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Postmortem interval determination using mRNA markers and DNA normalization

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Abstract

Postmortem interval (PMI) determination is an important part of criminal investigations, but it is still subject to uncertainty. Degradation of mRNA in PMI determination has been studied in decays; however, some studies have reported no correlation between PMI and RNA degradation. Thus, we aimed to determine whether RNA quantity was correlated with PMI. Heart and brain tissues were separated from a mouse model of a 0–48 h PMI with 29 time points. We then coextracted the DNA and RNA in one tube with Bioteke coextraction kits and selected some mRNA markers associated with cell oxygen deprivation and apoptosis as target genes, such as hypoxia-associated factor (HAF), apoptosis-inducing factor (AIF), hypoxia-inducible factor 2 alpha (HIF2a), and factor inhibiting HIF (FIH). We measured the quantity of these markers using real-time quantitative PCR (qPCR), and Caspase-3 DNA and 18S were each used for normalization. The results showed that in the heart tissue, the degradation of HIF2a, AIF, and FIH was correlated with PMI, as was the degradation of HIF2a, FIH, and AIF in brain tissue when normalized with Caspase-3 DNA. However, when normalized with 18S, only the degradation of HIF2a in brain tissue was correlated with PMI. Interestingly, the quantity of HAF in brain tissue was found to increase after death with either 18S or Caspase-3 DNA normalization, and it was significantly correlated with 0–48 h PMI. These results indicated that mRNA quantity can be used to determine PMI and that Caspase-3 DNA is feasible for PMI estimation. In summary, we established mathematical models for PMI determination using multiple mRNA markers and multiple tissues and further studies are needed to validate and investigate these markers and mathematical models in human tissues.

Duo Peng and Meili Lv contributed equally to this work.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 81471827, No. 81671871), the Science Foundation for the Excellent Youth Scholars of Sichuan University (No. 2082604174158), and the Science Foundation of Sichuan Public Security Department (2017SCLL04).

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Author notes

  1. Duo Peng and Meili Lv contributed equally to this work.

Authors and Affiliations

  1. Department of Forensic Genetics, West China School of Basic Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China

    Duo Peng, Meili Lv, Zhilong Li, Huan Tian, ShengQiu Qu, Weibo Liang & Lin Zhang

  2. Department of Forensic Medicine, North Sichuan Medical College, Nanchong, 637000, Sichuan, China

    Bo Jin

  3. Sichuan Police College, Luzhou, 646000, Sichuan, China

    Bing Long

Authors
  1. Duo Peng
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  2. Meili Lv
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  3. Zhilong Li
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  4. Huan Tian
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  5. ShengQiu Qu
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Corresponding authors

Correspondence to Weibo Liang or Lin Zhang.

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The experimental procedures and the animal use and care protocols were approved by the Ethics Committee of Sichuan University and Experimental Animal Care and Use Committee of Sichuan University, China.

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The authors declare that they have no conflict of interest.

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Cite this article

Peng, D., Lv, M., Li, Z. et al. Postmortem interval determination using mRNA markers and DNA normalization. Int J Legal Med 134, 149–157 (2020). https://doi.org/10.1007/s00414-019-02199-7

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  • DOI: https://doi.org/10.1007/s00414-019-02199-7

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