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Human-specific GAPDH RT-qPCR is an accurate and sensitive method of xenograft metastasis quantification

Metastasis is the primary cause of cancer-related mortality. Experimental models that accurately reflect changes in metastatic burden are essential tools for developing treatments and to gain a better understanding of disease. Murine xenograft tumor models mimic the human scenario and provide a platform for metastasis analyses. An ex vivo quantitative method, gaining favor for its ease and accuracy is reverse transcription-qualitative polymerase chain reaction (RT-qPCR); however, it is currently unclear how well this method correlates with gold-standard histological analysis and its use has required detection of overexpressed exogenous genes. We have introduced a variation of the RT-qPCR method; human-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDH) RT-qPCR, which allows quantification of metastasis in xenograft models, without the requirement of overexpressed exogenous genes. This makes the method easily amenable to many xenograft models without alteration of the cancer cells. We determined that the method is able to detect a few human cells within abundant mouse lung tissue. Further, the human-specific GAPDH RT-qPCR is more sensitive and correlates with histological analysis in terms of determining relative metastatic burden, suggesting that human-specific GAPDH RT-qPCR could be used as a primary method for quantification of disseminated human cells in murine xenograft models.Graphical AbstractDownload : Download high-res image (111KB)Download : Download full-size image

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