苦参碱对宫颈癌SiHa细胞中自噬基因Beclin1和LC3Ⅱ表达的影响

Effect of matrine on the expression of Beclin1 and LC3Ⅱautophagy genes in cervical cancer SiHa cells

目的探究苦参碱对宫颈癌SiHa细胞的抑制作用以及对自噬基因Beclin1和LC3Ⅱ表达的影响。方法用含0(对照组),0.25(仅用于增殖抑制实验),0.50,1.00,2.00和4.00 mg·mL^-1苦参碱的RPMI-1640培养基体外培养宫颈癌SiHa细胞。干预24,48和72 h后,用噻唑蓝法检测细胞的增殖抑制;干预48 h后,用逆转录聚合酶链反应分别检测细胞中Beclin1和LC3ⅡmRNA的表达情况。结果苦参碱能够抑制宫颈癌SiHa细胞的增殖,且呈时间-剂量依赖性。干预48 h后,0.50,1.00,2.00,4.00 mg·mL^-1苦参碱组和对照组的抑制率分别为(19.83±2.06)%,(21.31±1.97)%,(48.13±2.96)%,(85.40±1.05)%和(1.02±0.02)%,Beclin1 mRNA分别为(8.41±0.05),(28.63±0.09),(36.79±0.06),(49.22±0.09)和1,LC3ⅡmRNA分别为(2.03±0.04),(3.16±0.03),(4.40±0.02),(5.23±0.06)和1,0.50,1.00,2.00,4.00 mg·mL^-1苦参碱组的上述指标与对照组比较,差异均有统计学意义(均P<0.05)。结论苦参碱可通过增加自噬基因Beclin1和LC3Ⅱ的表达,从而抑制宫颈癌SiHa细胞的增殖。

Objective To investigate the inhibitory effect of matrine on cervical cancer SiHa cells and its effects on theexpression of autophagy genes Beclin1 and LC3Ⅱ.Methods Cervical cancer SiHa cells were cultured in vitro on RPMI-1640 medium containing 0(control group),0.25(just for proliferation inhibition assay only),0.50,1.00,2.00 and 4.00 mg·mL^-1 matrine.After intervention 24,48 and 72 h,cell proliferation inhibition was determined using the method of methyl thiazolyl tetrazolium.After intervented 48 h,reverse transcription-polymerase chain reaction was used to detect the mRNA expression of Beclin1 and LC3Ⅱ.Results Matrine inhibited the proliferation of cervical cancer SiHa cells in a time-dose dependent manner.After intervention 48 h,the inhibition rates of 0.50,1.00,2.00,4.00 mg·mL^-1 matrine groups and control group were(19.83±2.06)%,(21.31±1.97)%,(48.13±2.96)%,(85.40±1.05)%and(1.02±0.02)%,the expressions of Beclin1 mRNA were(8.41±0.05),(28.63±0.09),(36.79±0.06),(49.22±0.09)and 1,the expressions of LC3ⅡmRNA were(2.03±0.04)(3.16±0.03),(4.40±0.02),(5.23±0.06)and 1.There were significant differences in the above-mentioned indexes between 0.50,1.00,2.00,4.00 mg·mL^-1 matrine groups and control group(all P<0.05).Conclusion Matrine can inhibit the proliferation of cervical cancer SiHa cells by increasing the expression of autophagy genes Beclin1 and LC3Ⅱ.