Effects of LncRNA SNHG14 on the Malignant Biological Behaviors of Gastric Cancer Cells by Regulating miR-579-3p/KIF18B Axis
WANG Haiwei, ZHANG Xiaohui*
This study aims to investigate the effects of lncRNA (long non-coding RNA) SNHG14 (small nucleolar RNA host gene 14) on the malignant biological behaviors of GC (gastric cancer) cells by targeting miR-579-3p (microRNA-579-3p)/KIF18B (kinesin family member 18B) axis. RT-qPCR was employed to detect the expression levels of lncRNA SNHG14, miR-579-3p, and KIF18B mRNA in cancer tissues and adjacent normal tissues from 30 GC patients, as well as in the normal human gastric epithelial cell line (GES-1) and GC cell lines (AGS, MGC-803, HGC-27). MGC-803 cells were assigned into sh-NC group, sh-SNHG14 group, sh-SNHG14+anti-miR-NC group, and sh-SNHG14+anti-miR-579-3p group. RT-qPCR was used to detect the expression levels of lncRNA SNHG14, miR-579-3p, and KIF18B mRNA in each group of cells. Cell viability was assessed using the CCK-8 assay, and cell proliferation ability was evaluated by colony formation assay. Flow cytometry was performed to measure cell apoptosis. Wound healing assay was used to detect cell migration ability, and Transwell chamber assay was employed to assess cell invasion ability. Western blot analysis was conducted to determine the protein expression levels of KIF18B, cleaved caspase-3, N-cadherin, and E-cadherin. MGC-803 cells were assigned into miR-NC group, miR-579-3p mimics group, miR-579-3p mimics+pcDNA group, and miR-579-3p mimics+KIF18B group. The expression levels of miR-579-3p and KIF18B mRNA in MGC-803 cells were detected by RT-qPCR. A dual-luciferase reporter assay was performed to validate the targeting relationships between miR-579-3p and lncRNA SNHG14, as well as between miR-579-3p and KIF18B. Silencing lncRNA SNHG14 could downregulate the expression levels of lncRNA SNHG14 and KIF18B mRNA, reduce cell viability, colony number, wound healing rate, and cell invasion number, decrease N-cadherin and KIF18B protein levels, while upregulating miR-579-3p expression, increasing apoptosis rate, and elevating cleaved caspase-3 and E-cadherin protein levels (P<0.05). Silencing lncRNA SNHG14 while downregulating miR-579-3p could upregulate KIF18B mRNA expression levels, increase cell viability, colony number, wound healing rate, cell invasion number, as well as N-cadherin and KIF18B protein expression levels, while reducing miR-579-3p expression levels, cell apoptosis rate, and cleaved caspase-3 and E-cadherin protein expression levels (P<0.05). Overexpression of miR-579-3p could upregulate the expression level of miR-579-3p and downregulate the expression level of KIF18B mRNA (P<0.05); co-transfection of miR-579-3p and KIF18B upregulated KIF18B mRNA expression levels. Dual-luciferase reporter assay confirmed the targeting relationships between miR-579-3p and lncRNA SNHG14, as well as between miR-579-3p and KIF18B. The expression of lncRNA SNHG14 is prominently upregulated in gastric cancer cells. Silencing the expression of lncRNA SNHG14 can target the upregulation of miR-579-3p expression, inhibit the expression of KIF18B, and thus suppress the malignant biological behaviors of gastric cancer cells.
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