• BMB Reports
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• 제51권12호
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• pp.642-647
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• 2018

Ubiquitin-conjugating enzyme E2S (UBE2S), a family of E2 protein in the ubiquitination process, is involved in development of various cancers. However, its role in lung adenocarcinoma, has not been well elucidated. In this report, we attempted to investigate expression and function of UBE2S in lung adenocarcinoma. Up-regulation of UBE2S at mRNA, and protein level, was observed in human cancer tissues and lung adenocarcinoma cells. Higher UBE2S expression correlated with poorer prognosis of lung adenocarcinoma patients. UBE2S expression was efficiently suppressed by lentivirus-mediated shRNA strategy in A549 cells, and UBE2S silencing led to reduced cell proliferation, colony formation, and enhanced apoptosis. Inverse results were observed, in UBE2S over-expressed H1299 cells. Microarray analysis indicated that a large number of genes were regulated by UBE2S, and p53 signaling pathway may be critical, to the role of UBE2S in cancer development. Together, UBE2S could be a potential target for lung adenocarcinoma.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8247-8252
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• 2016

Background: The ubiquitin-proteasome system (UPS) degrades a variety of proteins which attach to specific signals. The ubiquitination pathway facilitates degradation of damaged proteins and regulates growth and stress responses. This pathway is altered in various cancers, including acute lymphoblastic leukemia, head and neck squamous cell carcinoma and breast cancer. Recently it has been reported that expression of newly characterized human genes, UBE2Q1 and UBE2Q2, putative members of ubiquitin-conjugating enzyme family (E2), has been also changed in colorectal cancer. Epigenetics is one of the fastest-growing areas of science and nowadays has become a central issue in biological studies of diseases. According to the lack of information about the role of epigenetic changes on gene expression profiling of UBE2Q1 and UBE2Q2, and the presence of CpG islands in the promoter of these two human genes, we decided to evaluate the promoter methylation status of these genes as a first step. Materials and Methods: The promoter methylation status of UBE2Q1 and UBE2Q2 was studied by methylation-specific PCR (MSP) in tumor samples of 60 colorectal cancer patients compared to adjacent normal tissues and 20 non-malignant controls. The frequency of the methylation for each gene was analyzed by chi-square method. Results: MSP results revealed that UBE2Q2 gene promoter were more unmethylated, while a higher level of methylated allele was observed for UBE2Q1 in tumor tissues compared to the adjacent normal tissues and the non malignant controls. Conclusions: UBE2Q1 and UBE2Q2 genes show different methylation profiles in CRC cases.

• Journal of Microbiology and Biotechnology
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• 제32권6호
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• pp.740-748
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• 2022

As circ_UBE2D2 has been confirmed to have targeted binding sites with multiple miRNAs involved in septic acute kidney injury (SAKI), efforts in this study are directed to unveiling the specific role and relevant mechanism of circ_UBE2D2 in SAKI. HK-2 cells were treated with lipopolysaccharide (LPS) to construct SAKI model in vitro. After sh-circ_UBE2D2 was transfected into cells, the transfection efficiency was detected by qRT-PCR, cell viability and apoptosis were determined by MTT assay and flow cytometry, and expressions of Bcl-2, Bax and Cleaved-caspase 3 were quantified by western blot. Target genes associated with circ_UBE2D2 were predicted using bioinformatics analysis. After the establishment of SAKI rat model, HE staining and TUNEL staining were exploited to observe the effect of circ_UBE2D2 on tissue damage and cell apoptosis. The expression of circ_UBE2D2 was overtly elevated in LPS-induced HK-2 cells. Sh-circ_UBE2D2 can offset the inhibition of cell viability and the promotion of cell apoptosis induced by LPS. Circ_UBE2D2 and miR-370-3p as well as miR-370-3p and NR4A3 have targeted binding sites. MiR-370-3p inhibitor reversed the promoting effect of circ_UB2D2 silencing on viability of LPS-treated cells, but shNR4A3 neutralized the above inhibitory effect of miR-370-3p inhibitor. MiR-370-3p inhibitor weakened the down-regulation of NR4A3, Bax and Cleaved caspase-3 and the up-regulation of Bcl-2 induced by circ_UB2D2 silencing, but these trends were reversed by shNR4A3. In addition, sh-circ_UBE2D2 could alleviate the damage of rat kidney tissue. Circ_UBE2D2 mitigates the progression of SAKI in rats by targeting miR-370-3p/NR4A3 axis.

