형성이 거의 일어나지 않았다. 기전적으로 리보솜 스트레스 하에서 암세포의 군집하는 이동속도는 매우 느렸으며, 뭉치더라도 세포-세포간 접합부위가 상대적으로 적었다. 이 대장암 스페로이드를 이종 및 동종 이식을 통해 동물에서 증식 시 종양 조직 형성이 매우 억제되었으며, 형성이 되어도 세포-세포간 접합에 핵심단백질인 E-cadherin의 발현이 매우 감소 됨을 알 수 있었다. 결론적으로 방사선 모사 미생물 유래 리보솜 스트레스는 종양 스페로이드 세포 이동 및 접합을 저해하여 3차원 구조 형성 결함을 유발하였으며, 향후 방사선을 대체하여 약물적으로 방사선 항암효과를 구현하는 기반을 제공한다.

• 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.

• 대한방사성의약품학회지
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• 제7권2호
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• pp.105-111
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• 2021

⁸⁹Zr is a positron-emitting radioisotope, which has known as well-suited radioisotope for use in a monoclonal antibody-based imaging agent for immuno-PET. The purpose of this study was to quantitatively evaluate the diagnostic ability of general trastuzumab and thio-trastuzumab as HER2 positive receptors based on Df hexadentate iron chelator. Desferrioxamine-p-SCN (Df-Bz-NCS) and desferroixamine-maleimide (Df-Mal) were purchased from Macrocyclics (Dallas, TX, USA). The trastuzumab was purchased from Roche (Schweiz), and thio-trastuzumab was obtained from professor Hyo-Jeong Hong group (Kangwon National University). The radioisotope ⁸⁹Zr was produced by domestic purification system and KIRAMS using medical cyclotron (50 MeV, Scantronix). The conjugates of Df-trastuzumab and Df-thio-trastuzumab were prepared with Df-Bz-NCS and Df-Mal under basic aqueous solution (pH 8-9) at room temperature, respectively. The conjugates purified by PD-10 column were mixed with dried ⁸⁹Zr chloride. ⁸⁹Zr-labeled conjugates were purified and concentrated by Amicon ultra centrifugal filter. The preparation step and time of ⁸⁹Zr-labeled conjugates was shorted as 4 steps within 2 hours. ⁸⁹Zr-labeled conjugates showed the highly radiochemical purity of over 98%, and were very stable until 7 days by the analysis of radio-ITLC method. Each radio-labeled conjugates were also exhibited the highly stability in both PBS buffer and mouse serum. Immuno-PET imaging of ⁸⁹Zr-labeled conjugates in mice bearing gastric cancer xenograft tumors with HER2 expression showed high tumor uptake in the NCI-N87 HER2-expressing. However, ⁸⁹Zr-Df-Mal-thio-trastuzumab showed a relatively lower tumor-to-background ratio than ⁸⁹Zr-Df-Bz-trastuzumab, as well as whole-body distribution. In the results, ⁸⁹Zr-Df-Bz-trastuzumab was evaluated to have a relatively higher HER2 diagnostic ability than ⁸⁹Zr-Df-Mal-thio-trastuzumab.

• 생명과학회지
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• 제34권5호
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• pp.287-295
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• 2024

간세포암종은 전 세계적으로 암 관련 사망의 두 번째 주요 원인으로 새로운 치료 전략을 필요로 한다. 간세포암종 환자를 치료에 사용되는 화학요법제는 독성이 있고 심각한 부작용이 있는 것으로 알려졌다. 본 연구는 건강한 간세포에서 세포독성을 일으키지 않으면서 종양세포를 표적으로 하는 부작용을 감소시키는 항암제의 효능을 조사하였다. Berberine은 이소퀴놀린 알칼로이드의 식물 유래 화합물로 다양한 약리학적 특성으로 인해 암 치료의 잠재적 후보군으로 보고되고 있다. 간암 세포 생존율에 대한 berberine의 효과는 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide 분석을 사용하여 측정하였다. HCC 증식은 콜로니 형성 분석을 통해 실시하였다. 세포이동에 대한 berberine의 효과는 상처 치유 분석으로 실시하였다. Berberine은 HepG2 세포와 같은 간암 세포의 증식과 콜로니 형성 및 세포이동을 억제하였다. Berberine 처리는 Beclin-1 및 LC3-II 등의 자가포식 관련 유전자 발현과 단백질 발현을 증가시켰으며, Caspase-3 및 Caspase-9등의 세포자멸사의 mRNA 발현 및 활성을 증가시켰다. 또한, 세포유래 이종이식 동물실험에서 berberine 처리에 따른 종양의 크기와 무게가 감소함을 확인하였다. 본 연구 결과는 간세포암에 대한 베르베린의 효능을 조명하여 표적 및 맞춤형 치료의 기회를 제시할 수 있음을 시사한다.

