RESUMO
@# Objective: To investigate the expression of long non-coding RNASNHG16 (lncRNASNHG16) in colorectal cancer (CRC) tissues and cells, and to explore the mechanism of its regulation on the expression of mitochondrial glycerol-3-phosphate acyltransferase (GPAM) via sponging miR-128-3p. Methods: Sixty pairs of colorectal cancerous tissues and para-cancerous tissues that resected from CRC patients, who underwent surgery in the Department of Anorectal Surgery, Gansu Provincial People’s Hospital during Jan. 2014 and Jan. 2017, were collected for this study; In addition, CRC cell lines (SW480, SW620, HCT116, Caco-2,DLD-1, HT29) and colonic epithelial cell line CCD841 were also collected for the study. The expression of SNHG16 in collected tissues and cell lines was determined by Real-time quantitative PCR (qPCR), and its correlation to the clinicopathological features of CRC patients was also analyzed. SW480 cells were transfected with miR-128-3p mimic, miR-128-3p inhibitor, and si-SNHG16, respectively, and then the mRNA expressions of miR-128-3p and SNHG16 were detected by qPCR, the protein expression of GPAM was determined by Western blotting, and the cell proliferation, apoptosis and invasion were detected by CCK-8 assay, colony formation assay, cell apoptosis assay and Transwell chamber assay, respectively. The binding between SNHG16 and miR-128-3p was validated with dual luciferase reporter gene assay and RNA Immunoprecipitation assay. For in vivo experiment, mouse model of SW480 cell exnograft was constructed, and the effect of SNHG16 knockdown on the growth of exnograft was observed. Results: SNHG16 was found to highly expressed in human CRC tissues and cell lines (all P<0.01), and SNHG16 expression level was associated with lymph node metastasis, Duke's stage and patients’survival (all P<0.01). Knockdown of SNHG16 significantly inhibited CRC cell proliferation and invasion, and induced apoptosis (all P<0.01); After SNHG16 knockdown, the volume of exnograft was obviously reduced (P<0.05). Dual luciferase reporter gene assay and RNA Immunoprecipitation assay validated the interaction between miR-128-3p and SNHG16, and they were negatively correlated with each other in CRC patients (P<0.01). The SNHG16 regulated the expression of its down-stream gene GPAM via endogenously sponging miR-128-3p. Conclusion: SNHG16 regulates GPAM expression in CRC cells by sponging miR-128-3p, and SNHG16 and miR-128-3p may serve as potential targets for the diagnosis and treatment of CRC.
RESUMO
Objective: To investigate the expression of long non-coding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) in hepatocellular carcinoma (HCC) tissues and cells, and to explore the effects of SNHG16 expression regulation on the proliferation and migration of HCC cells as well as the underlying molecular mechanisms. Methods: The cancer tissues and adjacent tissues were collected from 38 patients with HCC. The real-time fluorescent quantitative PCR was used to detected the expression of SNHG16 in 38 cases of clinical HCC tissue samples and their adjacent tissues, 4 kinds of HCC cell lines and normal hepatocellular cell line. The relationship between SNHG16 expression and the clinicopathological features of HCC patients was analyzed. The SNHG16 overexpression or SNHG16-shRNA recombinant lentivirus was infected into Hep-3B or SK-Hep-1 cells, respectively. The up- or down-regulation of SNHG16 expression in Hep-3B or SK-Hep-1 cells was verified by real-time fluorescent quantitative PCR. The effects of SNHG16 expression regulation on the proliferation and migration of HCC cells were determined by CCK-8 and Transwell chamber experiments, respectively. The expressions of key proteins in Wnt/β-catenin signaling pathway were detected by Western blotting. Xenograft tumor experiment was used to determine the effect of SNHG16 on the tumorigenic ability of HCC cells in nude mice. Results: The expression level of SNHG16 in HCC tissues and HCC cells was significantly higher than that in the adjacent tissues (P < 0.001) and normal hepatocellular cells (P < 0.05), respectively. The expression of SNHG16 in HCC tissues was associated with tumor size (P < 0.01), TNM stage (P < 0.01) and alanine aminotransaminase (ALT) expression level (P < 0.05). After the infection with SNHG16 overexpression recombinant lentivirus, the expression of SNHG16 was significantly up-regulated in Hep-3B cells (P < 0.001), the proliferation and migration of Hep-3B cells were significantly promoted (both P < 0.01), and the expressions of β-catenin and c-myc proteins were up-regulated (both P < 0.01). After the infection with SNHG16-shRNA recombinant lentivirus, the expression of SNHG16 was dramatically downregulated in SK-Hep-1 cells (P < 0.001), the proliferation and migration of SK-Hep-1 cells were significantly inhibited (both P < 0.001), and the expressions of β-catenin (P < 0.05) and c-myc (P < 0.01) proteins were down-regulated. In addition, the tumorigenic ability of Hep- 3B cells with SNHG16 over-expression was significantly enhanced in nude mice (P < 0.01), while the tumorigenic ability of SK-Hep-1 cells with SNHG16 knock-down was significantly weakened in nude mice (P < 0.05). Conclusion: SNHG16 is highly expressed in HCC tissues and cell lines, and may promote the proliferation and migration of HCC cells through Wnt/β-catenin signaling pathway.