MiR-335 Directly, While MiR-34a Indirectly Modulate Survivin Expression and Regulate Growth, Apoptosis, and Invasion of Gastric Cancer Cells

miR-335 and miR-34a are two microRNAs (miRNAs) usually downregulated in gastric cancer (GC). But, their exact regulative roles were not fully elucidated. In this study, we studied the association between miR-335 and/or miR-34a expression and overall survival of GC patients and explored the regulative role of miR-335 and -34a over survivin expression and GC cell growth and invasion. Fifty patients with GC were regularly followed up from 2011 to 2015. miRNA microarray was used to examine the expression trend of miRNAs in eight tumor tissue samples and adjacent normal tissue samples. The possible binding site between miR-335 and survivin messenger RNA (mRNA) was predicted using online database and verified using qRT-PCR, Western blot, and dual luciferase assay. The regulative role of miR-335 and miR-34a over GC cell growth, apoptosis, and invasion was studied using Cell Counting Kit-8 (CCK-8) assay, flow cytometry, and transwell assay, respectively. Among the GC patients, low miR-335 or miR-34a expression is associated with higher clinical stage and lymph node metastasis. Patients with low miR-335 or miR-34a had poor overall survival, while those with combined low miR-335 and miR-34a expression had even poorer overall survival. miR-335 can directly regulate survivin expression through binding to the 3'UTR, while miR-34a has indirect modulating effect. Both miR-335 and miR-34a could inhibit cell proliferation and invasion and enhance cell apoptosis. But, these effects are largely abrogated by overexpression of survivin without 3'UTR. Therefore, besides the targets identified in previous studies, miR-335 and miR-34a can also regulate GC cell growth, apoptosis, and invasion at least partly through survivin.