Overexpression of Bax up-regulates the sensitivity of HCC-9204 cell apoptosis induced by adriamycin.
OBJECTIVE
To investigate whether Bax could regulate the sensitivity of human HCC-9204 cells to adriamycin.


METHODS
Overexpression of Bax was induced through inducible system and MT-II regulatory system, with addition of ZnSO(4) (100 micro mol L(-1)) as external inducer. Stable transfecting inducible expression vector containing Bax gene was performed. Apoptotic cells were measured by morphological criteria, and detection of apopto tic DNA fragmentation by TUNEL assay and flow cytometry. The ability of Bax to lower clonogenic cell survival rate was studied by colony-forming assay, while decrease of cell viability was assessed by MTT assay.


RESULTS
HC-9204 cells treated with adriamycin (20 micro mol/L) showed extensive cell death with nucleus fragmentation detected by TUNEL assay. FACS analysis showed a significant sub-G(1) peak and apoptosis in 31% cells 24 hr after treatment. Bax was able to significantly decrease clonogenic survival rate in adriamycin treatment group, showing time dependence. Bax could selectively sensitize HCC-9204 cells to cell death induced by DNA damaging agent-adriamycin.


CONCLUSIONS
Overexpression of Bax is able to sensitize HCC-9204 cell apoptosis induced by a driamycin.
Zheng
Jianyong
J
Department of Gastrointestinal Surgery, Xijing Hospital, Fourth Military Medical University, Shanxi 710032, China.
Li
Jiang
J
Wang
Wenliang
W
Wang
Weizhong
W
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D004317:Doxorubicin / Q000494:pharmacology*; D015870:Gene Expression; D006801:Humans; D008113:Liver Neoplasms; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D051028:bcl-2-Associated X Protein
D000970:Antineoplastic Agents; C494826:BAX protein, human; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D004317:Doxorubicin