Abstract
To evaluate the effect of antisense vascular endothelial growth factor (VEGF) RNA (PCMV-FGEV) transfection on the profile of hepatocellular carcinoma (HCC) SMMC-7721 cells in vitro and in vivo. SMMC-7721 cells were transfected with PCMV-FGEV antisense, PCMV-VEGF sense and empty vector plasmid encapsulated by lipofectamine as antisense group, sense group and control group respectively. The positive cell clones were selected with G418. The stable transfection and expression of VEGF in the cells were determined by RT-PCR and immunohistochemistry. Cell proliferation was observed by MTT assay. FACS analysis was used to determine the effect of PCMV-FGEV transfection on cell apoptosis. The growth of transfected cells in vivo was also observed in nude mice. VEGF expression was reduced in SMMC-7721 transfected with PCMV-FGEV, which was confirmed by RT-PCR and immunohistochemistry. No effect of PCMV-FGEV transfection was found on cell proliferation and cell apoptosis of SMMC-7721 in vitro. The growth of cells transfected with PCMV-FGEV was slow in nude mice and accompanied with obvious apoptosis. The latent time of tumors in the antisense group was 25.0 +/- 1.8 d, which was longer than that in sense and control groups (F = 19.455, P < 0.01). The average tumor weight in antisense group (0.96 g +/- 0.28 g) was the smallest among the three groups (F = 21.501, P < 0.01). The expression of VEGF can be inhibited by antisense PCMV-FGEV. Antisense PCMV-FGEV has no effect on cell proliferation and apoptosis of SMMC-7721 in vitro but can inhibit tumor growth and induce cell apoptosis in vivo.
MeSH Terms
Animals
Apoptosis/drug effects
Carcinoma, Hepatocellular/chemistry,drug therapy,pathology,ultrastructure
Cell Line, Tumor
Cell Proliferation/drug effects
Factor VIII/analysis,genetics
Female
Flow Cytometry
Gene Expression Regulation, Neoplastic/drug effects
Humans
Liver/drug effects,pathology
Liver Neoplasms/chemistry,drug therapy,pathology,ultrastructure
Mice
Mice, Inbred BALB C
Mice, Nude
RNA, Antisense/pharmacology,therapeutic use
Reverse Transcriptase Polymerase Chain Reaction
Transfection
Vascular Endothelial Growth Factor A/analysis,genetics
Chemicals
RNA, Antisense
Vascular Endothelial Growth Factor A
Factor VIII
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hao Ji-Hui
Department of Abdominal Surgery, Tianjin Cancer Hospital, Hexi District, Tianjin 300060, China. jihuihao@yahoo.com
Yu Ming
Li Hui-Kai
Shi Yu-Rong
Li Qiang
Hao Xi-Shan
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