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The aim of this study was to characterize the involvement of long noncoding RNA HOTAIR in gastric cancer development. Measurement of HOTAIR and miRNA-206 expression by in situ hybridization (ISH) and analyzed for the correlation between HOTAIR and miRNA-206 in gastric cancer tissues. To evaluate the effects of HOTAIR in gastric cancer, MTT assay, flow cytometry, transwell, and wound healing assays were applied. To explain the mechanism behind HOTAIR's involvement, the expression of proteins related to it was also measured by Western blo