[The feasibility and mechanism of recombinant adenovirus Ad - p 14 ARF in gene therapy of liver cancer].
OBJECTIVE
To explore the feasibility and mechanism of recombinant adenovirus Ad-pl4ARF in cancer gene therapy.


METHODS
The proliferation of different liver cancer cells was assessed by morphology and trypan blue assay. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externalization with Annexin V/PI double staining. The expression of related proteins was analyzed by Western bloting. Nude mouse model bearing subcutaneous transplanted BEL7402 tumor was established to study the therapeutic ability of Ad-pl4ARF.


RESULTS
Ad-pl4ARF suppressed cell growth and proliferation, and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-pl4ARF inhibited growth of liver cancer cells ( HepG2, BEL7402) in a dose-dependent manner. Ad-pl4ARF lead to overexpression of Bax and p21, the downstream regulating genes of p53. In the experimental therapy on nude mice bearing subcutaneous transplanted BEL7402 tumor, Ad-pl4ARF suppressed tumor growth significantly.


CONCLUSIONS
pl4ARF is a short gene and with powerful function, which are consistent with the requirements for tumor suppressor genes used in gene therapy. It may play an important role in gene therapy against malignancies in the future.
Song
Hai-feng
HF
State key Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences, PUMC, Beijing, 100021, China.
Qian
Hai-li
HL
Zhang
Xue-yan
XY
Liang
Xiao
X
Fu
Ming
M
Lin
Chen
C
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection; D025803:Tumor Suppressor Protein p14ARF / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D025803:Tumor Suppressor Protein p14ARF; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D016283:Proto-Oncogene Proteins p21(ras)