大蒜素联合热疗对人肺癌H1299细胞放疗增敏效果

Effect of allicin combined with hyperthermia on radiosensitization of human lung cancer H1299 cells

探讨了大蒜素联合热疗对人肺癌H1299细胞放疗敏感性的影响.体外培养人肺癌H1299细胞,实验分为五组:对照组、大蒜素处理组、热疗组、化疗组、大蒜素+热疗+化疗组,采用MTT法检测细胞存活率;克隆形成实验检测细胞增殖能力;流式细胞术检测细胞周期、细胞凋亡和线粒体膜电位(ΔΨm);qRT-PCR和Western blot检测PCNA,Bax,Bcl-2基因表达水平.结果显示,与对照组相比,大蒜素对H1299细胞生长具有剂量依赖性抑制作用,作用48 h的最佳质量浓度是20µg/mL.之后分别用20µg/mL大蒜素、45℃加热、8GyX线照射处理细胞,结果表明,三种处理均可降低细胞活力和克隆形成率;三者联合处理后细胞周期阻滞于G1期,细胞早期凋亡率为(4.75±0.28)%(P<0.01),晚期凋亡率为(19.30±0.49)%(P<0.01),ΔΨm下降至3.71±0.50(P<0.01),增殖标志基因PCNA表达降低,凋亡相关Bax基因表达升高,Bcl-2基因表达降低(P<0.01).大蒜素联合热疗共同作用人肺癌H1299细胞可增加其放疗敏感性,通过上调Bax,下调Bcl-2诱导其凋亡.

It was investigated the effect of allicin combined with hyperthermia on radiosensitization of human lung cancer H1299 cells.Human lung cancer H1299 cells were culturedin vitro and divided into five groups:control group,allicin treatment group,hyperthermia group,chemotherapy group and allicin+hyperthermia+chemotherapy group.The cell survival rate was detected by MTT assay.Clonal formation assay was used to detect cell proliferation.The cell cycle,apoptosis and mitochondrial membrane potential(ΔΨm)were detected by flow cytometry.The expression levels of PCNA,Bax and Bcl-2 genes were detected by qRT-PCR and Western blot.The results showed that compared with the control group,allicin had a dose-dependent inhibitory effect on H1299 cell growth,and the optimal quality concentration for 48 h was 20µg/mL.After treatment with 20µg/mL allicin,45℃heating and 8 Gy X-ray irradiation,cell viability and clone formation rate were decreased.The results showed that all three treatments could reduce cell viability and clone formation rate.The cell cycle was blocked in G1 phase,the apoptosis rate in early stage was(4.75±0.28)%(P<0.01),the apoptosis rate in late stage was(19.30±0.49)%(P<0.01),ΔΨm decreased to 3.71±0.50(P<0.01),and the expression of proliferation marker gene PCNA decreased.Apoptosis-related Bax gene expression increased,Bcl-2 gene expression decreased(P<0.01).Allicin combined with hyperthermia can increase the radiosensitization of human lung cancer H1299 cells and induce apoptosis by up-regulating Bax and down-regulating Bcl-2.