Surgical injury enhances the expression of genes that mediate breast cancer metastasis to the lung.

OBJECTIVE: This study was conducted to test the hypothesis that surgery induces changes at the expression level of genes implicated in metastasis, thus leading to accelerated postoperative metastatic tumor growth. SUMMARY BACKGROUND DATA: Surgical resection of the primary tumor is a necessary and effective treatment for breast cancer patients. However, studies from both animals and humans have shown that surgery potentiates the growth of minimal residual neoplastic disease. METHODS: : Female BALB/c mice were inoculated with metastatic murine mammary adenocarcinoma 4T1-green fluorescent protein (GFP) cells in the mammary fat pad (3 × 105/mouse), and divided into a surgery group (n = 12) in which the flank tumor was completely resected after 21 day growth and a control (no surgery) group (n = 12). Metastatic tumor burden was assessed by both macroscopic metastatic nodule count and clonogenic assay. Mitotic and apoptotic indices were established using a combination of hematoxylin-eosin histology and Ki-67 immunohistochemistry. Green fluorescent protein (GFP) expressing tumor cells were isolated using FACS sorting, and RNA was extracted. The RT² Profiler PCR Array mouse Cancer Pathway Finder was used to determine and compare the mRNA levels of 84 genes involved in metastasis in both groups. RESULTS: Excision of the primary tumor was associated with increased systemic metastatic burden (P = 0.001). Postoperative metastases exhibited increased proliferation (P = 0.001), but no reduction in apoptosis. The quantitative real-time polymerase chain reaction array data indicate that surgery significantly upregulated the expression of Itgb3, Egfr, Hgf, Igf1, Pdgfb, Tnfα, Vegfa, Vegfc, and MMP9 genes, and led to the down regulation of Cdkn2a, Cdh1, and Syk genes. Increased expression of ITGB3 and MMP9 was further confirmed at the protein level by Western blot. CONCLUSIONS: Removal of the primary tumor led to a progressive phenotype of lung metastases that exhibited upregulation of genes involved in adhesion, invasion, and angiogenesis.

The targeting of phosphoinositide-3 kinase attenuates pulmonary metastatic tumor growth following laparotomy.

OBJECTIVE: We aimed to characterize a potential role for phosphatidylinositol 3-kinase (PI3k) in leading to accelerated postoperative metastatic tumor growth. BACKGROUND: PI3k enhances tumor cell survival in part by phosphorylating Akt and reducing apoptosis. Postoperatively, apoptosis is reduced within local recurrences and distant metastases. This reduction is associated with the phenomenon of accelerated postoperative tumor growth. METHODS: Balb/c mice underwent a tail vein injection of 1x10 metastatic murine mammary adenocarcinoma 4T1 cells. Animals were divided into the following treatment groups (n=10/group): group A, controls; group B, DMSO intraperitoneally (IP) daily from days 14 to 21; group C, IP LY294002 daily from days 14 to 21; group D, laparotomy only; group E, laparotomy followed by IP DMSO for 7 days; and group F, laparotomy followed by LY294002 IP for 7 days. All laparotomies were performed on day 14. Animals were killed at day 28. Metastatic tumor burden was assessed using the lung/body weight ratio and a histologic metastatic index. Mitotic counts and apoptotic indices were established using a combination of hematoxylin and eosin histology and TUNEL immunohistochemistry. A parallel survival study was performed, and PI3k activity was assessed using western blots for phospho-Akt. RESULTS: Laparotomy was associated with increased systemic tumor burden (P=0.001). Postoperatively, LY294002 significantly attenuated metastatic tumor growth (P<0.001). Effective PI3k inhibition was confirmed by demonstrating a reduced Akt phosphorylation. Moreover, PI3k inhibition led to reduced proliferation, increased apoptosis (P<0.001), and enhanced postoperative survival (P<0.001). CONCLUSIONS: Targeting PI3k with postoperative LY294002 significantly attenuates the acceleration in postoperative metastatic tumor growth seen following laparotomy.

Phosphoinositide 3-kinase accelerates postoperative tumor growth by inhibiting apoptosis and enhancing resistance to chemotherapy-induced apoptosis. Novel role for an old enemy.

Tumor removal remains the principal treatment modality in the management of solid tumors. The process of tumor removal may potentiate the resurgent growth of residual neoplastic tissue. Herein, we describe a novel murine model in which flank tumor cytoreduction is followed by accelerated local tumor recurrence. This model held for primary and recurrent tumors generated using a panel of human and murine (LS174T, DU145, SW480, SW640, and 3LL) cell lines and replicated accelerated tumor growth following excisional surgery. In investigating this further, epithelial cells were purified from LS174T primary and corresponding recurrent tumors for comparison. Baseline as well as tumor necrosis factor apoptosis-inducing ligand (TRAIL)-induced apoptosis were significantly reduced in recurrent tumor epithelia. Primary and recurrent tumor gene expression profiles were then compared. This identified an increase and reduction in the expression of p110gamma and p85alpha class Ia phosphoinositide 3-kinase (PI3K) subunits in recurrent tumor epithelia. These changes were further confirmed at the protein level. The targeting of PI3K ex vivo, using LY294002, restored sensitivity to TRAIL in recurrent tumor epithelia. In vivo, adjuvant LY294002 prolonged survival and significantly attenuated recurrent tumor growth by greatly enhancing apoptosis levels. Hence, PI3K plays a role in generating the antiapoptotic and chemoresistant phenotype associated with accelerated local tumor recurrence.