Résumé
Objective To investigate the effect of GHET1 on the biological behavior of gallbladder cancer cells and the regulatory mechanism of GHET1 on miR-27b. Methods The expression of GHET1 and miR-27b in 50 samples of gallbladder cancer was detected by real-time quantitative PCR. The si-NC vector, si-GHET1 vector, miR-27b inhibitor, and si-GHET1 vector+miR-27b inhibitor were transfected into SGC-996 cells and set as the control group, GHET1 interference group, miR-27b interference group, and GHET1+miR-27b interference group. Cell proliferation, apoptosis, and metastasis in each group were detected by MTT, flow cytometry, and Transwell assays. The regulatory effect of GHET1 on miR-27b was validated by luciferase reporter gene assay. Results GHET1 expression was higher in cancer tissues than that in paracancerous ones. miR-27b expression was lower in cancer tissues than that in paracancerous tissues. GHET1 was negatively correlated with miR-27b expression (P<0.05), and GHET1 expression was associated with TNM staging and lymph node metastasis (P<0.05). High GHET1 expression was associated with poor prognosis of patients with gallbladder cancer (P<0.05). Compared with the control group, the GHET1 interference group showed decreased cell-proliferation ability, increased apoptosis rate, and reduced number of cell metastasis. The miR-27b interference group showed increased cell-proliferation ability, decreased apoptosis rate, and increased number of cell metastasis (P<0.05). Compared with the GHET1 interference group, the GHET1+miR-27b interference group showed increased cell-proliferation ability, decreased apoptosis rate, and increased number of cell metastasis (P<0.05). GHET1 inhibited miR-27b expression by acting as a sponge of miR-27b. Conclusion GHET1 promotes the proliferation and metastasis and inhibits the apoptosis of gallbladder cancer cells by targeting miR-27b, suggesting that GHET1/miR-27b axis plays a role in gallbladder cancer progression.
Résumé
Objective To investigate the expression of long-chain non-coding RNA gastric cancer high expression transcription factor 1 (GHET1) in hepatocellular carcinoma (HCC) and the correlation with prognosis,cell proliferation,migration and invasion.Methods 20 HCC patients who underwent surgery from Fuzhou General Hospital of Chinese People's Liberation Army from March to May 2016 were included.The HCC tissue and adjacent normal tissue of 182 patients from June 2012 to December 2013 were retrospectively collected.According to the median value of GHET1 expression,it was divided into GHET1 high expression group and low expression group,91 cases each.Huh7 and HepG2 cells were divided into:blank control group (Con) with serum-free medium,siRNA-GHET1 group transfected with siRNA-GHET1,and negative control group (siRNA-NC) transfected with negative control sequence.The expression of GHET1 was detected by real-time fluorescence quantitative polymerase chain reaction,and the effect of GHET1 on HCC cells was analyzed by CCK-8,Transwell assay and Western blot.Results Compared with adjacent normal tissue,the relative expression of GHET1 mRNA in HCC tissues was significantly increased.Compared with LO2 cells,the mRNA expression of GHET1 in Huh7 and HepG2 cells was higher (P＜0.05).The GHET1 high expression group had tumor＞5 cm,vascular invasion,AFP＞400 μg/L,Edmonson grade Ⅰ,and the tumor-free ratio was lower in the expression group (P＜0.05).Survival analysis showed that HCC patients with high GHET1 expression had a poorer prognosis than patients with low expression.Multivariate Cox regression analysis showed that high expressed GHET1,vascular invasion (HR=2.067,95％ CI:1.350 to 3.162),and without tumor capsule are independent predictors of recurrence in HCC patients.After transfection with Huh7 and HepG2 cells,the proliferation of siRNA-GHET1 group was significantly decreased comparing with Con and siRNA-NC groups.Compared with siRNA-NC group,the migration and invasion ability of siRNA-GHET1 group decreased,and E-cadherin expression increased.The expression of fibronectin and vimentin decreased,and the difference was statistically significant (P＜0.05).Conclusions The expression of GHET1 in HCC tissue is higher comparing with normal tissue,which increases the proliferation,migration and invasion of hepatoma cells.It is an independent predictor of prognosis in HCC patients and a potential target for clinical treatment.