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目的检测抗凋亡因子 XIAP 与 PED/PEA-15在前列腺癌细胞(PC-3)中的表达,探讨二者对前列腺癌细胞凋亡的影响。方法应用半定量 RT-PCR 法检测前列腺癌细胞(PC-3)中PED/PEA-15和 XIAP 的表达。设计并构建 PED/PEA-15和 XIAP 特异的 siRNA 载体,以脂质体法转染二者的 siRNA 载体至前列腺癌细胞(PC-3)中,半定量 RT-PCR 法检测特异 siRNA 载体对PED/PEA-15和 XIAP 转录的影响:光镜观察细胞形态改变;流式细胞法检测细胞凋亡的变化。结果半定量 RT-PCR 显示 PED/PEA-15和 XIAP 均在前列腺癌细胞(PC-3)中高表达。酶切和 DNA测序证实 XIAP 和 PED/PEA-15 siRNA 载体构建成功。共转染 XIAP 和 PED/PEA-15 siRNA 载体人 PC-3细胞,可导致 XIAP 和 PED/PEA-15的转录抑制,并增加 PC-3细胞对阿霉素的敏感性,凋亡明显增加,处理组凋亡率为79%,对照组为46%,两组差异有统计学意义(P<0.05)。结论 PED/PEA-15和 XIAP 在前列腺癌的凋亡中可起重要作用。
Objective To detect the expression of anti-apoptotic factor XIAP and PED / PEA-15 in prostate cancer cells (PC-3), and to explore their effects on the apoptosis of prostate cancer cells. Methods The expression of PED / PEA-15 and XIAP in prostate cancer cells (PC-3) was detected by semi-quantitative RT-PCR. PED / PEA-15 and XIAP specific siRNA vectors were designed and constructed. The siRNA vectors were transfected into prostate cancer cells (PC-3) by lipofectamine. The effect of specific siRNA vectors on PED / PEA-15 and XIAP transcription: The morphological changes of cells were observed under light microscope; the changes of apoptosis were detected by flow cytometry. Results Semi-quantitative RT-PCR showed that both PED / PEA-15 and XIAP were highly expressed in prostate cancer cells (PC-3). Digestion and DNA sequencing confirmed the successful construction of XIAP and PED / PEA-15 siRNA vectors. The co-transfection of XIAP and PED / PEA-15 siRNA vector human PC-3 cells can lead to the transcription inhibition of XIAP and PED / PEA-15 and increase the sensitivity of adriamycin-induced PC-3 cells to apoptosis, The apoptosis rate of the treatment group was 79% and that of the control group was 46%, the difference between the two groups was statistically significant (P <0.05). Conclusion PED / PEA-15 and XIAP may play an important role in the apoptosis of prostate cancer.