Objective: To study the effects of eukaryotic translation elongation factor 1A1 (eEF1A1) on the cell cycle and proliferation of hepatocellular carcinoma (HCC) cells, and to explore the molecular mechanism. Methods: The expression of eEF1A1 protein in 101 HCC tissues and their adjacent tissues were detected by immunohistochemistry, and the correlations between the expression of eEF1A1 protein and the clinicopathologic parameters of HCC patients was further analyzed. The expressions of eEF1A1 mRNA and protein in HCC cells (including Huh7, SMMC7721, MHCC97H and Hep3B) and the normal liver cells HL-7702 were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The specific shRNA-1/2 targeting eEF1A1 gene (eEF1A1-shRNA-1/2) and the overexpression plasmid pcDNA3.1- eEF1A1 were respectively transfected into Huh7 and SMMC7721 cells, and the alteration of eEF1A1 expression was verified by real-time fluorescent quantitative PCR and Western blotting, respectively. Then the proliferation and cell cycle of HCC cells were detected by EdU and FCM assays, respectively. In addition, the expressions of cyclin D2 and the signal transducer and activator of transcription 1 (STAT1)-related pathway proteins in Huh7 cells transfected with eEF1A1-shRNA-1/2 and in SMMC7721 cells transfected with pcDNA3.1- eEF1A1 were detected by Western blotting. Results: The expression level of eEF1A1 protein in HCC tissues was markedly higher than that in the adjacent tissues (P < 0.001). The expression levels of eEF1A1 mRNA and protein in four HCC cell lines were significantly higher than those in normal live cells (all P < 0.001). The high expression of eEF1A1 was closely related to tumor size and TNM stage in HCC patients (P < 0.01, P < 0.05). After transfection of eEF1A1-shRNA-1/2 into Huh7 cells, the expression levels of eEF1A1 mRNA and protein were significantly decreased (both P < 0.01), the proportion of G1 phase-cells increased significantly (P < 0.01), and the proliferation activity of Huh7 cells was significantly reduced (P < 0.01). After transfection of pcDNA3.1-eEF1A1 into SMMC7721 cells, the expression levels of eEF1A1 mRNA and protein were significantly increased (both P < 0.01), the proportion of G1 phase-cells was significantly reduced (P < 0.05), and the proliferation activity of SMMC7721 cells increased significantly (P < 0.05). In addition, the expression levels of cyclin D2, total and nuclear STAT1 proteins were decreased (all P < 0.05) in Huh7 cells with the downregulation of eEF1A1 expression. After eEF1A1 overexpression, the expression levels of cyclin D2, total and nuclear STAT1 proteins were increased (all P < 0.05) in SMMC7721 cells. However, the expression levels of cyclin D2, total and nuclear STAT1 proteins were unchanged (all P > 0.05) in SMMC7721 cells after eEF1A1 overexpression and treatment with STAT1 inhibitor fludarabine. Conclusion: eEF1A1 regulates the expression of cyclin D2 through STAT1 pathway, thereby promoting the cell cycle progression and proliferation of primary HCC cells.

Objective To investigate the effect of safranal on neurologic functions and histopathologic changes after spinal cord injury (SCI)and the related molecular mechanisms.Methods We randomly assigned 36 rats into six groups:control group,injury group and four treatment groups (namely,A,B,C,and D).The Basso Beattie Bresnahan locomotor rating scale (BBB)and HE staining were applied to evaluate the neuroprotective effect and determine the most effective dosage.Another 60 rats were randomly and evenly assigned to three groups:control group,injury group and treatment group.Nissl staining,TUNEL staining and electron microscopy were used to analyze histopathological changes;RT-PCR,immunohistopathological staining,ELISA,and Western blot were used to detect the expressions of apoptosis-related proteins (Bax and Bcl-2 ),inflammation-related factors (IL-1β, IL-10,TNF-αand P38MAPK),and edema-related factor (APQ-4).Results The optimal dosage for safranal was 100 mg/kg.Neurocyte structure was found more distinct in treatment group than in injury group.In addition,we detected a smaller number of apoptotic neurocyte (26.37±1.54 vs.35.94±1.62,P=0.000),decreased Bax (P=0.000)and APQ-4[(359.55±16.12)% vs.(124.53±20.35)%,P=0.000]expressions,increased Bcl-2 (P=0.036)expression,and obviously lowered P38MAPK [(300.30±33.26)% vs.(132.54±10.21)%,P=0.000]expression. Conclusion Safranal exerts its neuroprotective function through anti-apoptosis,anti-inflammation and anti-edema in the rat model of spinal cord injury.

OBJECTIVETo explore the role of eukaryotic translation elongation factor 1A1 (eEF1A1) in regulating the invasion and metastasis of hepatocellular carcinoma (HCC) cells and the possible mechanism.

METHODSqRT-PCR and Western blotting were used to detect the mRNA and protein expression of eEF1A1 and NOB1 in different HCC cell lines and normal liver cells. The invasion and migration abilities of HCC cells with eEF1A1 knockdown or overexpression were examined using Transwell chamber assay and RTCA assay, and the changes in NOB1 mRNA and protein expressions in the cells were detected. The effects of increasing NOB1 expression in HCCLM3-sheEF1A1 cells and decreasing NOB1 expression in eEF1A1-overexpressing MHCC97h cells on eEF1A1 expression and cell invasion and migration abilities were analyzed using Western blotting, Transwell chamber assay and RTCA assay.

RESULTSThe expressions of eEF1A1 and NOB1 were significantly increased in positive correlation in HCC cells as compared with normal hepatocytes. Knockdown of eEF1A1 significantly decreased the invasion and migration of HCC cells and reduced the mRNA and protein expression of NOB1 ( < 0.01). Overexpression of eEF1A1 significantly enhanced invasion and migration of HCC cells and increased NOB1 mRNA and protein expressions ( < 0.01). Increasing NOB1 expression in HCCLM3-sheEF1A1 cells led to the restoration of NOB1 expression and cell invasion and migration abilities ( < 0.01), whereas decreasing NOB1 in MHCC97h-eEF1A1 cells resulted in inhibition of NOB1 expression and cell invasion and migration ( < 0.01).

CONCLUSIONSeEF1A1 positively regulates the expression of NOB1 to promote the invasion and migration of HCC cells .