Combined gene therapy of endostatin and interleukin 12 with polyvinylpyrrolidone induces a potent antitumor effect on hepatoma.
OBJECTIVE
To study the antitumor effect of combined gene therapy of endostatin and interleukin 12 (IL-12) with polyvinylpyrrolidone (PVP) on mouse transplanted hepatoma.


METHODS
Mouse endostatin eukaryotic plasmid (pSecES) with a mouse Igkappa signal sequence inside and mouse IL-12 eukaryotic plasmid (pmIL-12) were transfected into BHK-21 cells respectively. Endostatin and IL-12 were assayed by ELISA from the supernant and used to culture endothelial cells and spleen lymphocytes individually. Proliferation of the latter was evaluated by MTT. H22 cells were inoculated into the leg muscle of mouse, which was injected intratumorally with pSecES/PVP, pmIL-12/PVP or pSecES+pmIL-12/PVP repeatedly. Tumor weight, serum endostatin and serum IL-12 were assayed. Tumor infiltrating lymphocytes, tumor microvessel density and apoptosis of tumor cells were also displayed by HE staining, CD31 staining and TUNEL.


RESULTS
Endostatin and IL-12 were secreted after transfection, which could inhibit the proliferation of endothelial cells or promote the proliferation of spleen lymphocytes. Tumor growth was highly inhibited by 91.8% after injection of pSecES+pmIL-12/PVP accompanied by higher serum endostatin and IL-12, more infiltrating lymphocytes, fewer tumor vessels and more apoptosis cells compared with injection of pSecES/PVP, pmIL-12/PVP or vector/PVP.


CONCLUSIONS
Mouse endostatin gene and IL-12 gene can be expressed after intratumoral injection with PVP. Angiogenesis of hepatoma can be inhibited synergisticly, lymphocytes can be activated to infiltrate, and tumor cells are induced to apoptosis. Hepatoma can be highly inhibited or eradiated.
Li
Pei-Yuan
PY
Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Lin
Ju-Sheng
JS
Feng
Zuo-Hua
ZH
He
Yu-Fei
YF
Zhou
He-Jun
HJ
Ma
Xin
X
Cai
Xiao-Kun
XK
Tian
De-An
DA
D000276:Adjuvants, Immunologic / Q000235:genetics*; D020533:Angiogenesis Inhibitors / Q000235:genetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000628:therapy*; D002460:Cell Line; D003131:Combined Modality Therapy; D043169:Endostatins / Q000235:genetics*; D015316:Genetic Therapy; D006801:Humans; D007167:Immunotherapy; D018664:Interleukin-12 / Q000235:genetics*; D008113:Liver Neoplasms / Q000188:drug therapy / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011205:Povidone / Q000627:therapeutic use*
D000276:Adjuvants, Immunologic; D020533:Angiogenesis Inhibitors; D043169:Endostatins; D018664:Interleukin-12; D011205:Povidone