E-cadherin increasing multidrug resistance protein 1 via hypoxia-inducible factor-1α contributes to multicellular resistance in colorectal cancer

Xi Liang, Xueqing Xu, Fengchao Wang, Ni Li, Jianming He

When cancer cells have been cultured as three-dimensional (3D) cultures or in vivo, they decrease sensitivity to anticancer drugs. This is called multicellular resistance, and the mechanism is not fully understood. Here, we report that E-cadherin increasing multidrug resistance protein 1 (MDR1) via hypoxia-inducible factor-1α (HIF-1α) contributes to multicellular resistance in colorectal cancer. The MDR1 protein level was higher in 3D cultures than in monolayer cells. When dispersed cells from 3D cultures were grown as monolayer cells again, the MDR1 protein level decreased to the similar level of cells maintained as monolayer all through. Knockdown of MDR1 significantly decreased multicellular resistance. Knockdown of E-cadherin decreased MDR1 in 3D cultures but did not detectably change MDR1 in monolayer cells. E-cadherin was expressed uniformly in 3D cultures while the MDR1 protein level was higher in the center of 3D cultures than in the peripheral part. Knockdown of E-cadherin decreased E-cadherin uniformly in 3D cultures but mainly decreased MDR1 at the center of 3D cultures. These suggest that knockdown of E-cadherin decreasing MDR1 may be by an indirect mechanism. HIF-1α was remarkably increased in 3D cultures. Knockdown of E-cadherin decreased intercellular junctions, increased intercellular space, and decreased HIF-1α in 3D cultures. Knockdown of HIF-1α decreased MDR1 in 3D cultures. Knockdown of E-cadherin increased β-catenin uniformly in 3D cultures, and knockdown of β-catenin decreased MDR1 what was opposite to knockdown of E-cadherin decreasing MDR1. Our data reveal that knockdown of E-cadherin decreasing MDR1 via HIF-1α is involved in the mechanism of multicellular resistance in colorectal cancer. Though β-catenin is also involved in the mechanism, it does not play a dominant role.