Measuring growth and gene expression dynamics of tumor-targeted S. typhimurium bacteria.

Publication Type:

Journal Article

Source:

J Vis Exp, Issue 77, p.e50540 (2013)

Keywords:

Animals, Cell Line, Tumor, Female, Gene Expression Profiling, Heterografts, Humans, Mice, Mice, Nude, Microbiological Techniques, Ovarian Neoplasms, Salmonella typhimurium

Abstract:

The goal of these experiments is to generate quantitative time-course data on the growth and gene expression dynamics of attenuated S. typhimurium bacterial colonies growing inside tumors. We generated model xenograft tumors in mice by subcutaneous injection of a human ovarian cancer cell line, OVCAR-8 (NCI DCTD Tumor Repository, Frederick, MD). We transformed attenuated strains of S. typhimurium bacteria (ELH430:SL1344 phoPQ- (1)) with a constitutively expressed luciferase (luxCDABE) plasmid for visualization(2). These strains specifically colonize tumors while remaining essentially non-virulent to the mouse(1). Once measurable tumors were established, bacteria were injected intravenously via the tail vein with varying dosage. Tumor-localized, bacterial gene expression was monitored in real time over the course of 60 hours using an in vivo imaging system (IVIS). At each time point, tumors were excised, homogenized, and plated to quantitate bacterial colonies for correlation with gene expression data. Together, this data yields a quantitative measure of the in vivo growth and gene expression dynamics of bacteria growing inside tumors.

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