Effect of shRNA-mediated survivin gene silencing on sensitivity of lymphoma cells to adriamycin |
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Authors: | GU Cong-min ZENG Mu LIN Han-liang |
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Institution: | 1.Department of Pathology, Guangzhou Women and Children Hospital, Guangzhou 510180, China;2.Continuous Education College, Jinan University, Guangzhou 510630, China;3.Department of Pathology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China. E-mail:gucongmin@yahoo.com.cn |
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Abstract: | AIM: This study was designed to use RNA interference technique to down-regulate the expression of survivin gene in human Burkitts lymphoma cell line Daudi and to explore the effect on sensitivity of Daudi cells to adriamycin. METHODS: The survivin-shRNA expression vector was constructed and transfected into Daudi cells. Expression of survivin mRNA and protein were assessed by RT-PCR and Western blotting analysis, respectively. Apoptosis index of transfected Daudi cells was quantified by flow cytometry. The sensitivity of Daudi cells to adriamycin (ADR) before and after transfection was detected by MTT test. RESULTS: The mRNA and protein levels of survivin were down-regulated by 62.32% and 61.88%, respectively, compared to those in control-shRNA treated group and PBS treated group (P<0.05). Meanwhile, the apoptosis index was significantly increased (19.10%±2.15%), compared to that in control group (4.48%±1.54%) and PBS group (4.35%±1.37%, P<0.05). The 50% inhibition concentration (IC50) of ADM to Daudi cells was significantly decreased (0.25±0.43) μmol/L, compared to that in control group (0.87±0.21) μmol/L and PBS group (0.91±0.36) μmol/L, P<0.05. CONCLUSION: Down-regulation of survivin expression in Daudi cells by shRNA effectively induces apoptosis and increases the sensitivity of Daudi cells to ADR. |
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Keywords: | RNA interference Short-hairpin RNA Survivin Daudi cells |
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