Transfection of thymidine phosphorylase cDNA to human hepatocellular carcinoma cells enhances sensitivity to fluoropyrimidine but augments endothelial cell migration.

Abstract

PURPOSE:

To investigate the effects on sensitivity to fluoropyrimidine and endothelial cell (EC) migration by transfection with thymidine phosphorylase (TP) cDNA to a hepatocellular carcinoma (HCC) cell line SMMC-7721.

METHODS:

SMMC-7721 was transfected with pcDNA3.1/zeo (+) with human TP cDNA. TP mRNA expression was determined by RT-PCR. Sensitivity to fluoropyrimidine was determined by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Induction of EC migration was detected by Boyden chamber assay.

RESULTS:

The construction of pcDNA3.1/zeo(+)-TP was verified by digestion with restriction endonuclease Apa1. When comparison was made between SMMC-7721 cell clone transfected with pcDNA3.1/zeo(+)-TP (S-TP) and control clone transfected with pcDNA3.1/zeo(+) (S-vector), we found that TP mRNA expression level was much higher in S-TP, being 2.09+/-0.16 vs 0.48+/-0.06 in S-vector (P < 0.01), sensitivity to 5'-deoxy-5-fluorouridine (5'-dFUrd, a prodrug of 5-fluorouracil) in S-TP was significantly enhanced compared with that in S-vector (IC(50); 56.81+/-9.85 micromol/l vs 162.25+/-11.03 micromol/l, P < 0.01), and the culture medium of S-TP possessed more potential to induce EC migration than that of S-vector (the number of ECs appearing on the outer surfaces of the membrane was 275+/-29 vs 122+/-35 per field, P < 0.01).

CONCLUSION:

Sensitivity to 5'-dFUrd could be enhanced by transfection with TP cDNA for SMMC-7721 cells. However, EC migration was also promoted at the same time. Therefore, transfection with TP alone might have no potential to enhance anti-tumoral effects of fluoropyrimidine in HCC.

• Authors: Jian Zhou, Yong-Sheng Xiao, Zhao-You Tang, Jia Fan, Zhi-Quan Wu, Yan Zhao, Qiong Xue, Zao-Zhuo Shen, Yin-Kun Liu, Sheng-Long Ye
• Journal: Journal of cancer research and clinical oncology (J Cancer Res Clin Oncol) Vol. 131 Issue 8 Pg. 547-51 (Aug 2005) ISSN: 0171-5216 [Print] Germany
• PMID: 15864645 (Publication Type: Journal Article)

Chemical References

• Antigens, Neoplasm
• Antimetabolites, Antineoplastic
• DNA, Complementary
• DNA-Binding Proteins
• Floxuridine
• Thymidine Phosphorylase
• DNA Topoisomerases, Type II
• doxifluridine

Topics

• Antigens, Neoplasm (analysis)
• Antimetabolites, Antineoplastic (pharmacology)
• Carcinoma, Hepatocellular (drug therapy, enzymology, pathology)
• Cell Movement (drug effects)
• DNA Topoisomerases, Type II (analysis)
• DNA, Complementary (metabolism)
• DNA-Binding Proteins (analysis)
• Disease-Free Survival
• Floxuridine (pharmacology)
• Humans
• Immunohistochemistry
• In Situ Hybridization, Fluorescence
• Liver Neoplasms (drug therapy, enzymology, pathology)
• Reverse Transcriptase Polymerase Chain Reaction
• Survival Analysis
• Thymidine Phosphorylase (genetics)
• Transfection