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AIM: To determine the inhibitory effect of the vector- generated small interfering RNAs (siRNAs) on the expression of the Bcl-XL gene in established human esophageal cancer cells, and to investigate the effect of the Bcl-XL siRNAs on cell growth and apoptosis in esophageal cancer cells. METHODS: Three siRNA-expressing vectors targeting different sites of the Bcl-XL gene were constructed from pTZ-U6+1 vector. Cultured esophageal cancer cells were transfected with the siRNA-expressing vector (or the control vector) using lipofectamine 2000. Bcl-XL gene expression was determined with semiquantitative RT- PCR assay and Western blotting. Among the three siRNA- expressing vectors, the most highly functional vector and its effect on cell growth and apoptosis in esophageal cancer cells was further analyzed. RESULTS: Of the three siRNA-expressing vectors, siRNA- expressing vector No.1 was the most potent one which suppressed Bcl-XL mRNA production to 32.5% of that in the untreated esophageal cancer cells. Western blotting analysis showed that siRNA-expressing vector No.1 markedly down-regulated the expression of Bcl-XL in human esophageal cancer cells. Treatment of esophageal cancer cells with siRNA-expressing vector No.1 resulted in inhibition of cell growth and induction of apoptosis. CONCLUSION: Down-regulation of Bcl-XL by vector- generated small interfering RNAs can suppress cell growth and induce apoptosis in human esophageal cancer cells.
AIM: To determine the inhibitory effect of the vector-generated small interfering RNAs (siRNAs) on the expression of the Bcl-XL gene in established human esophageal cancer cells, and to investigate the effect of the Bcl-XL siRNAs on cell growth and apoptosis in esophageal cancer cells. METHODS: Three siRNA-expressing vectors targeting different sites of the Bcl-XL gene were constructed from pTZ-U6 + 1 vector. Cultured esophageal cancer cells were transfected with the siRNA- expressing vector (or the control vector) using lipofectamine 2000. Among the three siRNA-expressing vectors, the most highly functional vector and its effect on cell growth and apoptosis in esophageal cancer cells was further analyzed. RESULTS AND TITLE: Bcl-XL gene expression was determined with semiquantitative RT-PCR assay and Western blotting. : Of the three siRNA-expressing vectors, siRNA-expressing vector No.1 was the most potent one which suppressed Bcl-XL mRNA production to 32.5% of that in the untreated esophageal ca ncer cells. Western blotting analysis showed that siRNA-expressing vector No. 1 markedly down-regulated the expression of Bcl-XL in human esophageal cancer cells. Treatment of esophageal cancer cells with siRNA-expressing vector No. 1 resulted in inhibition of cell growth and induction of apoptosis. CONCLUSION: Down-regulation of Bcl-XL by vector-generated small interfering RNAs can suppress cell growth and induce apoptosis in human esophageal cancer cells.