TRIzol and Alu qPCR-based quantification of metastatic seeding within the skeleton.
Current methods for detecting disseminated tumor cells in the skeleton are limited by expense and technical complexity. We describe a simple and inexpensive method to quantify, with single cell sensitivity, human metastatic cancer in the mouse skeleton, concurrently with host gene expression, using TRIzol-based DNA/RNA extraction and Alu sequence qPCR amplification. This approach enables precise quantification of tumor cells and corresponding host gene expression during metastatic colonization in xenograft models.
1] Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Preston Campbell
1] Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Mulcrone
Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Masood
1] Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Karolak
1] Department of Veterans Affairs (VISN 9), Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Merkel
1] Department of Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Hebron
1] Department of Pathology, Microbiology, and Immunology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Zijlstra
1] Department of Veterans Affairs (VISN 9), Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Sterling
1] Department of Pharmacology, Vanderbilt University, Nashville, Tennessee, United States of America [2] Vanderbilt Center for Bone Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
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Elefteriou
D020087:Alu Elements / Q000235:genetics; D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics*; D001859:Bone Neoplasms / Q000235:genetics* / Q000473:pathology / Q000556:secondary*; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D020869:Gene Expression Profiling / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D006146:Guanidines / Q000737:chemistry; D051379:Mice; D008819:Mice, Nude; D010636:Phenols / Q000737:chemistry; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D005819:Genetic Markers; D006146:Guanidines; D010636:Phenols; D012334:RNA, Neoplasm; C411644:trizol