[A study on the apoptosis of hepatoma cells synergistically induced by plasmid-mediated anti-angiogenesis and immunopotentiation therapy].
OBJECTIVE
To study the effect and mechanism of apoptosis of implanted hepatoma cells in mice induced by eukaryotic plasmid-mediated anti-angiogenesis and immunopotentiation therapy.


METHODS
Mouse endostatin eukaryotic plasmid (pSecES) and mouse IL-12 (interleukin 12) eukaryotic plasmid (pmIL-12) were extracted and purified from E.coli. H22 hepatoma cells were inoculated into the leg muscles of 32 mice. The mice were divided into four groups with pSecES, pmIL-12, pSecES+pmIL-12 and pcDNA3.1 naked plasmid DNA injected into the tumor cell implantation sites of each group. Tumor formation and their weights were evaluated. Microvessel density, numbers of the infiltrating lymphocytes in the tumors and the apoptosis of tumor cells were assayed by microscopical examination of the CD31 and HE stained slides of the tumors and TUNEL assay.


RESULTS
The tumors of those with pSecES or pmIL-12 injections grew slower and with less microvessel density, more lymphocyte infiltration and with more apoptosis tumor cells compared with those with pcDNA3.1 injections. There was much more tumor cell apoptosis in the pSecES+pmIL-12 group (19.9+/-5.5 per 400x microscope field, P less than 0.05) than that in any other single plasmid injection group (400x microscopic field: pSecES 11.3+/-4.1, pmIL-12 14.6+/-3.2, pcDNA3.1 1.4+/-1.3).


CONCLUSIONS
Tumor cell apoptosis of the implanted hepatoma in mice can be induced by eukaryotic plasmid-mediated anti-angiogenesis and immunotherapy through inhibiting tumor angiogenesis and promoting tumor lymphocyte infiltration, by which the growth of the implanted hepatoma was inhibited.
Li
Pei-Yuan
PY
Division of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Huang
Huan-Jun
HJ
Zhang
Qiong
Q
D000818:Animals; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D045744:Cell Line, Tumor; D043169:Endostatins / Q000235:genetics*; D007167:Immunotherapy; D018664:Interleukin-12 / Q000235:genetics*; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009368:Neoplasm Transplantation; D010957:Plasmids
D043169:Endostatins; D018664:Interleukin-12