Cancer metastasis, which is prevalent in malignant tumors, is present in a variety of cases depending on the primary tumor and metastatic site. The cancer metastasis is affected by various factors that surround and constitute a tumor microenvironment. One of the several factors, oxygen tension, can affect cancer cells and induce changes in many ways, including motility, directionality, and viability. In particular, the oxygen tension gradient is formed within a tumor cluster and oxygen is lower toward the center of the cluster from the perivascular area. The simple and efficient designing of the tumor microenvironment using microfluidic devices enables the simplified and robust platform of the complexin vivomicroenvironment while observing a clear cause-and-effect between the properties of cancer cells under oxygen tension. Here, a microfluidic device with five channels including a gel channel, media channels, and gas channels is designed. MDA-MB-231cells are seeded in the microfluidic device with hydrogel to simulate their three-dimensional movement in the body. The motility and directionality of the cancer cells under the normoxic and oxygen tension gradient conditions are compared. Also, the viability of the cancer cells is analyzed for each condition when anticancer drugs are applied. Unlike the normoxic condition, under the oxygen tension gradient, cancer cells showed directionality toward higher oxygen tension and decreased viability against the certain anticancer drug. The simplified design of the tumor microenvironment through microfluidic devices enables comprehension of the response of cancer cells to varying oxygen tensions and cancer drugs in the hypoxic tumor microenvironment.