Experimental study on therapeutic effect of in vivo expression of Cell I-Hep II recombinant polypeptide of fibronectin on murine H22 hepatocellular carcinoma.
OBJECTIVE
To investigate the inhibitory effect of in vivo expression of expressing plasmid pCH510 of recombinant fibronectin polypeptide (CH50) on hepatocellular carcinoma and the improved therapeutic effect of pCH510 in combination with chemotherapeutic agents and Hsp70-H22 hepatocarcinoma antigen peptide on tumor.


METHODS
Mice were inoculated with H22 hepatocarcinoma cells. The chemotactic effect of the expression of plasmid pCH510 on immunocytes was observed after in vivo transfection, tissue slicing and HE staining. Inhibitory effect of transfection with pCH510 on murine tumor originated from different inoculative doses was observed. The inhibitory effect of immediate transfection with pCH510 after chemotherapy on tumor was compared with that of transfection 5 days after chemotherapy. The change of function and amount of mouse peritoneal macrophages and the peripheral blood immunocytes resulted from administration of chemotherapeutic agents were detected. The peptides mixture was prepared from H22 hepatocarcinoma cells. pCH510 + Hsp70-H22 antigen peptides were injected into tumor-bearing mice with or without chemotherapy, to observe the inhibitory effects on tumor.


RESULTS
At the tumor tissue site injected with pCH510, there were a great number of immunocytes which mainly were macrophages, lymphocytes and neutrophils. Transfection of plasmid pCH510 inhibited significantly the murine tumor induced by different inoculative doses. The inhibitory effect was negatively correlated with the inoculative dose. The therapeutic effect was not improved by immediate transfection with pCH510 after chemotherapy, but was significantly improved by transfection with pCH510 5 days after chemotherapy. Chemotherapeutic agent decreased the number of immunocytes and suppressed their activation in vivo. After injection of drug, the amount of immunocytes was the lowest from d 1 to d 3 and returned to normal level on the 10(th) day. Transfection with plasmid pCH510 alone could inhibit tumor induced by the inoculation with 10(4) H22 cells. The tumor originated from the inoculation with 10(5) H22 cells was inhibited by pCH510+Hsp70-H22 antigen peptides and that from the inoculation with 10(6) H22 cells was inhibited by pCH510+Hsp70-H22 antigen peptides in combination with chemotherapeutic agents.


CONCLUSIONS
In vivo expression of pCH510 recruits immune cells, inhibits tumor growth, and enhances the efficacy of chemotherapy. But the proper timing of combining chemotherapy with pCH510 must be taken into great account. The synergism of pCH510 and Hsp70-H22 peptides can improve the efficacy, which could be further enhanced if they are used following chemotherapy. Chemotherapeutic agent + pCH510 + Hsp70-H22 peptides is a promising therapeutic approach of combination treatment of tumor.
Zhang
Gui-Mei
GM
Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
Yang
Yan
Y
Huang
Bo
B
Xiao
Hui
H
Li
Dong
D
Feng
Zuo-Hua
ZH
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002455:Cell Division / Q000187:drug effects; D005353:Fibronectins / Q000494:pharmacology*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D051379:Mice; D010446:Peptide Fragments / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
C478562:CH50 recombinant polypeptide; D005353:Fibronectins; D010446:Peptide Fragments; D011994:Recombinant Proteins