• Title/Summary/Keyword: mtDNA

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Alterations in Mitochondrial DNA Copy Numbers and Mitochondrial Oxidative Phosphorylation (OXPHOS) Protein Levels in Gastric Cancer Tissues and Cell Lines (위암 조직과 세포주에서 mDNA와 OXPHOS 단백질 분석)

  • Siregar, Adrian;Hah, Young-Sool;Moon, Dong Kyu;Woo, Dong Kyun
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1057-1065
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    • 2021
  • Alterations in mitochondrial DNA (mtDNA) copy numbers have been reported in patients with stomach cancer and suggested to play a role in gastric carcinogenesis or gastric cancer progression. However, changes in the levels of mitochondrial proteins or mtDNA-encoded oxidative phosphorylation (OXPHOS) proteins in gastric cancer remain unclear. In this study, we investigated mtDNA contents, mitochondrial protein levels, and mtDNA-encoded OXPHOS protein levels in gastric cancer tissues and cell lines. We correlated mtDNA copy numbers with clinicopathologic features of the gastric cancer samples used in this study and used quantitative PCR to analyze the mtDNA copy numbers of the gastric cancer tissues and cell lines. Western blot analysis was used for assessing the amounts of mitochondrial proteins and mtDNA-encoded OXPHOS proteins. Among the 27 gastric cancer samples, 22 showed a reduction in mtDNA copy numbers. The mtDNA content was increased in the other five samples relative to that in normal matched gastric tissues. Mitochondrial protein and OXPHOS protein levels were reduced in some gastric cancer tissues. However, mitochondrial protein and OXPHOS protein levels in gastric cancer cell lines were not always in line with their mtDNA contents. The mtDNA copy numbers were reduced in five gastric cancer cell lines tested in this study. In summary, this study reports a common reduction in mtDNA contents in gastric carcinoma tissues and cell lines, pointing to the possible involvement of mtDNA content alterations in tumorigenesis of the stomach.

restriction Site Polymorphism of mtDNA for differentiating Anopheles quadrimaculatus (Say) Sibling Species (미토콘드리아 DNA 제한효소 절단부위 변이에 의한 Anopheles quadrimaculatus (Say) 모기의 자매종 구별)

  • ;S.K. Narang
    • Korean journal of applied entomology
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    • v.29 no.2
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    • pp.132-135
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    • 1990
  • Three mitochondrial cDNA probes from Aedes albopictus were used to demonstrate restriction site polymorphism in mtDNA of three sibling species of Anopheles quadrimaculatus(Say). It was shown by DNA hybridization to have substantial sequence homology betwen the mtDNA of different genus. The proves reveled local restriction site variation between members of the Anopheles quadrimaculatus sibling species complex. Mitochondrical DNA (mtDNA), isolated from individual mosquitoes was digested by type II restriction enzymes and four enzymes were found to be useful for the purpose. Hind III alone could be used to obtain a diagnostic restriction pattern.

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Variation of Mitochondrial DNA in Striped Field Mice, Apodemus agrarius coreae Thomas(Mammalia, Rodentia), from the Korean Penisula (한반도산 등줄쥐 Apodemus agrarius corease Thomas(포유강, 설치목)의 미토콘드리아 DNA의 변이)

  • 고흥선;유상규;김상복;유병선
    • Animal Systematics, Evolution and Diversity
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    • v.9 no.2
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    • pp.171-179
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    • 1993
  • Thirty nine samples of striped field mice (Apodemus agrarius coreae Thomas) from eight localities in the Korean peninsula were used for the analyses of mitochondria1 DNA (mtDNA) fragment patterns resulted from the digestion with eight restriction enzymes. A total of 31 fragments were recognized and seven mtDNA clones were revealed: one clone consisted of 32 among 39 samples from eight localities (1 of 1 from Sogcho, 4 of 5 from Mt. Chiak, 3 of 3 from Mt. Weolak, 2 of 2 from Mt. Sogri, 2 of 2 from Mt. Deokyoo, 3 of 4 from Mt. Jiri, 2 of 4 from Haenam, and 15 of 18 from Cheongju). The nucleotide-sequence divergences (p) among seven mtDNA clones ranged from 0.2% to 2.3% and distinct subgroups were not resulted from the grouping of these clones. It is confirmed that striped field mcie from the Korean peninsula is a single subspecies of Apodemus agrarius (A. agrarius coreae) because they were not divided into separate subgroups in their mtDNA genotypes.

