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目的:通过XIAP反义寡核苷酸转染hep-2细胞株,观察细胞增殖、凋亡及放疗反应。方法:体外培养人喉鳞状细胞癌hep-2细胞株,脂质体介导反义寡核苷酸转染,6h后4Gyγ射线照射。24h后RT-PCR检测XIAPmRNA表达,流式细胞仪检测蛋白水平及凋亡率,MTT检测细胞存活情况。结果:各浓度的反义寡核苷酸转染组hep-2细胞形态改变,MTT结果为细胞凋亡率随剂量增加而增高。800nmol/LXIAP反义寡核苷酸转染,使XIAPmRNA表达下调,蛋白表达下降,细胞凋亡率增加(P<0.05);4Gy照射后反义寡核苷酸转染组蛋白表达下降明显,较对照组细胞凋亡率增加,细胞存活率明显下降(P<0.05)。结论:XIAP反义寡核苷酸转染hep-2细胞株能够降低蛋白表达,促进细胞的凋亡并能提高对放疗的敏感性。
OBJECTIVE: To investigate the proliferation, apoptosis and radiotherapy of hep-2 cells by transfection with XIAP antisense oligonucleotides. Methods: Human laryngeal squamous cell carcinoma hep-2 cell line was cultured in vitro. Lipofectamine-mediated antisense oligonucleotide transfection was performed. 4Gy γ-ray irradiation was performed after 6 hours. The expression of XIAP mRNA was detected by RT-PCR 24h, the protein level and apoptosis rate were detected by flow cytometry, and the survival of cells was detected by MTT assay. Results: The morphological changes of hep-2 cells were observed in all concentrations of antisense oligonucleotide transfection group. MTT results showed that the apoptosis rate of hep-2 cells increased with the increase of dose. After transfection with 800 nmol / L XIAP antisense oligonucleotide, the expression of XIAP mRNA was down-regulated, the protein expression was decreased and the apoptosis rate was increased (P <0.05). The protein expression of antisense oligonucleotide transfected group decreased significantly after 4Gy irradiation, The apoptosis rate of control group increased, cell survival rate decreased significantly (P <0.05). CONCLUSION: Transfection of hep-2 cells with XIAP antisense oligonucleotides can reduce the protein expression, promote cell apoptosis and improve the sensitivity to radiotherapy.