黄秋葵多糖调控p73反义RNA 1T对宫颈癌细胞增殖和凋亡的影响

目的探讨黄秋葵多糖调控p73反义RNA 1T(TP73-AS1)对宫颈癌细胞增殖和凋亡的影响,及其作用机制。方法实验分为对照组(正常DMEM培养基),低、中、高剂量实验组(分别用含0.6,1.2,1.8 mg·mL^-1黄秋葵多糖的DMEM培养基)、si-NC组(转染TP73-AS1阴性对照质粒)、si-TP73-AS1组(转染TP73-AS1抑制表达质粒)、pcDNA3.1组(1.2 mg·mL^-1黄秋葵多糖+TP73-AS1阳性对照质粒)、pcDNA3.1-TP73-AS1组(1.2 mg·mL^-1黄秋葵多糖+TP73-AS1过表达质粒)。用四甲基偶氮唑盐比色法检测细胞存活率,用流式细胞术检测细胞凋亡,用实时荧光定量聚合酶链反应检测TP73-AS1的表达水平。结果低、高剂量实验组和对照组、si-NC组、si-TP73-AS1组、pcDNA3.1组、pcDNA3.1-TP73-AS1组的细胞存活率分别为(71.67±3.08)%,(37.36±2.27)%,(100.55±3.68)%,(100.91±6.81)%,(57.12±4.98)%,(49.26±2.81)%和(81.19±7.09)%,细胞凋亡率分别为(14.67±1.40)%,(25.17±2.59)%,(8.38±1.22)%,(8.98±1.04)%,(19.42±1.90)%,(21.13±2.16)%和(10.23±1.06)%,TP73-AS1表达水平分别为0.67±0.07,0.28±0.03,1.00±0.10,1.01±0.10,0.52±0.05,0.42±0.04和0.76±0.06。低、高剂量实验组的上述指标与对照组相比,si-TP73-AS1组的上述指标与si-NC组相比,pcDNA3.1-TP73-AS1组的上述指标与pcDNA3.1组相比,差异均有统计学意义(均P<0.05)。结论黄秋葵多糖可抑制宫颈癌细胞增殖,促进细胞凋亡,其机制可能与下调TP73-AS1有关。

Effect of raw polysaccharide on the proliferation and apoptosis of cervical carcinoma cells by regulating p73 antisense RNA 1T

Objective To investigate the effects of raw polysaccharide on the proliferation and apoptosis of cervical cancer cell by regulating p73 antisense RNA 1 T(TP73-AS1).Methods The experiment was divided into control group(normal DMEM medium),experimental-L,-M,-H groups(0.6,1.2,1.8 mg·mL^-1raw polysaccharide),si-NC group(transfected with TP73-AS1 negative control plasmid),si-TP73-AS1 group(transfected with TP73-AS1 inhibition expression plasmid),pcDNA3.1 group(1.2 mg·mL^-1raw polysaccharide+TP73-AS1 positive control plasmid),and pcDNA3.1-TP73-AS1 group(1.2 mg·mL^-1raw polysaccharide+TP73-AS1 overexpression plasmid).The tetramethylazozolium colorimetry was used to detect the cell survival rate.The flow cytometry was used to detect the cell apoptosis.The expression of TP73-AS1 was detected by real-time fluorescence quantitative polymerase chain reaction.Results The cell survival rates of experimental-L,experimental-H,control,si-NC,si-TP73-AS1,pcDNA3.1,pcDNA3.1-TP73-AS1 groups were(71.67±3.08)%,(37.36±2.27)%,(100.55±3.68)%,(100.91±6.81)%,(57.12±4.98)%,(49.26±2.81)%and(81.19±7.09)%,apoptosis rates were(14.67±1.40)%,(25.17±2.59)%,(8.38±1.22)%,(8.98±1.04)%,(19.42±1.90)%,(21.13±2.16)%and(10.23±1.06)%,the expression levels of TP73-AS1 were 0.67±0.07,0.28±0.03,1.00±0.10,1.01±0.10,0.52±0.05,0.42±0.04 and0.76±0.06.There were statistically significant differences between experimental-L,-H groups and control group,si-TP73-AS1 group and si-NC group,pcDNA3.1-TP73-AS1 group and the pcDNA3.1 group(all P<0.05).Conclusion Raw polysaccharide can inhibit the proliferation of cervical cancer cells and promote apoptosis,which may be related to the down-regulation of TP73-AS1 expression.