斑蝥酸钠在诱导人宫颈癌Hela细胞凋亡相关基因表达中的作用机制研究

The mechanism of sodium cantharidinate in inducing apoptosis related gene expression in human cervical cancer Hela cells

目的:探讨斑蝥酸钠在诱导人宫颈癌Hela细胞凋亡相关基因表达中的作用机制。方法:2023年1月—2023年12月购买1株人宫颈癌Hela细胞,培养至对数生长期后开始实验,将其分为4组,分别加斑蝥酸钠0μmol/L(空白对照组)、4μmol/L(4μmol/L组)、8μmol/L(8μmol/L组)和16μmol/L(16μmol/L组),每个浓度分别设置5个复孔(每组样本量为5),以细胞活力试验(CCK-8)检测细胞增殖情况,以流式细胞仪检测细胞凋亡情况,以实时荧光定量多聚核苷酸链式反应(Real-time PCR)检测B淋巴细胞瘤2(BCL-2)、人细胞凋亡调节因子(Bax)、人半胱氨酸蛋白酶3(Caspase-3)的表达情况。结果:4μmol/L组、8μmol/L组和16μmol/L组的细胞增殖抑制率、凋亡率和Caspase-3相对表达量水平均高于空白对照组,BCL-2/Bax相对表达量水平低于空白对照组,差异均有统计学意义(P<0.05)。随着斑蝥酸钠浓度的升高,人宫颈癌Hela细胞增殖抑制率、凋亡率和Caspase-3随之也升高,BCL-2/Bax相对表达量逐渐下降,差异均有统计学意义(P<0.05)。经斑蝥酸钠处理后可见细胞皱缩、变圆,细胞边缘出芽形成多花环结构的凋亡小体。结论:斑蝥酸钠具有抑制人宫颈癌Hela细胞增殖、促进其凋亡的作用,具有剂量依赖性,其对细胞凋亡的调控作用可能与调控BCL-2,Bax和Caspase-3表达有关。

Objective:To investigate the mechanism of sodium cantharidinate in inducing apoptosis related gene expression in human cervical cancer Hela cells.Methods:Purchase one human cervical cancer Hela cell line from January 2023 to December 2023,culture it until the logarithmic growth phase,and start the experiment.Divide it into four groups,add sodium cantharidinate 0 and 4 separatelyμmol/L,8μmol/L,16μmol/L,set as blank control group and 4μmol/L group,8μmol/L group,16μmol/L group,5 wells were set for each concentration(with a sample size of 5 in each group).Cell proliferation was detected using cell viability assay(CCK-8),cell apoptosis was detected using flow cytometry,and the expression of B lymphoblastoma 2(BCL-2),human apoptosis regulatory factor(Bax),and human cysteine protease 3(Caspase-3)was detected using real-time fluorescence quantitative polymerase chain reaction(Real time PCR).Results:The cell proliferation inhibition rate,apoptosis rate and relative expression level of Caspase-3 in 4μmol/L Group,8μmol/L Group and 16μmol/L Group were higher than those in the blank control group,while the relative expression level of BCL-2/Bax was lower than that in the blank control group,the difference was significant(P<0.05).With the increase of the concentration of sodium cantharidin,the proliferation inhibition rate,apoptosis rate and Caspase-3 of human cervical carcinoma Hela cells also increased,and the relative expression of BCL-2/Bax gradually decreased(P<0.05).After treated with sodium cantharidin,the cells shrank,became round,and the edge of the cells sprouted to form many garland-like apoptotic bodies.Conclusion:Sodium cantharidinate has a dose-dependent effect on inhibiting the proliferation and promoting apoptosis of human cervical cancer Hela cells.Its regulatory effect on cell apoptosis may be related to the regulation of BCL-2,Bax,and Caspase-3 expression.