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Jiang Z.,Weifang Institute of Traditional Chinese Medicine | Jiang Z.,Weifang Medical University | Sun C.,Weifang Institute of Traditional Chinese Medicine | Zhuang J.,Weifang Institute of Traditional Chinese Medicine | Liu R.,Weifang Institute of Traditional Chinese Medicine
Chinese Journal of Clinical Oncology | Year: 2013

Objective: To investigate if PUMA gene transfection could increase the sensitivity of breast cancer cells (MCF-7) on Epirubicin-induced apoptosis. Methods: PUMA-pCDNA3, which contains full length PUMA cDNA or -pCDNA3, was transfected into the human breast cancer cell line MCF-7 by using lipofectamine. G418 selection was used to select positive cells. MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells were respectively treated with serial concentrations of Epirubicin (0.01 μmol/L to 100 μmol/L). MTT assay was performed to determine the cell survival rate in each group. IC50 of Epirubicin was then calculated. TUNEL and FCM were employed to investigate cell apoptosis. Western blot was performed to detect PUMA protein expression. Results: The Epirubicin IC50 values of MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells were (13±1.4), (1.8±0.2), and (10.7±1.3) μmol/L, respectively. The sensitivity of MCF-7/PUMA cells had a 7.2-fold increase. The Epirubicin-induced apoptosis of MCF-7 cells behaved in a dose-dependent manner. The apoptosis of MCF-7/ PUMA cells was more prominent than that of MCF-7 and MCF-7/ pCDNA3 cells. At a low concentration (0.1 μmol/L), Epirubicin could only slightly induce MCF-7/ pCDNA3 cells [(1.15±0.26)%] and MCF-7 cells [(0.9±0.24)%], as well as the apoptosis in MCF-7/PUMA cells [(6.44±1.46)%J. At a high concentration, Epirubicin (1.0 μmol/L) could induce apoptosis in all groups. The apoptotic rate was significantly higher in MCF-7/PUMA cells [(35.47±9.36)%] than in MCF-7/pCDNA3 cells [(15.2±5.17)%] and in MCF-7 cells [(12.6±3.73)%], both P<0.01). Results obtained from FCM and TUNEL were the same. PUMA protein expression was significantly higher in MCF-7/PUMA cells than in MCF-7 and MCF-7/pCDNA3 cells. Conclusion: PUMA gene transfection greatly enhances the sensitivity of MCF-7 cells during Epirubicin-induced apoptosis. Source


Zhuang J.,Nanjing University of Traditional Chinese Medicine | Sun C.,Weifang Institute of Traditional Chinese Medicine | Wang C.,Weifang Medical University | Feng F.,Weifang Medical University | And 3 more authors.
Chinese Journal of Clinical Oncology | Year: 2014

Objective: This study aimed to explore the effects of trichosanthin on the proliferative inhibition and apoptosis induction in human lung carcinoma A549 cells. Methods: A549 cells were treated with various concentrations of TCS. The inhibitory effects in proliferation were detected by the MTT method. The microfilament changes were observed by transmission electron microscopy. Apoptosis rate and cell cycle were determined by flow cytometry. Results: A549 cells treated with TCS presented apoptotic changes and decreased cell activity. When the concentration increased and time was prolonged, the inhibition rate increased correspondingly. Conclusion: Pharmacological doses of TCS inhibited the proliferation and differentiation in lung carcinoma A549 cells and affected the function in A549 cells by changes in the cytoskeleton. Source

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