目的研究端粒酶反义寡核苷酸在肺腺癌A549细胞中抑制端粒酶活性和诱导细胞凋亡的能力。方法用脂质体将荧光标记的端粒酶反义寡核苷酸转染入肺腺癌A549细胞后,TR AP-PC R-ELISA检测端粒酶活性变化,凋亡细胞用透射电镜(TEM)观察,TU N EL...目的研究端粒酶反义寡核苷酸在肺腺癌A549细胞中抑制端粒酶活性和诱导细胞凋亡的能力。方法用脂质体将荧光标记的端粒酶反义寡核苷酸转染入肺腺癌A549细胞后,TR AP-PC R-ELISA检测端粒酶活性变化,凋亡细胞用透射电镜(TEM)观察,TU N EL法检测各组细胞凋亡率。结果转染72h后,端粒酶反义寡核苷酸进入A549细胞内,端粒酶活性显著降低。透射电镜下,端粒酶反义寡核苷酸转染组细胞凋亡增加,细胞凋亡率比其他组明显增高,差异有统计学意义(P<0.01)。结论端粒酶反义寡核苷酸能靶向端粒酶,抑制其活性,并诱导肺腺癌A549细胞凋亡。展开更多
To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the w...To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the wild PTEN gene expression plasmid (pGFP-PTEN) by magnetic iron nanoparticle and lipo2000. The transfection efficiency was detected by fluorescence microscope and flow cytometer. The expression levels of PTEN mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry analysis. The effect of PTEN transfection on cell cycle enhances the sensitivity of A549/CDDP to cisplatin and nanoparticle-mediated transfection has a higher efficiency than that of the liposome-mediated group. The apoptosis level was up-regulated in PTEN transfection group. The magnetic iron oxide nanoparticle could be used as one of the ideal gene carriers for PTEN gene delivery in vitro. PTEN can be an effective target for reversing cisplatin-resistance in lung cancer.展开更多
基金Natural Science Foundation of Hunan,China(No.07JJ3055)
文摘目的研究端粒酶反义寡核苷酸在肺腺癌A549细胞中抑制端粒酶活性和诱导细胞凋亡的能力。方法用脂质体将荧光标记的端粒酶反义寡核苷酸转染入肺腺癌A549细胞后,TR AP-PC R-ELISA检测端粒酶活性变化,凋亡细胞用透射电镜(TEM)观察,TU N EL法检测各组细胞凋亡率。结果转染72h后,端粒酶反义寡核苷酸进入A549细胞内,端粒酶活性显著降低。透射电镜下,端粒酶反义寡核苷酸转染组细胞凋亡增加,细胞凋亡率比其他组明显增高,差异有统计学意义(P<0.01)。结论端粒酶反义寡核苷酸能靶向端粒酶,抑制其活性,并诱导肺腺癌A549细胞凋亡。
基金Project(07JJ3055)supported by the Natural Science Foundation of Hunan Province,China
文摘To evaluate the feasibility of using magnetic iron oxide nanoparticle as wild PTEN gene carrier for transfection in vitro to reverse cisplatin-resistance of A549/CDDP cells, A549/CDDP cells were transfected with the wild PTEN gene expression plasmid (pGFP-PTEN) by magnetic iron nanoparticle and lipo2000. The transfection efficiency was detected by fluorescence microscope and flow cytometer. The expression levels of PTEN mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry analysis. The effect of PTEN transfection on cell cycle enhances the sensitivity of A549/CDDP to cisplatin and nanoparticle-mediated transfection has a higher efficiency than that of the liposome-mediated group. The apoptosis level was up-regulated in PTEN transfection group. The magnetic iron oxide nanoparticle could be used as one of the ideal gene carriers for PTEN gene delivery in vitro. PTEN can be an effective target for reversing cisplatin-resistance in lung cancer.
