Cell-line misidentification and contamination with microorganisms, such as
mycoplasma, together with instability, both genetic and phenotypic, are among
the problems that continue to affect cell culture. Many of these problems are
avoidable with the necessary foresight, and these Guidelines have been prepared
to provide those new to the field and others engaged in teaching and instruction
with the information necessary to increase their awareness of the problems and
to enable them to deal with them effectively. The Guidelines cover areas such as
development, acquisition, authentication, cryopreservation, transfer of cell
lines between laboratories, microbial contamination, characterisation,
instability and misidentification. Advice is also given on complying with
current legal and ethical requirements when deriving cell lines from human and
animal tissues, the selection and maintenance of equipment and how to deal with
problems that may arise.
Amplification of 8p11-12 is a well-known alteration in human breast cancers but the driving oncogene has not been identified. We have developed a high-resolution comparative genomic hybridization array covering 8p11-12 and analysed 33 primary breast tumors, 20 primary ovarian tumors and 27 breast cancer cell lines. Expression analysis of the genes in the region was carried out by using real-time quantitative PCR and/or oligo-microarray profiling. In all, 24% (8/33) of the breast tumors, 5% (1/20) of the ovary tumors and 15% (4/27) of the cell lines showed 8p11-12 amplification. We identified a 1 Mb segment of common amplification that excludes previously proposed candidate genes. Some of the amplified genes did not show overexpression, whereas for others, overexpression was not specifically attributable to amplification. The genes FLJ14299, C8orf2, BRF2 and RAB11FIP, map within the 8p11-12 minimal amplicon, two have a putative function consistent with an oncogenic role, these four genes showed a strong correlation between amplification and overexpression and are therefore the best candidate driver oncogenes at 8p12.
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