Abstract 5091: Proteomic study using a newly established metastatic lung cancer model

Abstract

Abstract To understand metastasis of lung cancer, we established a highly metastatic pulmonary large cell carcinoma cell line (801BL) through 2 rounds of in vivo selection using a nude mouse xenograft model. In the first round, low-metastatic human large cell lung cancer cells (801D cell line) were subcutaneously injected into the flank of the mice. Metastatic tumor cells were collected from mouse brain and re-injected into the mice flank in the second round. A highly metastatic cell line 801BL was established from the metastatic cells in the lung. A PCR-based satellite tandem repeat (STR) analysis confirmed the same genomic background of the newly established metastatic cell line 801BL as the non-metastatic counterpart 801C and low-metastatic counterpart 801D cell lines. A xenografted mouse model showed 100% of the mice (8 out of 8 mice) injected subcutaneously with 801BL developed lung metastatic tumors, while none of the mice injected with 801C cells developed lung metastasis (p<0.0001). The metastatic cells present a different cancer stem cell-like morphological feature compared with the 801C and 801D cell lines. Their invasive capability is also increased by 3.47±0.44 fold compared to the non-metastatic counterpart (801C) in a matrigel invasion assay. A stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomic study was conducted by comparing the isotopic labeled 801BL cells with non-labeled 801C cells. Bioinformatic analysis using Ingenuity software (http://ingenuity.com) revealed significant alterations in several dominant cell signal networks including cell morphology (p-value:2.80E-09 - 6.83E-03), cellular function and maintenance (p-value:1.08E-08 - 6.83E-03), DNA replication, recombination, and repair (p-value:1.08E-08 - 6.83E-03), and cell death (p-value: 1.88E-08 - 6.83E-03), between the highly metastatic cell line (801BL) and its non-metastatic counterpart (801C). Western blot assay of several proteins from each signaling network confirmed the proteomic findings. Since the highly metastatic cell line and its non-metastatic counterpart share the identical genetic background, the model based on this cell line provides a powerful tool for the study of the mechanisms underlying lung cancer metastasis. (This work is supported by grant from DOD W81XWH-07-1-0306). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5091. doi:1538-7445.AM2012-5091