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Mutational Analysis of the Mitochondrial DNA Displacement-Loop Region in Human Retinoblastoma with Patient Outcome

  • Original Article
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Pathology & Oncology Research

Abstract

Alteration in mitochondrial DNA plays an important role in the development and progression of cancer. The Displacement Loop (D-loop) region of mitochondrial DNA (mtDNA) is the regulatory region for its replication and transcription. Therefore, we aimed to characterize mutations in the D-loop region of mitochondrial DNA along with the morphological changes and analyzed their impact on survival in retinoblastoma patients. mtDNA D-loop region was amplified by Nested-Polymerase Chain Reaction (Nested-PCR) and mutations were analyzed in 60 tumor samples from retinoblastoma patients by DNA sequencing. Transmission electron microscopy was performed on 5 retinoblastoma specimens. Mutations were correlated with clinical, histopathological parameters and patient survival. D-loop mutations were found in total of 52/60 (86.6%) patients. The most common mutations were T to C and C to T followed by A to G. There were 5.81% mutations which were not previously reported in the MITOMAP database. A73G (83.33%) were the most frequent mutations found in our cases and it was statistically significant with poor tumor differentiation and age. In addition, this study was further analyzed for morphological changes in retinoblastoma that had disorganized, swollen and less numbers of mitochondria on electron microscopy. This is the first study showing high frequency of mtDNA mutation which might be due to abnormal morphology of mitochondria in retinoblastoma. Our results indicate that pathogenic mtDNA D-loop mutations may be involved in tumorigenesis of retinoblastoma tumor.

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Fig. 1
Fig. 2
Fig. 3: Kaplan Meier Curve showing Overall Survival Rate of Retinoblastoma Patients based on mutations at Frequent Sites.
Fig. 4: Ultrastructural Morphology of Mitochondria in Retinoblastoma Tumor Samples.

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Funding

This study was funded by Indian Council of Medical Research, New Delhi, INDIA (Award no: 5/4/6/09/Oph/2012-NCD II).

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Author notes

  1. Lata Singh and Seema Kashyap contributed equally to this work.

Authors and Affiliations

  1. Department of Ocular Pathology, Dr. R. P. Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India

    Lata Singh, Seema Sen & Seema Kashyap

  2. Functional Genomics Unit, Institute of Genomics and Integrative Biology, Mall Road, New Delhi, India

    Neeru Saini

  3. Department of Ophthalmology, Dr. R. P. Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India

    Neelam Pushker

  4. Department of Medical Oncology, IRCH, All India Institute of Medical Sciences, New Delhi, India

    Sameer Bakhshi

  5. Department of Anatomy, All India Institute of Medical Sciences, New Delhi, India

    Tapas C. Nag

Authors
  1. Lata Singh
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  2. Neeru Saini
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  3. Neelam Pushker
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  4. Sameer Bakhshi
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  5. Seema Sen
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  6. Tapas C. Nag
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  7. Seema Kashyap
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Contributions

LS was responsible for designing and executing the experiment. NS helps in sequencing analysis. SK and SS did the histopathological evaluation. TCN did the electron microscopy evaluation. SB was responsible for providing follow up. NP was responsible proving samples.

Corresponding authors

Correspondence to Lata Singh or Seema Kashyap.

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No potential conflicts of interest with any author.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Cite this article

Singh, L., Saini, N., Pushker, N. et al. Mutational Analysis of the Mitochondrial DNA Displacement-Loop Region in Human Retinoblastoma with Patient Outcome. Pathol. Oncol. Res. 25, 503–512 (2019). https://doi.org/10.1007/s12253-018-0391-y

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  • DOI: https://doi.org/10.1007/s12253-018-0391-y

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