APOBEC3G exerts tumor suppressive effects in human hepatocellular carcinoma.
In this study, we collected 44 hepatitis B virus surface antigen positivity HBsAg (+) tumor and nontumor hepatocellular tissues from hepatocellular carcinoma (HCC) patients during hepatectomy, and quantified the APOBEC3G (A3G) mRNA by using a real-time PCR. Our results showed higher expression of A3G mRNA in the nontumor tissues than in the tumor tissues of the HBsAg (+) HCC patients. To further investigate this phenomenon, we constructed a pLV-A3G vector and transfected it into the human HCC cell line, Hep 3B. The results of an immunofluorescence analysis showed the overexpression of A3G in the cytoplasm. We then evaluated A3G cytotoxicity by using a cell viability assay (MTS assay), the results of which showed that Hep 3B cell viability was 88 and 58% after the transfection of pLV and pLV-A3G, respectively, indicating the growth inhibitory effects of A3G on Hep 3B cells. To further evaluate the tumor suppressive effects of A3G, we used a plastic pipette tip to scratch Hep 3B cells grown on a culture dish (to 70-80% confluence) after transfection with pLV-A3G. Our data indicated a ratio of wound closure of 100% in the control cells and in the pLV-expressing cells, compared with 43% in the pLV-A3G-overexpressing cells, 72 h after the wound scratch, as observed using phase-contrast microscopy. These results indicated that A3G inhibits wound healing in Hep 3B cells. Overall, our results suggest that A3G inhibits the growth of human hepatoma cells.
Departments of aOccupational Therapy bGeneral Surgery cInternal Medicine, Division of Infectious Diseases, E-Da Hospital, I-Shou University dDepartment of Kinesiology, Health and Leisure Studies, National University of Kaohsiung eSchool of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.
Li-Ching
LC
Chang
Ting-Yin
TY
Kuo
Ching-Wen
CW
Liu
Yaw-Sen
YS
Chen
Hsi-Hsun
HH
Lin
Pei-Fung
PF
Wu
D000071480:APOBEC-3G Deaminase; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D045744:Cell Line, Tumor; D002465:Cell Movement; D002470:Cell Survival; D003564:Cytidine Deaminase / Q000235:genetics / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000276:immunology; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D010957:Plasmids; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
D006514:Hepatitis B Surface Antigens; D025521:Tumor Suppressor Proteins; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase