Hypertonic Saline Attenuates the Pro-metastatic Effects of LPS by Reducing Tumor Cell Migration, Proliferation and MMP-9 Expression

BACKGROUND: Lipopolysaccharide (LPS) promotes tumor metastases. The aim of this study was to determine the ability of a hypertonic environment to attenuate the pro metastatic properties of LPS both in vitro, and in vivo. METHODS: LPS stimulated, and unstimulated, 4T1 tumor cells were cultured in either an isotonic or hypertonic environment. The effect on invasion, migration, pro-matellomatrixproteinase 9 (proMMP-9) expression, proliferation, and microscopic cell structure was assessed. Lung metastases were induced in C57 mice with systemic hypertonicity in unstimulated and stimulated mice. The metastatic burden was assessed by estimation of lung/body weight ratio, pleural nodules and clonogenic assay. RESULTS: In vitro, a hypertonic environment reduced proMMP-9 expression (0.012 versus 1.16, P < 0.001) invasion (0.06 versus 0.119, P = 0.005), tumor cell proliferation (0.035 versus 0.041, P = 0.001), while inducing structural changes to tumor cells reducing overall cell volume. In vivo, the induction of transient systemic hypertonicity reduced metastatic burden as demonstrated by reduced lung nodules (4 versus 8, P = 0.004) and colonies on clonogenic assay (12 versus 43, P = 0.04). CONCLUSION: The in vitro exposure of tumor cells to a hypertonic environment reduces tumor cell migration and proliferation. Transient systemic hypertonicity can reduce the metastatic burden following intra-operative exposure to LPS in vivo.