Eukaryotic elongation factor-1α 2 knockdown inhibits hepatocarcinogenesis by suppressing PI3K/Akt/NF-κB signaling.
OBJECTIVE
To assess the impact of eukaryotic elongation factor 1 alpha 2 (eEF1A2) on hepatocellular carcinoma (HCC) cell proliferation, apoptosis, migration and invasion, and determine the underlying mechanisms.


METHODS
eEF1A2 levels were detected in 62 HCC tissue samples and paired pericarcinomatous specimens, and the human HCC cell lines SK-HEP-1, HepG2 and BEF-7402, by real-time PCR and immunohistochemistry. Experimental groups included eEF1A2 silencing in BEL-7402 cells with lentivirus eEF1A2-shRNA (KD group) and eEF1A2 overexpression in SK-HEP-1 cells with eEF1A2 plasmid (OE group). Non-transfected cells (control group) and lentivirus-based empty vector transfected cells (NC group) were considered control groups. Cell proliferation (MTT and colony formation assays), apoptosis (Annexin V-APC assay), cell cycle (DNA ploidy assay), and migration and invasion (Transwell assays) were assessed. Protein levels of PI3K/Akt/NF-κB signaling effectors were evaluated by Western blot.


RESULTS
eEF1A2 mRNA and protein levels were significantly higher in HCC cancer tissue samples than in paired pericarcinomatous and normal specimens. SK-HEP-1 cells showed lower eEF1A2 mRNA levels; HepG2 and BEL-7402 cells showed higher eEF1A2 mRNA levels, with BEL-7402 cells displaying the highest amount. Efficient eEF1A2 silencing resulted in reduced cell proliferation, migration and invasion, increased apoptosis, and induced cell cycle arrest. The PI3K/Akt/NF-κB signaling pathway was notably inhibited. Inversely, eEF1A2 overexpression resulted in promoted cell proliferation, migration and invasion.


CONCLUSIONS
eEF1A2, highly expressed in HCC, is a potential oncogene. Its silencing significantly decreases HCC tumorigenesis, likely by inhibiting PI3K/Akt/NF-κB signaling.
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Fu-Nan
FN
Qiu
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Yi
Y
Huang
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Dun-Yan
DY
Chen
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Feng
F
Li
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Yan-An
YA
Wu
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Wen-Bing
WB
Wu
Fu-Nan Qiu, Yi Huang, Dun-Yan Chen, Feng Li, Yan-An Wu, Wen-Bing Wu, Xiao-Li Huang, Provincial Clinical College, Fujian Medical University, Fuzhou 350001, Fujian Province, China.
Xiao-Li
XL
Huang
D000293:Adolescent; D000328:Adult; D000368:Aged; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D016022:Case-Control Studies; D059447:Cell Cycle Checkpoints; D002465:Cell Movement; D049109:Cell Proliferation; D015536:Down-Regulation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D055815:Young Adult
C503234:EEF1A2 protein, human; D016328:NF-kappa B; D020648:Peptide Elongation Factor 1; D012333:RNA, Messenger; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt
Carcinogenesis; Eukaryotic elongation factor 1 alpha 2; Hepatocellular carcinoma; PI3K/Akt/NF-κB signaling pathway; Proliferation