Molecular profiling of cancer.
The objective of molecular profiling of cancer is to determine the differential expression of genes and proteins from human tissue in the progression from normal precursor tissue to preneoplastic tissue to cancer in order to discover diagnostic, prognostic, and therapeutic markers. With the development of high-throughput analytical techniques such as microarrays and 2-D PAGE as well as the development of tools for cell procurement from histological sections such as laser capture microdissection (LCM), it is now possible to perform molecular analyses on specific cell populations from tissue. Since recognition of specific cell populations is critical, there is a need to optimize fixation and embedding not only to improve preservation of biomolecules, but also to maintain excellent histology. We have shown that 70% ethanol fixation of prostate tissue improves the recovery of DNA, RNA, and proteins over routine formalin fixation and maintains histological quality comparable to formalin. There is also a need to develop new technologies in order to expand the range of tissue types that can be analyzed. The development and applications of Layered Expression Scanning (LES) for the molecular analysis of whole tissue sections are discussed.
Gillespie
John W
JW
Science Applications International Corporation, National Cancer Institute, Bethesda, Maryland, USA. jgill@mail.nih.gov
Gannot
Gallya
G
Tangrea
Michael A
MA
Ahram
Mamoun
M
Best
Carolyn J M
CJ
Bichsel
Verena E
VE
Petricoin
Emmanuel F
EF
Emmert-Buck
Michael R
MR
Chuaqui
Rodrigo F
RF
D004273:DNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D006801:Humans; D007834:Lasers; D008297:Male; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism; D009369:Neoplasms / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D011471:Prostatic Neoplasms / Q000737:chemistry* / Q000378:metabolism* / Q000473:pathology; D040901:Proteomics; D012334:RNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D013048:Specimen Handling
D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm