Effects to HeLa Cells of the Inhibition of Survivin by Antisense RNA.
Zhu-Zhong Mei, Yan Dong, Zhixian Sun
Abstract
Zhu-Zhong Mei, Yan Dong, Zhixian Sun
Abstract
To study the function of survivin in cancer cells, survivin cDNA was amplified from HeLa cells by RT-PCR. Then the coding fragment was subcloned into an inducible eukaryotic expression plasmid pHC in reverse direction.The obtained plasmid pHSC was transfected into HeLa cells by Lipofectamine. An inducible cell line which can express antisense RNA of survivin with 2 mmol/L Zn(2 ) was obtained by the selection with 800 nmol/L G418 solution. The expression of antisense RNA of survivin inhibited the expression of survivin protein. Inhibition of survivin expression suppressed the proliferation of HeLa cells and its cell cycle. And it also sensitized HeLa cells to chemotherapeutics.
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To study the function of survivin in cancer cells, survivin cDNA was amplified from HeLa cells by RT-PCR. Then the coding fragment was subcloned into an inducible eukaryotic expression plasmid pHC in reverse direction.The obtained plasmid pHSC was transfected into HeLa cells by Lipofectamine. An inducible cell line which can express antisense RNA of survivin with 2 mmol/L Zn(2 ) was obtained by the selection with 800 nmol/L G418 solution. The expression of antisense RNA of survivin inhibited the expression of survivin protein. Inhibition of survivin expression suppressed the proliferation of HeLa cells and its cell cycle. And it also sensitized HeLa cells to chemotherapeutics.
Key concepts: Survivin, HeLa, Lipofectamine, Antisense RNA, Molecular biology, Transfection, RNA, Chemistry