2013Molecular Medicine ReportsOpen access

Effects of cyclin E gene silencing on the proliferation of esophageal cancer cell lines, EC9706, Eca109 and KYSE30

Na Wang, Min Li, Wenqiao Zang, Yunyun Ma, Yuanyuan Wang, Guoqiang Zhao

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Abstract

In order to observe the effects of cyclin E gene silencing by small interfering RNA (siRNA) on the growth, proliferation, invasion and apoptosis of esophageal cancer cell lines, including EC9706, Eca109 and KYSE30, siRNA vectors targeting cyclin E gene were constructed and then transfected into the EC9706, Eca109 and KYSE30 human esophageal cancer cell lines. Cyclin E mRNA and protein expression were determined by RT-PCR and western blotting. Cell proliferation and clonality were detected using a CCK-8 test and soft agar colony formation assay. Cell cycle distribution, apoptosis and invasion of EC9706, Eca109 and KYSE30 cells were evaluated with flow cytometry and a transwell culture system. After siRNA vectors targeting the cyclin E gene were transfected into EC9706, Eca109 and KYSE30 cell lines, compared with blank and negative control groups, the expression of cyclin E mRNA and protein (P<0.01), colony-forming units and the number of cells penetrating the transwell membrane (P<0.05) were significantly decreased, the cells in the S and G2/M phase were reduced, the cells in the G0/G1 phase were increased and the apoptosis rate was increased (P<0.01) in the experimental groups. Cyclin E gene silencing effectively inhibits growth, proliferation and invasion of esophageal cancer cells.

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In order to observe the effects of cyclin E gene silencing by small interfering RNA (siRNA) on the growth, proliferation, invasion and apoptosis of esophageal cancer cell lines, including EC9706, Eca109 and KYSE30, siRNA vectors targeting cyclin E gene were constructed and then transfected into the EC9706, Eca109 and KYSE30 human esophageal cancer cell lines. Cyclin E mRNA and protein expression were determined by RT-PCR and western blotting. Cell proliferation and clonality were detected using a CCK-8 test and soft agar colony formation assay. Cell cycle distribution, apoptosis and invasion of EC9706, Eca109 and KYSE30 cells were evaluated with flow cytometry and a transwell culture system. After siRNA vectors targeting the cyclin E gene were transfected into EC9706, Eca109 and KYSE30 cell lines, compared with blank and negative control groups, the expression of cyclin E mRNA and protein (P<0.01), colony-forming units and the number of cells penetrating the transwell membrane (P<0.05) were significantly decreased, the cells in the S and G2/M phase were reduced, the cells in the G0/G1 phase were increased and the apoptosis rate was increased (P<0.01) in the experimental groups. Cyclin E gene silencing effectively inhibits growth, proliferation and invasion of esophageal cancer cells.

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Available abstract

In order to observe the effects of cyclin E gene silencing by small interfering RNA (siRNA) on the growth, proliferation, invasion and apoptosis of esophageal cancer cell lines, including EC9706, Eca109 and KYSE30, siRNA vectors targeting cyclin E gene were constructed and then transfected into the EC9706, Eca109 and KYSE30 human esophageal cancer cell lines. Cyclin E mRNA and protein expression were determined by RT-PCR and western blotting. Cell proliferation and clonality were detected using a CCK-8 test and soft agar colony formation assay. Cell cycle distribution, apoptosis and invasion of EC9706, Eca109 and KYSE30 cells were evaluated with flow cytometry and a transwell culture system. After siRNA vectors targeting the cyclin E gene were transfected into EC9706, Eca109 and KYSE30 cell lines, compared with blank and negative control groups, the expression of cyclin E mRNA and protein (P<0.01), colony-forming units and the number of cells penetrating the transwell membrane (P<0.05) were significantly decreased, the cells in the S and G2/M phase were reduced, the cells in the G0/G1 phase were increased and the apoptosis rate was increased (P<0.01) in the experimental groups. Cyclin E gene silencing effectively inhibits growth, proliferation and invasion of esophageal cancer cells.

Key concepts: Cell cycle, Transfection, Molecular biology, Biology, Gene silencing, Cell growth, Cyclin E, Cyclin B1

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Effects of cyclin E gene silencing on the proliferation of esophageal cancer cell lines, EC9706, Eca109 and KYSE30 — Research Paper | ScholarLens