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Transforming growth factor‐β ( TGF ‐β) is a potent growth inhibitor in normal epithelial cells. However, a number of malignant tumors produce excessive amounts of  TGF ‐β, which affects the tumor‐associated microenvironment by furthering the progression of tumorigenicity. Although it is known that the tumor‐associated microenvironment often becomes hypoxic, how hypoxia influences  TGF ‐β signaling in this microenvironment is unknown. We investigated whether  TGF ‐β signaling is influenced by long‐term exposure to hypoxia in Lewis lung carcinoma ( LLC ) cells. When the cells were exposed to hypoxia for more than 10 days, their morphology was remarkably changed to a spindle shape, and  TGF ‐β‐induced Smad2 phosphorylation was enhanced. Concomitantly,  TGF ‐β‐induced transcriptional activity was augmented under hypoxia, although  TGF ‐β did not influence the activity of a hypoxia‐responsive reporter. Consistently, hypoxia influenced the expression of several  TGF ‐β target genes. Interestingly, the expressions of  TGF ‐β type I receptor (Tβ RI ), also termed activin receptor like kinase‐5 ( ALK 5), and  TGF ‐β1 were increased under the hypoxic condition. When we monitored the hypoxia‐inducible factor‐1 ( HIF ‐1) transcriptional activity by use of green fluorescent protein governed by the hypoxia‐responsive element in  LLC  cells transplanted into mice,  TGF ‐β‐induced Smad2 phosphorylation was upregulated  in vivo . Our results demonstrate that long‐term exposure to hypoxia might alter responsiveness to  TGF ‐β signaling and affected the malignancy of  LLC  cells.

cancer science

Transforming growth factor‐β signaling enhancement by long‐term exposure to hypoxia in a tumor microenvironment composed of  L ewis lung carcinoma cells