• Ultrasonography
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• 제32권2호
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• pp.132-142
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• 2013

목적: 전립선암 (PC-3, LNCaP)의 이종이식쥐 모델에서 초음파 조영제를 이용하여 측정한 시간-신호강도 곡선을 통해 얻은 역학적 변수를 조직병리적 변수와 비교하고자 하였다. 대상 및 방법: 20마리의 누드마우스에 인간전립선암세포(15 PC-3, 5 LNCaP)를 다리에 주입하여 이종이식쥐 모델을 제작하였다. 이후 $500{\mu}L$의 2세대 초음파 조영제인 Sonovue를 후안와정맥을 통해 주입하였다. 관심영역은 종양의 전체를 포함할 수 있게 그린 후, 시간-신호강도 곡선을 얻고 이를 감마변량함수에 적합시켰다. 최고신호강도(A), 최고도달시간 (Tp), 최고유입률 (washin), 최고 유출률 (washout), 50초까지의 곡선하면적 ($AUC_{50}$), 유입기면적 ($AUC_{in}$), 유출기면적 ($AUC_{out}$) 등을 감마변량함수에서 도출하였다. VEGF와 CD31에 대한 면역조직화학염색법을 시행하였고, 종양부피, 시야당 VEGF 면적백분율, CD31 양성 미세혈관수 (MVD) 등을 계산한 후 이를 시간신호강도에서 얻은 역학적 변수와 상관성을 조사하였다. 결과: PC-3 와 LNCaP간 동적 및 조직병리학적 변수의 통계적 차이는 없었다. MVD는 A (r=0.625, p=0.003), washin (r=0.462, p=0.040), AUC (r=0.604, p=0.005), $AUC_{out}$ (r=0.587, p=0.007) 과 유의한 상관관계를 보였다. 또한 종양부피와 $AUC_{50}$ (r=0.481, p=0.032), washin(r=0.662, p=0.001), $AUC_{out}$ (r=0.547, p=0.012) 도 유의한 양의 상관관계를 보였으며, washout은 MVD (r=-0.454, p=0.044) 및 종양부피 (r=-0.464, p=0.039)에 모두 음의 상관관계를 보였다. 그러나 시야당 VEGF 면적백분율은 역학적 변수와 유의한 상관관계를 보이지 않았다. 결론: MVD는 다수의 역학적 변수와 상관관계를 보였다. 초음파 조영제를 이용한 초음파는 전립선암 동물모델에서 종양의 혈관성을 예측할 수 있는 가능성을 보였다.

• 생명과학회지
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• 제18권10호
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• pp.1395-1399
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• 2008

우리는 최근에 PRC1 프로모터가 유방암 유전자치료를 위하여 전사 표적이 된 유전자의 발현을 조절할 수 있는 후보 프로모터임을 보고하였다. 우리는 본 실험에서 PRC1이 폐암유전자 치료에서도 적용이 가능한지 조사하였다. 특정 프로모터가 루시퍼라제 유전자와 연결된 플라스미드를 이용한 형질전환 assay에서 PRC1 프로모터는 정상폐세포주에서는 활성을 보이지 않으나 폐암세포주에선 약 30 배의 활성을 보였다. 이는 이미 암특이적인 발현으로 알려진 BIRC5 (survivin) 프로모터와 유사한 결과였다. 또한, 바이러스 벡터를 이용한 실험에서 PRC1은 CMV 프로모터에 비해 아데노부속바이러스에서 약 75%, 아데노바이러스에서 약 66%의활성을 보였다. 이와 대조적으로, PRC1 프로모터를 함유한 이 들 두 종류의 바이러스는 정상 폐세포에서는 20%정도의 낮은 활성을 보였다. 흥미롭게도, 인간 폐종양세포를 이식한 생쥐모델을 사용한 결과에서는 PRC1 프로모터가 CMV 프로모터와 비슷한 생체 활성을 보였다. 종합하면, 이상의 결과는 PRC1이 폐암 유전자치료를 위한 전사표적 유전자의 발현을 위한 프로모터로 사용 가능함을 암시한다.