Detection of tumor messenger RNA in the serum of patients with malignant melanoma.
Serum RNases are known to be elevated in patients with cancer. Consequently, it is not clear whether human mRNA with sufficient integrity as to permit reverse transcription-PCR (RT-PCR) amplification is detectable in serum. We examined serum from six patients with malignant melanoma for human tyrosinase mRNA using RT-PCR. Serum from 20 normal volunteers served as controls. Tyrosinase mRNA could be demonstrated in serum from four of the six melanoma patients with detection by gel electrophoresis and confirmation by blotting amplified product to a tyrosinase-specific probe. The serum remained tyrosinase mRNA positive, even if passed through a 0.45 microm filter prior to RNA extraction, indicating that the mRNA was extracellular at the time of extraction. Tyrosinase mRNA could not be detected in any control serum (0 of 20 individuals). The presence and integrity of amplifiable RNA was confirmed in all serum specimens (patients and controls) by RT-PCR amplification of c-abl mRNA. Amplifiable RNA could be demonstrated regardless of whether serum was freshly drawn or stored frozen for several years. We conclude that human mRNA can be extracted and amplified from serum. The ability to amplify tumor mRNA from serum may have important utility in cancer diagnostics and monitoring.
Kopreski
M S
MS
OncoMEDx, Inc., Columbia, Maryland, USA.
Benko
F A
FA
Kwak
L W
LW
Gocke
C D
CD
D015139:Blotting, Southern; D004587:Electrophoresis, Agar Gel; D005615:Freezing; D006801:Humans; D008545:Melanoma / Q000097:blood / Q000201:enzymology*; D014442:Monophenol Monooxygenase / Q000097:blood / Q000235:genetics*; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000097:blood* / Q000737:chemistry; D012334:RNA, Neoplasm / Q000097:blood* / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014442:Monophenol Monooxygenase