objective To observe the effect of Na+/H+ exchanger 1 (NHE1)-regulated intracellular pH on the invasion and migration of gastric cancer cell line SGC-7901. Methods The gastric cancer cell line SGC -7901 was treated with the NHE1 specific inhibitor 5-(N-ethyl-N-isopropyl) Amiloride (EIPA), the intracellular pH of SGC-7901 was measured using confocal fluorescence microscope. The migration of SGC-7901 cells was examined by scratch test. The invasion of SGC-7901 cells was examined by transwell chamber invasive test. Results The intracellular pH in the control group was (7.2 ± 0.12), it significantly reduced to (7.01 ± 0.23) in the 5-μmol EIPA group. EIPA of different concentrations (5 μmol, 10 μmol and 20 μmol) could inhibit the invasion and migration of SGC-7901 cells in a dose-dependent manner. Conclusions NHE1 plays an important role in the regulation of intracellular pH. Blockage of NHE1 could restrain the invasion and migration of gastric cancer cells.