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Variation of Mitochondrial DNA Restriction Fragments within One Subspecies of Korean Field mice, Apodemus peninsulae peninsulae Thomas (Mammalia: Rodentia), from Korea (한국에서 서식하는 흰넓적다리붉은쥐 한 아종 Apodemus peninsulae peninsulae Thomas(포유강: 설치목)의 미토콘드리아 DNA 절단단편의 변이)

  • Hung Sun Koh
    • Animal Systematics, Evolution and Diversity
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    • v.11 no.2
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    • pp.291-299
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    • 1995
  • Sampels of Korean field mice (apodemus peninsulae peninsulae Thomas ) from six localities in Korea were used for the analyses of mitochondrial DNa (Mt DNA) fragment patterns resulted from the digestion with eight restriction enzymes. A total of 29 fragments were recognized and seven mtDNA clones were revealed. The nucleotide-sequence divergences (p) among the seven mtDNA clones ranged from 0.42% to 2.01%. Moreover, the seven clones were grouped into three major subgroups with the mean divergence value of 1.52% among them. One subgroup was composed of three clones of 18 sample from three localities (16, Cheongu: 1, Mt. Sobaek : 1, Mt. weolak) L another subgroup, three clones of eight samples from four localities (2, Cheongju ; 2 , Mt. Weolak ; 2, Mt. Gaya ; 2, haenam) ; and the last subgroup, one clone of two samples from Cheongju. Three subgroups were also distinct with one another in their mtDNA genotypes of Stu I and the former two subgroups differed from the last subgroup in their genotypes with Pvu II. Further analyses with additional samples from various localities in Korea appeared to be necessary in order to clarify the taxonomic status of the distinct mtDNA subgroups.

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Mitochondrial genome editing: strategies, challenges, and applications

  • Kayeong Lim
    • BMB Reports
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    • v.57 no.1
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    • pp.19-29
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    • 2024
  • Mitochondrial DNA (mtDNA), a multicopy genome found in mitochondria, is crucial for oxidative phosphorylation. Mutations in mtDNA can lead to severe mitochondrial dysfunction in tissues and organs with high energy demand. MtDNA mutations are closely associated with mitochondrial and age-related disease. To better understand the functional role of mtDNA and work toward developing therapeutics, it is essential to advance technology that is capable of manipulating the mitochondrial genome. This review discusses ongoing efforts in mitochondrial genome editing with mtDNA nucleases and base editors, including the tools, delivery strategies, and applications. Future advances in mitochondrial genome editing to address challenges regarding their efficiency and specificity can achieve the promise of therapeutic genome editing.

Alteration of mitochondrial DNA content modulates antioxidant enzyme expressions and oxidative stress in myoblasts

  • Min, Kyung-Ho;Lee, Wan
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.6
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    • pp.519-528
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    • 2019
  • Mitochondrial dysfunction is closely associated with reactive oxygen species (ROS) generation and oxidative stress in cells. On the other hand, modulation of the cellular antioxidant defense system by changes in the mitochondrial DNA (mtDNA) content is largely unknown. To determine the relationship between the cellular mtDNA content and defense system against oxidative stress, this study examined a set of myoblasts containing a depleted or reverted mtDNA content. A change in the cellular mtDNA content modulated the expression of antioxidant enzymes in myoblasts. In particular, the expression and activity of glutathione peroxidase (GPx) and catalase were inversely correlated with the mtDNA content in myoblasts. The depletion of mtDNA decreased both the reduced glutathione (GSH) and oxidized glutathione (GSSG) slightly, whereas the cellular redox status, as assessed by the GSH/GSSG ratio, was similar to that of the control. Interestingly, the steady-state level of the intracellular ROS, which depends on the reciprocal actions between ROS generation and detoxification, was reduced significantly and the lethality induced by $H_2O_2$ was alleviated by mtDNA depletion in myoblasts. Therefore, these results suggest that the ROS homeostasis and antioxidant enzymes are modulated by the cellular mtDNA content and that the increased expression and activity of GPx and catalase through the depletion of mtDNA are closely associated with an alleviation of the oxidative stress in myoblasts.

