| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Received on November 4, 2005
Accepted on March 16, 2006
Molecular Diagnostics and Genetics |
1 Dental Research Institute, UCLA, Los Angeles, CA
2 Dental Research Institute and Department of Urology, David Geffen School of Medicine at UCLA, Los Angeles, CA
3 Dental Research Institute, UCLA School of Dentistry, Johnson Comprehensive Cancer Center, Division of Head and Neck Surgery/Otolaryngology, and Henry Samueli School of Engineering, UCLA, Los Angeles, CA
* To whom correspondence should be addressed. E-mail: dtww{at}ucla.edu.
Background: We have previously shown that human mRNAs are present in saliva and can be used as biomarkers of oral cancer. In this study, we analyzed the integrity, sources, and stability of salivary RNA.
Methods: We measured the integrity of salivary RNA with reverse transcription followed by PCR (RT-PCR) or RT-quantitative PCR (RT-qPCR). To study RNA entry sites into the oral cavity, we used RT-PCR analysis of salivary RNA from the 3 major salivary glands, gingival crevice fluid, and desquamated oral epithelial cells. We measured stability of the salivary 
-actin mRNA by RT-qPCR of salivary RNA incubated at room temperature for different periods of time. We measured RNA association with other macromolecules by filtering saliva through pores of different sizes before performing RT-qPCR. To assess RNA-macromolecule interaction, we incubated saliva with Triton X-100 for different periods of time before performing RT-qPCR.
Results: In most cases, we detected partial- to full-length salivary mRNAs and smaller amounts of middle and 3' gene amplicons compared with the 5'. RNA was present in all oral fluids examined. Endogenous salivary -actin mRNA degraded more slowly than exogenous -actin mRNA, with half-lives of 12.2 and 0.4 min, respectively (P <0.001). Salivary RNA could not pass through 0.22 or 0.45 µm pores. Incubation of saliva with Triton X-100 accelerated degradation of salivary RNA.
Conclusions: Saliva harbors both full-length and partially degraded forms of mRNA. RNA enters the oral cavity from different sources, and association with macromolecules may protect salivary RNA from degradation.
The following articles in journals at HighWire Press have cited this article:
|
|
 |
|
  S. Huang, H. Li, X. Ding, and C. Xiong Presence and Characterization of Cell-Free Seminal RNA in Healthy Individuals: Implications for Noninvasive Disease Diagnosis and Gene Expression Studies of the Male Reproductive System Clin. Chem., November 1, 2009; 55(11): 1967 - 1976. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. J. Park, H. Zhou, D. Elashoff, B. S. Henson, D. A. Kastratovic, E. Abemayor, and D. T. Wong Salivary microRNA: Discovery, Characterization, and Clinical Utility for Oral Cancer Detection Clin. Cancer Res., September 1, 2009; 15(17): 5473 - 5477. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Wei, P. Patel, W. Liao, K. Chaudhry, L. Zhang, M. Arellano-Garcia, S. Hu, D. Elashoff, H. Zhou, S. Shukla, et al. Electrochemical Sensor for Multiplex Biomarkers Detection Clin. Cancer Res., July 1, 2009; 15(13): 4446 - 4452. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Palanisamy, N.J. Park, J. Wang, and D.T. Wong AUF1 and HuR Proteins Stabilize Interleukin-8 mRNA in Human Saliva Journal of Dental Research, August 1, 2008; 87(8): 772 - 776. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Koka, T. J. Beebe, S. P. Merry, R. S. DeJesus, L. D. Berlanga, A. L. Weaver, V. M. Montori, and D. T. Wong The Preferences of Adult Outpatients in Medical or Dental Care Settings for Giving Saliva, Urine or Blood for Clinical Testing J Am Dent Assoc, June 1, 2008; 139(6): 735 - 740. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Hu, B. G. Zimmermann, H. Zhou, J. Wang, B. S. Henson, W. Yu, D. Elashoff, G. Krupp, and D. T. Wong Exon-Level Expression Profiling: A Comprehensive Transcriptome Analysis of Oral Fluids Clin. Chem., May 1, 2008; 54(5): 824 - 832. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. T. Wong Salivary Transcriptome Clin. Cancer Res., February 15, 2007; 13(4): 1350 - 1350. [Full Text] [PDF]
|
|
|
|
 |
|
 L. Seugnet, J. Boero, L. Gottschalk, S. P. Duntley, and P. J. Shaw Identification of a biomarker for sleep drive in flies and humans PNAS, December 26, 2006; 103(52): 19913 - 19918. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
