力达霉素对人宫颈癌Caski细胞增殖的影响及其机制

摘要

Objective To investigate the effect of lidamycin on the proliferation of human cervical cancer Caski cells and its underlying molecular mechanisms.Methods Human cervical cancer Caski cells were randomly divided the experimental groups (including the low-dose,medium-dose and high-dose groups) which were separately treated with 1,2.5 and 5 ng/mL lidamycin (100 μL) for 24 h,and the control group which was added with the same volume of RPMI-1640.MTT assay was used to detect the inhibition rate of proliferation.The cell cycle and mitochondrial membrane potential of CasKi cells were measured by flow cytometry.Transmission electronic microscope was used to observe the morphological changes of Caski cells.We calculated the apoptosis rate and mitotic catastrophe rate.Results The proliferation rate and survival rate of the experimental groups were significantly lower than those of the control group,and the decrease in the high-dose group was more significant (all P ＜ 0.05).The percentage of cells in the S phase of the control group and the medium-dose group was higher than those in the low-dose and high-dose groups (all P ＜ 0.05).The percentage of cells in the G0/G1 phase of the experimental groups was lower than that of the control group and the percentage of cells in the G2/M phase was higher than that of the control group,especially in the medium-dose and high-dose groups (all P ＜ 0.05).The ratios of mitochondrial membrane potential decrease in the low-dose,medium-dose and high-dose groups and control group were 34.77％ ± 5.33％,37.87％ ± 5.73％,18.37％ ± 3.91％,and 4.17％ ± 0.65％,respectively.The mitochondrial membrane potential decreased ratios of the experimental groups were higher than that in the control group,especially in the lowdose and medium-dose groups (all P ＜ 0.05).Transmission electronic microscopy showed that the control group had normal morphology while the experimental groups had many forms of death cells,including apoptosis,necrosis and mitotic catastrophe.The apoptosis rate and mitotic catastrophe rate of the experimental groups were both higher than those of control group (both P ＜ 0.05).Conclusion Lidamycin can significantly inhibit the proliferation of Caski cells with an dose-dependent manner by inducing the apoptosis and mitotic catastrophe of cells.%目的 探讨力达霉素对人宫颈癌Caski细胞增殖的影响及其可能的机制.方法 将对数生长期的人宫颈癌Caski细胞随机分为低、中、高浓度组及对照组,低、中、高浓度组均加入含力达霉素的RPMI 1640完全培养基100μL,分别调整其终浓度为1、2.5、5 ng/mL,对照组仅加入等体积RPMI 1640完全培养基.作用24h,采用MTT法检测细胞增殖能力(以吸光度值表示),计算细胞生存率;采用流式细胞术检测细胞周期及线粒体膜电位降低比例;透射电子显微镜下观察细胞形态学改变,计算细胞凋亡率及裂亡率.结果 低、中、高浓度组细胞增殖能力及细胞生存率均低于对照组,且高浓度组降低更明显(P均＜0.01).对照组及中浓度组S期细胞比例均高于低、高浓度组(P均＜0.05).低、中、高浓度组G0/G1期细胞比例均低于对照组,G2/M期细胞比例均高于对照组,且中、高浓度组变化更明显(P均＜0.05).低、中、高浓度组及对照组细胞线粒体膜电位降低比例分别为(34.77±5.33)％、(37.87±5.73)％、(18.37±3.91)％、(4.17±0.65)％;低、中、高浓度组细胞线粒体膜电位降低比例均高于对照组,且低、中浓度组均高于高浓度组(P均＜0.05).透射电镜下可见对照组细胞形态正常,低、中、高浓度组存在多种形态的死亡细胞,包括凋亡、坏死、裂亡的细胞.对照组及高、低、中浓度组细胞凋亡率及裂亡率均依次升高,两组间比较P均＜0.05.结论 力达霉素可抑制人宫颈癌Caski细胞增殖,并呈浓度依赖性,其机制可能与诱导细胞凋亡或裂亡有关.