• 생명과학회지
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• 제33권6호
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• pp.523-529
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• 2023

유비퀴틴-프로테아좀 시스템은 E1-E2-E3 효소의 작용으로 단백질 안정성을 조절하며 이를 통해 진핵 세포 내 광범위한 과정을 조절한다. 특히 DNA 수리, 세포 주기, 전이, 혈관형성 및 사멸과 같은 종양의 생장 과정에서 주요한 역할을 하는데 이 과정에서 유비퀴틴 접합 효소인 UBE2는 활성화된 유비퀴틴을 타깃 단백질에 이동시켜주는 중간 매개체 역할을 한다. UBE2는 인간에게서 40개가 존재하며 이는 촉매 도메인의 확장 유무에 따라 4개의 그룹으로 분류된다. 최근 UBE2의 타깃 단백질의 특정 위치를 인식하는 기질 특이성에 대한 연구가 증가하는 추세이다. 특히 암에서 발현이 높은 UBE2는 암 환자의 나쁜 예후와 상관관계가 있어 종양 형성에서 UBE2의 중요성이 강조되고 있다. 본 총설에서는 암에서 UBE2의 역할에 대한 최신 연구 결과 및 동향에 대해 기술하였다. 또한 UBE2에 관한 기초 지식 및 분자적 메커니즘을 제공함으로써 궁극적으로는 UBE2가 종양 치료의 새로운 타깃이 될 수 있음을 시사한다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3005-3009
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• 2014

The aims of this study were to investigate the influence of ubiquitin- conjugating enzyme E2C (UBE2C) on biological behavior of lung cancer cells. Using MTT, flow cytometry and invasion assays, we detected UBE2C expression and evaluated its biological properties in these cells, including effects on proliferation, the cell cycle profile and invasive capability. Compared with control cells, the UBE2C transfected cells demonstrated increased cellular proliferation (p<0.05). UBE2C transfected cells also had a lower percentage in G1 phase and a higher percentage in S phase (p<0.05). Importantly, the UBE2C transfected cells had a notable enhancement of cell numbers penetrating the basement membrane compared with the control group (p<0.05). Ectopic up-regulation UBE2C promoted the growth of lung cancer cells in vivo. Furthermore, we found UBE2C increased the expression of cyclin D1 and MMP-2. These results show UBE2C may represent a potential therapeutic target for lung cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3723-3727
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• 2015

The p53 tumor suppressor protein is a principal mediator of growth arrest, senescence, and apoptosis in response to a broad array of cellular damage. p53 is a substrate for the ubiquitin-proteasome system, however, the ubiquitin-conjugating enzymes (E2s) involved in p53 ubiquitination have not been well studied. UBE2Q1 is a novel E2 ubiquitin conjugating enzyme gene. Here, we investigated the effect of UBE2Q1 overexpression on the level of p53 in the MDA-MB-468 breast cancer cell line as well as the interaction between UBE2Q1 and p53. By using a lipofection method, the p53 mutated breast cancer cell line, MDA-MB-468, was transfected with the vector pCMV6-AN-GFP, containing UBE2Q1 ORF. Western blot analysis was employed to verify the overexpression of UBE2Q1 in MDA-MB-468 cells and to evaluate the expression level of p53 before and after cell transfection. Immunoprecipitation and GST pull-down protocols were used to investigate the binding of UBE2Q1 to p53. We established MDA-MB-468 cells that transiently expressed a GFP fusion proteins containing UBE2Q1 (GFP-UBE2Q1). Western blot analysis revealed that levels of p53 were markedly lower in UBE2Q1 transfected MDA-MB-468 cells as compared with control MDA-MB-468 cells. Both in vivo and in vitro data showed that UBE2Q1 co-precipitated with p53 protein. Our data for the first time showed that overexpression of UBE2Q1can lead to the repression of p53 in MDA-MB-468 cells. This repression of p53 may be due to its UBE2Q1 mediated ubiquitination and subsequent proteasome degradation, a process that may involve direct interaction of UBE2Q1with p53.

