摘要:
Objective To investigate the effects of Hydroxycamptothecin (HCPT) on the cervical cancer cells transfected with wild-type p53. Methods HeLa cells were cultured in vitro, and some were transfected with the plasmid pcDNA3.1-HA-EGFP (negative control group) or the target plasmid pcDNA3.1-HA-TP53 (experimental group). Then the cells were divided into 5 groups: blank control group, sol-vent group, HCPT group, p53 group and HCPT+p53 group. The expression of the p53 gene was detected by qRT-PCR, CCK-8 assay was used to detect the viability of cells treated with different concentrations of HCPT. The apoptosis rates of different groups were measured by flow cytometry. Results Under the fluorescent microscope, there was obvious green fluorescence in the cells, indicating that the plasmid was successfully transfected. After the HeLa cells were transfected with the target plasmid, the mRNA expression of p53 gene was up-regulated along with the increase of the transfection plasmid dose. Compared with the blank and negative control groups, the viability of cells in exper-imental groups was inhibited in a HCPT-dose dependent manner, and there were statistical differences when it treated with different doses of HCPT (P<0.05). Compared with the blank control group and solvent group, the apoptosis rates of HCPT group and p53 group were increased (P<0.05), and that of the p53+HCPT group was significantly higher than that of the other experimental groups (P<0.05). Conclusion The introduction of exogenous wild-type p53 gene into cervical cancer HeLa cells can significantly enhance the effect of HCPT in inhibiting the proliferation of cervical cancer cells and promoting its apoptosis.%目的 探讨羟喜树碱(Hydroxycamptothecin,HCPT)对外源野生型p53转染宫颈癌细胞的作用效果.方法 体外培养宫颈癌HeLa细胞,将其分为空白对照组、阴性对照组、HCPT组、p53组、HCPT+p53组.荧光显微镜下观察转染结果,qRT-PCR检测p53基因的表达,CCK-8法检测不同浓度HCPT作用于目的质粒转染后细胞的活力,流式细胞技术检测细胞凋亡率.结果 荧光显微镜下可见细胞内有明显绿色荧光,说明质粒转染成功;HeLa细胞转染目的质粒后,p53基因mRNA表达上调,且随着转染质粒剂量的增加而逐渐升高;与空白组、阴性对照组相比,实验组细胞存活率降低,随着HCPT浓度的增加,对细胞活力的抑制作用明显增加,各浓度比较,差异有统计学意义(P<0.05);与空白组和溶剂组相比,HCPT组、p53组的凋亡率升高,差异具有统计学意义(P<0.05);p53+HCPT组凋亡率显著高于其他实验组,差异具有统计学意义(P<0.05).结论 外源野生型p53基因导入宫颈癌HeLa细胞后,可明显增强HCPT的作用,抑制HeLa细胞的增殖,促进其凋亡.HCPT和p53可协同抑制HeLa细胞的增殖并促进其凋亡.