The Effect of Elapsed Time on the Quantity of mRNA in Skin: A Study to Evaluate the Potential Forensic Use of mRNA to Determine the Postmortem Interval

다국어 초록 (Multilingual Abstract)

Ribonucleic acid (RNA) has potential use in forensic science for the determination of postmortem interval. We report the first study on serial sampling of messenger RNA (mRNA) from surgical specimens to determine if there is a correlation between mRNA quantity and elapsed time. Skin tissues were collected from modified radical mastectomy specimens. After a defined period of time, bisected skin sections were cut and frozen in liquid nitrogen. Serial collection of the specimens was conducted, and frozen sections were obtained from all samples. Quantitative real-time reverse transcriptionpolymerase chain reaction was performed using the extracted RNA to measure the transcriptional activity of 2 selected housekeeping genes. The selected loci were mRNA sequences that exhibited time-dependent quantitative changes in a previous study. We collected 44 samples from 9 different patients, with 3-10 samples collected per patient. The amount of mRNA transcripts present in the serial samples showed a weak time-dependent correlation trend only in some cases. Further studies to evaluate different target mRNA sequences are necessary, as is exploration of additional methods to evaluate mRNA transcript degradation.

참고문헌 (Reference)

1 Harrison PJ, "The relative importance of premortem acidosis and postmortem interval for human brain gene expression studies: selective mRNA vulnerability and comparison with their encoded proteins" 200 : 151-154, 1995

2 Hardy JA, "The patients dying after long terminal phase have acidotic brains; implications for biochemical measurements on autopsy tissue" 61 : 253-264, 1985

3 Barrachina M, "TaqMan PCR assay in the control of RNA normalization in human post-mortem brain tissue" 49 : 276-284, 2006

4 Li JZ, "Systematic changes in gene expression in postmortem human brains associated with tissue pH and terminal medical conditions" 13 : 609-616, 2004

5 Heinrich M, "Successful RNA extraction from various human postmortem tissues" 121 : 136-142, 2007

6 Trotter SA, "Stability of gene expression in postmortem brain revealed by cDNA gene array analysis" 942 : 120-123, 2002

7 Heinrich M, "Realtime PCR detection of five different “endogenous control gene” transcripts in forensic autopsy material" 1 : 163-169, 2007

8 Zhao D, "Real-time RT-PCR quantitative assays and postmortem degradation proles of erythropoietin, vascular endothelial growth factor and hypoxia-inducible factor 1 alpha mRNA transcripts in forensic autopsy materials" 8 : 132-136, 2006

9 Bauer M., "RNA in forensic science" 1 : 69-74, 2007

10 Preece P, "Quantifying mRNA in postmortem human brain: influence of gender, age at death, postmortem interval, brain pH, agonal state and inter-lobe mRNA variance" 118 (118): 60-71, 2003

11 Bauer M, "Quantification of mRNA degradation as possible indicator of postmortem interval-- a pilot study" 5 : 220-227, 2003

12 Bauer M, "Quantification of RNA degradation by semi-quantitative duplex and competitive RTPCR: a possible indicator of the age of bloodstains?" 138 : 94-103, 2003

13 Madea B., "Is there recent progress in the estimation of the postmortem interval by means of thanatochemistry?" 151 : 139-149, 2005

14 Di Nunno NR, "Is flow cytometric evaluation of DNA degradation a reliable method to investigate the early postmortem period?" 19 : 50-53, 1998

15 Castensson A, "High-resolution quantification of specific mRNA levels in human brain autopsies and biopsies" 10 : 1219-1229, 2000

16 Yasojima K, "High stability of mRNAs postmortem and protocols for their assessment by RT-PCR" 8 : 212-218, 2001

17 Boy SC, "Flow cytometric evaluation of postmortem pulp DNA degradation" 24 : 123-127, 2003

18 Cina SJ, "Flow cytometric evaluation of DNA degradation: a predictor of postmortem interval?" 15 : 300-302, 1994

19 Udvardi MK, "Eleven golden rules of quantitative RT-PCR" 20 : 1736-1737, 2008

20 Inoue H, "Degradation profile of mRNA in a dead rat body: basic semi-quantification study" 130 : 127-132, 2002

21 Sharova LV, "Database for mRNA half-life of 19977 genes obtained by DNA microarray analysis of pluripotent and differentiating mouse embryonic stem cells" 16 : 45-58, 2009

22 Swift GH, "Assessment of RNA quality by semi-quantitative RT-PCR of multiple regions of a long ubiquitous mRNA" 28 : 524-526, 2000

23 Partemi S, "Analysis of mRNA from human heart tissue and putative applications in forensic molecular pathology" 203 : 99-105, 2010

24 Ishida K, "A quantitative RT-PCR assay of surfactant-associated protein A1 and A2 mRNA transcripts as a diagnostic tool for acute asphyxial death" 4 : 7-12, 2002

25 Anderson S, "A method for determining the age of a bloodstain" 148 : 37-45, 2005