• 한국발생생물학회지:발생과생식
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• 제12권3호
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• pp.289-296
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• 2008

BPES($\underline{B}$lepharophimosis/$\underline{P}$tosis/$\underline{E}$picanthus inversus $\underline{S}$yndrome)는 FOXL2 유전자의 돌연변이에 의해 유발되는 상염색체 우성질환이다. 눈꺼풀이 갈라지거나 쳐지고 넓은 미간이 나타나는 특징이 있으며, 여성의 조기 난소 부전증(premature ovarian failure, POF)을 일으켜 불임을 유발한다. FOXL2는 forkhead family에 속하는 전사인자로서 FOXL2가 결여된 난소에서는 granulosa cell의 분화가 진행되지 않아 난포 성숙과정의 멈춤과 난자의 폐쇄증을 유발한다. FOXL2를 bait로 하여 rat의 난소 cDNA 라이브러리의 yeast two-hybrid screening을 시행하여 FOXL2 단백질과 상호작용을 하는 small ubiquitin-related modifier(SUMO)-conjugating E2 효소인 UBE2I 단백질을 찾았다. UBC9이라고도 알려진 UBE2I 단백질은 SUMO 변형 과정을 위한 필수적인 단백질이다. Sumoylation은 수 많은 전사인자의 전사능력의 조절을 포함하여 다양한 신호전달체계에 관여하는 번역 후 변형 과정이다. 본 연구에서 인간세포인 293T 내에서 면역침전반응 실험을 통해 FOXL2와 UBE2I의 단백질-단백질간의 상호작용을 확인하고, FOXL2의 돌연변이형을 제작하여 yeast two-hybrid system을 이용해 UBE2I와 결합에 필요한 FOXL2의 부분을 규명하였다. 따라서, FOX2에 상호작용하는 UBE2I의 규명은 sumoylation에 의한 FOXL2의 새로운 조절 메커니즘을 시사한다.

• Clinical Endoscopy
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• 제51권6호
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• pp.596-599
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• 2018

An 89-year-old man was referred to our hospital for treatment of hepatolithiasis causing recurrent cholangitis. He had undergone a prior Whipple procedure. Computed tomography demonstrated left-sided hepatolithiasis. First, we conducted peroral direct cholangioscopy (PDCS) using an ultraslim endoscope. Although PDCS was successfully conducted, it was unsuccessful in removing all the stones. The stones located in the B2 segment were difficult to remove because the endoscope could not be inserted deeply into this segment due to the small size of the intrahepatic bile duct. Next, we substituted the endoscope with an upper gastrointestinal endoscope. After positioning the endoscope, the SpyGlass digital system (SPY-DS) was successfully inserted deep into the B2 segment. Upon visualizing the residual stones, we conducted SPY-DS-guided electrohydraulic lithotripsy. The stones were disintegrated and completely removed. In cases of PDCS failure, a treatment strategy using the SPY-DS can be considered for patients with hepatolithiasis after a Whipple procedure.

• Toxicological Research
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• 제32권1호
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• pp.73-80
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• 2016

Chronic exposure to cadmium (Cd) is known to adversely affect renal function. Our previous studies indicated that Cd induces p53-dependent apoptosis by inhibiting gene expression of the ubiquitin-conjugating enzyme (Ube) 2d family in both human and rat proximal tubular cells. In this study, the effects of Cd on protein expression of p53 and apoptotic signals in the kidney and liver of mice exposed to Cd for 12 months were examined, as well as the effects of Cd on p53 protein levels and gene expression of the Ube2d family in various cell lines. Results showed that in the kidney of mice exposed to 300 ppm Cd for 12 months, there was overaccumulation of p53 proteins in addition to the induction of apoptosis, which was triggered specifically in the proximal tubules. Interestingly, the site of apoptosis was the same as that of p53 accumulation in the proximal tubules. In the liver of mice chronically exposed to Cd, gene expression of the Ube2d family tended to be slightly decreased, together with slight apoptosis without the accumulation of p53 protein. In rat small intestine epithelial (IEC-6) cells, Cd decreased not only the p53 protein level but also gene expression of Ube2d1, Ube2d2 and Ube2d4. In human brain microvascular endothelial cells (HBMECs), Cd did not suppress gene expression of the Ube2d family, but increased the p53 protein level. In human brain astrocytes (HBASTs), Cd only increased gene expression of UBE2D3. These results suggest that Cd-induced apoptosis through p53 protein is associated with renal toxicity but not hepatic toxicity, and the modification of p53 protein by Cd may vary depending on cell type.

• 한국전자현미경학회:학술대회논문집
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• 한국현미경학회 2005년도 제36차 춘계학술대회 및 제3회 HVEM 이용자 워크샵
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• pp.141-144
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• 2005