论文部分内容阅读
目的 研究野生型p53基因对人胶质瘤细胞生长及化疗敏感性的影响。方法 将载有野生型p53基因的表达质粒PC53-SN3导入U251细胞。通过逆转录-聚合酶链反应(RT-PCR)检测p53基因在转染细胞中的表达,用四甲基偶氮唑盐微量酶反应比色法(MTT法)和流式细胞仪检测p53基因对U251细胞生长抑制及凋亡的影响以及它与顺铂联合作用后的效果。结果 通过RT-PCR证实了野生型p53基因在U251细胞中的表达。MTT检测发现p53基因对U251细胞的生长有明显的抑制作用,抑制率为(79.60±5.69)％。顺铂对 U251细胞的抑制作用随着顺铂浓度的增加(1、2、4、8 mg/L)而增强,抑制率分别为(19.40±6.69)％、(24.41±2.68)％、(51.84±13.38)％、(66.22±5.02)％,但不如 p53基因的抑制作用强。当野生型 p53基因与顺铂联合作用于 U251细胞时,抑制率明显增加,分别为(91.64±1.00)％、(94.98±1.67)％、(95.32±2.01)％、(95.65±1.00)％。p53基因转染所产生的 U251细胞凋亡率为 17.38％。顺铂引起的细胞凋亡率随着顺铂浓度的增加(1、2、4、8mg/L)而增加,分别为5.71％、5.93％、6.27％、6.81％。当p53基因与不同浓度的顺铂(1、2、4、8mg/L)联合作用于U251细胞时,凋亡率也明显增加,分别为23.50％、23.54％、23.89％、28.88％。结论 野生型p53基因与
Objective To investigate the effect of wild type p53 gene on the growth and chemosensitivity of human glioma cells. Methods The PC53-SN3 expression plasmid carrying wild-type p53 gene was introduced into U251 cells. The expression of p53 gene in transfected cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of p53 gene was detected by MTT assay and flow cytometry On U251 cell growth inhibition and apoptosis as well as its combined effect with cisplatin. Results The expression of wild-type p53 gene in U251 cells was confirmed by RT-PCR. The results of MTT assay showed that the p53 gene had a significant inhibitory effect on the growth of U251 cells with the inhibition rate of (79.60 ± 5.69)%. The inhibitory effect of cisplatin on U251 cells was enhanced with the increase of cisplatin concentrations (1, 2, 4 and 8 mg / L), and the inhibitory rates were (19.40 ± 6.69)%, (24.41 ± 2.68)% and ± 13.38)%, (66.22 ± 5.02)%, but not as strong as the p53 gene. When the wild type p53 gene combined with cisplatin on U251 cells, the inhibition rates were significantly increased (91.64 ± 1.00)%, (94.98 ± 1.67)%, (95.32 ± 2.01)% and (95.65 ± 1.00)%, respectively. The apoptosis rate of U251 cells generated by p53 gene transfection was 17.38%. Cisplatin-induced apoptosis increased with cisplatin concentrations (1, 2, 4 and 8 mg / L), which were 5.71%, 5.93%, 6.27% and 6.81%, respectively. When p53 gene was combined with different concentrations of cisplatin (1,2,4,8 mg / L) in U251 cells, the apoptosis rate was also significantly increased, respectively 23.50%, 23.54%, 23.89%, 28.88%. Conclusion Wild-type p53 gene and