Fluorescence in situ hybridization (FISH): an increasingly demanded tool for biomarker research and personalized medicine

Abstract

Extensive studies of the genetic aberrations related to human diseases conducted over the last two decades have identified recurrent genomic abnormalities as potential driving factors underlying a variety of cancers. Over the time, a series of cutting-edge high-throughput genetic tests, such as microarrays and next-generation sequencing, have been developed and incorporated into routine clinical practice. Although it is a classical low-throughput cytogenetic test, fluorescence in situ hybridization (FISH) does not show signs of fading; on the contrary, it plays an increasingly important role in detecting specific biomarkers in solid and hematologic neoplasms and has therefore become an indispensable part of the rapidly developing field of personalized medicine. In this article, we have summarized the recent advances in FISH application for both de novo discovery and routine detection of chromosomal rearrangements, amplifications, and deletions that are associated with the pathogenesis of various hematopoietic and non-hematopoietic malignancies. In addition, we have reviewed the recent developments in FISH methodology as well.

Figure 1

The three subclones from the ascites cytospin sample of a progressive epithelial ovarian cancer patient. BAC probes containing the c-myc, Rb1, Chk2, p53 or BRCA1 genes were labeled with Spectrum Green, PF555 (red), PF590 (orange), HyPer5 (purple) or PF415 (blue), respectively. The mixed probes were hybridized with the ascites cytospin sample from a progressive epithelial ovarian cancer patient. The results revealed that there were three subclones showing distinct combinations of signal patterns for the five selected genes. The details of the molecular profiling are shown in Table 1.

Figure 2

The three subclones obtained from a cancer stem-cell preparation for a patient with low-differentiated ovarian adenocarcinoma. BAC probes containing c-myc, Rb1, Chk2, p53 or BRCA1 were labeled with Spectrum Green, PF555 (red), PF590 (orange), HyPer5 (purple) or PF415 (blue), respectively. The mixed probes were hybridized with the cancer stem-cell preparation sample from a low-differentiated ovarian adenocarcinoma patient. The results revealed that there were three subclones showing distinct combinations of signal patterns for the five selected genes. The details of the molecular profiling are shown in Table 2.

Figure 3

The six subclones from the bone marrow sample taken from an ALL patient. BAC probes containing the TEL, AML1, PAX5, p16 or IKZF1 genes were labeled with SpectrumGreen, PF555 (red), PF590 (orange), HyPer5 (purple) or PF415 (blue), respectively. The mixed probes were hybridized with the bone marrow sample of an ALL patient. The results revealed that there were six subclones showing distinct combinations of the signal patterns for the five selected genes. The details of the molecular profiling are shown in Table 3.

Figure 4

The clonal components of bone marrow samples taken from an ALL patient upon the initial diagnosis and after chemotherapy. BAC probes containing the TEL, AML1, PAX5, p16 or IKZF1 genes were labeled with Spectrum Green, PF555 (red), PF590 (orange), HyPer5 (purple) or PF415 (blue), respectively. The mixed probes are hybridized with the bone marrow samples upon the initial diagnosis and after thermotherapy of an ALL patient. The results revealed that there are distinct subclones upon the initial diagnosis and after chemotherapy, which showed different combinations of signal patterns for the five selected genes. The details of the molecular profiling are shown in Table 4.