Objective To investigate the impacts of long non-coding RNA（LncRNA） myosin light chain kinase antisense RNA1 （MYLK-AS1） on proliferation，apoptosis，and invasion of gastric cancer cells by regulating the miR-141-3p/stathmin 1（STMN1） axis. Methods HGC27 cells were grouped into negative control（NC） group，si-NC group，si-MYLK-AS1 group，si-MYLK-AS1+inhibitor NC group，and si-MYLK-AS1+miR-141-3p inhibitor group. The relationship among LncRNA MYLK-AS1，miR-141-3p，and STMN1 was determined by the dual-luciferase reporter gene assay；the expression of LncRNA MYLK-AS1 and miR-141-3p in HGC27 cells was measured by qRT-PCR；the CCK-8 method and flow cytometry were usde to determined the proliferation and apoptosis of HGC27 cells，respectively；the numbers of invading and migrating HGC27 cells were counted by Transwell；the protein levels of STMN1，E-cadherin，Vimentin，and N-cadherin in HGC27 cells were measured by Western blot. Results Compared with GES-1 cells，the levels of LncRNA MYLK-AS1 and STMN1 in HGC27 cells were significantly up-regulated（P<0.05），and the level of miR-141-3p was significantly down-regulated（P<0.05）. Compared with the NC group and si-NC group，the si-MYLK-AS1 group showed significant decreases in the optical density at 450 nm，numbers of migrating and invading cells，expression level of LncRNA MYLK-AS1，and protein levels of STMN1，N-cadherin，and Vimentin（P<0.05），as well as significant increases in the apoptosis rate，expression level of miR-141-3p，and protein level of E-cadherin（P<0.05）. However，the low expression of miR-141-3p attenuated the inhibition of HGC27 cell development by silencing LncRNA MYLK-AS1；LncRNA MYLK-AS1 regulated the miR-141-3p/STMN1 axis in a targeted manner. Conclusion Silencing LncRNA MYLK-AS1 may inhibit the expression of STMN1 by up-regulating miR-141-3p，thus affecting the proliferation，apoptosis，and invasion of gastric cancer cells.