TIMP-3 gene transfection suppresses invasive and metastatic capacity of human hepatocarcinoma cell line HCC-7721.
BACKGROUND
Tissue inhibitor of metalloproteinases (TIMPs) can restrain the tumor growth by their protein property and matrix metalloproteinases (MMPs) inhibition. There are currently four known TIMP family members-TIMP-1, 2, 3, 4. We determined whether increasing levels of TIMP-3 expression could suppress the malignant phenotype of human hepatocarcinoma cell line HCC-7721.


METHODS
A recombinant expression vector, which contained full-length cDNA of human TIMP-3, was constructed and transfected into the human hepatocarcinoma cell line HCC-7721 by the lipofectamine technique. In vitro and in vivo tests such as Western blotting, immunohistochemistry as well as xenografting in nude mice were used to analyze expression levels of TIMP and MMP, and changes in malignant phenotype after the gene transfection.


RESULTS
TIMP-3 expression in TIMP-3 gene-transfected HCC-7721 cells was upregulated as assessed by Western blotting. The ability of in vitro invasion through a Boyden chamber was significantly decreased compared to controls. Following subcutaneous injection into nude mice, the TIMP-3 transfected cells suppressed primary tumor growth, as characterized by reduced tumor weight, size and microvasculature as well as maintaining the extracellular matrix.


CONCLUSIONS
The results suggest that upregulation of TIMP-3 expression in HCC-7721 cells inhibits invasion capacity in vitro as well as tumorigenic and metastatic potential in nude mice.
Zhang
Hong
H
Department of General Surgery, Second Hospital of Jilin University, Changchun 130041, China. zhanghong519@hotmail.com
Wang
Yue-Shu
YS
Han
Gang
G
Shi
Ying
Y
D000818:Animals; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000235:genetics* / Q000556:secondary; D045744:Cell Line, Tumor; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D012334:RNA, Neoplasm / Q000235:genetics*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D012334:RNA, Neoplasm; C088390:TIMP3 protein, human; D019717:Tissue Inhibitor of Metalloproteinase-3