Antisense RNA of survivin enhances the sensitivity of pancreatic cancer cell line PANC-1 to doxorubicin
Jinghua Shen, Xiaojuan Wang, Xiumei Wang, He-ba-te Su
Abstract
Jinghua Shen, Xiaojuan Wang, Xiumei Wang, He-ba-te Su
Abstract
This paper attempts to discuss the effects of surviving antisense RNA on doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-1. A surviving antisense eukaryotic vector pcDNA3-SV Vas prepared in previous study was delivered into PANC-1 by electroperforation. Cell survival fraction and MTT assay were used to investigate the sensibility of transfected cells to doxorubicin. Apoptosis was detected by DNA gel electrophoresis. We obtained two positive cell clone PANC—1/SVVas and PANC-1/neo cells, the growth of PANC-1/SV Vas cells was significantly reduced ( P <0.05). By MTT assay, the IC 50 to doxorubicin of PANC-1/SV Vas, PANC-1/neo and PANC-1 cells were (0.285±0.012) μ mol/L, (1.528±0.317) μ mol/L and (1.540±0.253) μ mol/L respectively, the difference was significant by statistic analysis ( P <0.01). Agarose gel electrophoresis of genomic DNA from PANC/SV Vas showed typical DNA ladder, but DNA from PANC-1/neo and PANC-1 did not. Survivin antisense RNA could enhance doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-1. This may lay an experimental foundation for further research of gene therapy in pancreatic cancer.
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This paper attempts to discuss the effects of surviving antisense RNA on doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-1. A surviving antisense eukaryotic vector pcDNA3-SV Vas prepared in previous study was delivered into PANC-1 by electroperforation. Cell survival fraction and MTT assay were used to investigate the sensibility of transfected cells to doxorubicin. Apoptosis was detected by DNA gel electrophoresis. We obtained two positive cell clone PANC—1/SVVas and PANC-1/neo cells, the growth of PANC-1/SV Vas cells was significantly reduced ( P <0.05). By MTT assay, the IC 50 to doxorubicin of PANC-1/SV Vas, PANC-1/neo and PANC-1 cells were (0.285±0.012) μ mol/L, (1.528±0.317) μ mol/L and (1.540±0.253) μ mol/L respectively, the difference was significant by statistic analysis ( P <0.01). Agarose gel electrophoresis of genomic DNA from PANC/SV Vas showed typical DNA ladder, but DNA from PANC-1/neo and PANC-1 did not. Survivin antisense RNA could enhance doxorubicin-induced apoptosis in pancreatic cancer cell line PANC-1. This may lay an experimental foundation for further research of gene therapy in pancreatic cancer.
Key concepts: Survivin, Molecular biology, Doxorubicin, Apoptosis, Transfection, Agarose gel electrophoresis, MTT assay, Cell culture