A Simple Bioluminescence Imaging Method for Studying Cancer Cell Growth and Metastasis after Subcutaneous Injection of Lewis Lung Carcinoma Cells in Syngeneic C57BL/6 Mice

Abstract

In vivo imaging of cancer cell growth and invasion is instrumental in studying cancer cell behavior and in developing effective anticancer agents. In this ROS Protocols article, we report the experimental protocol and steps involving the implantation of luciferase-expressing Lewis lung carcinoma (LLC) cells in normal syngeneic C57BL/6 mice. Using the Berthold NightOwl LB981 in vivo imaging system, we observe the time-dependent growth and invasion of the lung cancer cells following subcutaneous injection of luciferase-expressing LLC cells. The three-dimensional image and counts of photon emission of the tumor mass are obtained to estimate the relative size of the tumor. Ex vivo imaging of the isolated lungs supplemented with D-luciferin and adenosine triphosphate (ATP) is obtained to determine lung metastasis of the LLC cells. The LLC cell load in entire mouse lungs is further determined by quantitative bioluminometry with a concurrently run standard curve of the number of LLC cells versus bioluminescence intensity. This in vivo imaging system in live mice, in combination with ex vivo imaging of isolated lungs as well as quantitative bioluminometry of target tissues, may provide important information on the in vivo cancer cell dynamics in immunocompetent syngeneic C57BL/6 mice and offer a valuable tool for studying experimental anticancer agents, including redox-modulating compounds, which are promising anticancer modalities.

High COX-2 expression contributes to a poor prognosis through the inhibition of chemotherapy-induced senescence in nasopharyngeal carcinoma

“…4D that the numbers of SA-β-gal-positive cells were increased in fibroblasts from COX-2 −/− mice (C57BL/6 background), which suggested that senescence was induced in the COX-2 −/− mouse model, this prompted us to investigate the role of senescence in the mice with the same genetic C57BL/6 background following treatment with NS-398. As the LLC cell lines can lead to tumor bearing C57BL/6 WT mice ( 31 ), while none of the NPC cell lines could, we selected the LLC line for use in this in vivo experiment. The COX-2 inhibitor, NS-398, was employed in the mouse model established by subcutaneous inoculation with 1×10 6 LLC cells into C57BL/6 mouse ( Fig.…”

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention

“…This was further analyzed by NightOWL LB 983 in vivo imaging system, where the uorescent detection potential of FusionRed transfected DT0840 and DT0846 cell lines were veri ed. Small cell numbers were su cient to monitor the FusionRed labeled cells in the whole body imager [59]. For in vivo imaging studies a cell number from 1 × 10 6 cells per tumor is detectable with this setting already in early tumor stages [60].…”

Electrostatic reaction for the detection of circulating tumor cells as a potential diagnostic biomarker for metastasis in solid tumor

“…Therefore, various mouse models have been developed in CTCs capture studies, including syngeneic models (4T1 murine breast cancer cell line) [25], transgenic models (overexpressing EGFR) [26], and patient-derived xenografts (PDX, triple negative breast cancer patient-derived xenograft) [27], etc. In our study, we established a syngeneic model using murine cancer cells LLC1 and B16F10, which was chosen due to the ability to quickly metastasize to the lungs [15,28]. In vivo metastasis models are increasingly being used to study the correlation between the count of CTCs and tumor progression.…”