Mitochondrial DNA Levels in Blood and Tissue Samples from Breast Cancer Patients of Different Stages

  • Xia, Peng;Wang, Hui-Juan;Geng, Ting-Ting;Xun, Xiao-Jie;Zhou, Wen-Jing;Jin, Tian-Bo;Chen, Chao
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1339-1344
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    • 2014
  • Aims: Alterations in mitochondrial DNA (mtDNA) have been implicated in carcinogenesis and tumor progression. We here evaluated the diagnostic and prognostic potential of mtDNA as a biomarker for breast cancer. Methods: Using multiplex real-time polymerase chain reaction, nuclear DNA (nDNA) and mtDNA levels in serum, buffy coat, tumor, and tumor-adjacent tissue samples from 50 breast cancer patients were determined and assessed for associations with clinicopathological features. To evaluate mtDNA as a biomarker for distinguishing between the four sample types, we created receiver operating characteristic (ROC) curves. Results: The mtDNA levels in buffy coat were significantly lower than in other sample types. Relative to tumor-adjacent tissue, reduced levels of mtDNA were identified in buffy coat and tumor tissue but not in serum. According to ROC curve analysis, mtDNA levels could be used to distinguish between buffy coat and tumor-adjacent tissue samples with good sensitivity (77%) and specificity (83%). Moreover, mtDNA levels in serum and tumor tissue were positively associated with cancer TMN stage. Conclusions: The mtDNA levels in blood samples may represent a promising, non-invasive biomarker in breast cancer patients. Additional, large-scale validation studies are required to establish the potential use of mtDNA levels in the early diagnosis and monitoring of breast cancer.

Relationship Between Mitochondrial DNA Copy Number, Metabolic Abnormalities and Hepatic Steatosis (지방간 및 대사 인자들과 말초혈액 백혈구의 사립체 DNA copy 수와의 연관성)

  • Kwon, Kil-Young;Jun, Dae-Won
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.6
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    • pp.2093-2098
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    • 2010
  • Insulin resistance plays a central role in fatty liver, a part of the metabolic syndrome. This study examined the relationship between fatty liver, metabolic abnormalities and mitochondrial DNA [mtDNA] copy number in peripheral blood that is correlated with diabetes or metabolic markers. Fatty liver was assessed by questionnaire on alcohol consumption and abdominal ultrasonography. MtDNA copy number in peripheral leukocytes was measured by a real-time quantitative polymerase chain reaction [PCR]. Among 445 subjects, 148 subjects had hepatic steatosis and 297 were controls. mtDNA copy number was significantly lower in fatty liver group in comparison with that of normal finding group. This result is similar in both groups, alcoholic or non-alcoholic fatty liver group. MtDNA copy number was inversely correlated with alanine aminotransferase [ALT], aspartate aminotransferase [AST], gamma-glutamyltransferase [$\gamma$-GTP], body mass index [BMI], waist circumference, diastolic blood pressure, and free fatty acid. MtDNA copy number in peripheral leukocytes was associated with fatty liver and insulin resistance related factors.

Rapid Isolation of Mitochondrial DNA-Depleted Mammalian Cells by Ethidium Bromide and Dideoxycytidine Treatments

  • Yoon, Young Geol;Oh, Yoo Jin;Yoo, Young Hyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.259-265
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    • 2014
  • Mitochondrial DNA (mtDNA)-depleted (${\rho}^0$) cells are often used as mtDNA recipients to study the interaction between the nucleus and mitochondria in mammalian cells. Therefore, it is crucial to obtain mtDNA-depleted cells with many different nuclear backgrounds for the study. Here, we demonstrate a rapid and reliable method to isolate mammalian mtDNA-depleted cells involving treatment with the antimitochondrial agents ethidium bromide (EtBr) and 2',3'-dideoxycytidine (ddC). After a short exposure to EtBr or ddC, followed by rapid clonal isolation, we were able to generate viable mtDNA-depleted cells from mouse and human cells and were able to successfully repopulate them with exogenous mitochondria from platelets isolated from mouse and human blood samples. These mtDNA-depleted cells can be used to characterize the nuclear mitochondrial interactions and to study mtDNA-associated defects in mammalian cells. Our method of isolating mtDNA-depleted cells is practical and applicable to a variety of cell types.

Effect of RFLP Marker of the Mitochondrial DNA D-Loop Region on Milk Production in Korean Cattle (한우 Mitochondrial DNA D-Loop 영역의 RFLP Marker가 산유량에 미치는 영향)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.218-225
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    • 2005
  • This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.