2008Zhōnghuá xiāohuà wàikē zázhì/Zhonghua xiaohua waike zazhiRequires access

BC047440 gene silencing inhibits the proliferation of HepG2 cells

Xiaobing Huang, Jing Li, Ping Liang, Lu Zheng, Shicheng Liu, Keqiang Han

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Abstract

Objective To investigate whether the proliferation of HepG2 ceils is influenced by interfering BC047440 gene with small hairpin RNA (shRNA) expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA (shRNA1 and shRNA2) were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells. Key words: BC047440 gene;  RNA interference;  Liver cancer

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What this paper is about

Objective To investigate whether the proliferation of HepG2 ceils is influenced by interfering BC047440 gene with small hairpin RNA (shRNA) expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA (shRNA1 and shRNA2) were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells. Key words: BC047440 gene;  RNA interference;  Liver cancer

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Available abstract

Objective To investigate whether the proliferation of HepG2 ceils is influenced by interfering BC047440 gene with small hairpin RNA (shRNA) expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA (shRNA1 and shRNA2) were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells. Key words: BC047440 gene;  RNA interference;  Liver cancer

Key concepts: Small hairpin RNA, Molecular biology, Transfection, RNA interference, Plasmid, Cell growth, Gene silencing, Cell cycle

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