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目的:通过研究纳秒脉冲消融与顺铂对人宫颈癌细胞Hela的体外作用,探讨低剂量纳秒脉冲电场联合低浓度顺铂之间的相互作用及机制。方法:采用CCK8法检测纳秒脉冲消融、顺铂对细胞增殖的影响,选取低于半抑制率(IC_(50))的低剂量(6个、12个)脉冲电场和低浓度(1μg/ml、2μg/ml)顺铂,CCK8法检测两者联合后的抑制率,计算联合作用指数(IC值),取IC值最低时的脉冲次数和顺铂浓度,将实验分为对照组、单纯脉冲电场组、单纯顺铂组和脉冲电场联合顺铂组,Annexin V/PI双染细胞后流式细胞仪检测凋亡率,Western blot法检测凋亡相关蛋白Bax、Bcl-2表达。结果:纳秒脉冲消融与顺铂均抑制细胞生长,低剂量脉冲电场和低浓度顺铂的联合作用指数均<0.9,表明两者有协同作用,脉冲电场联合顺铂组的凋亡率最高(P<0.05),且该组Bax蛋白表达最高,Bcl-2蛋白表达最低(P<0.05)。结论:低剂量脉冲电场联合低浓度顺铂具有协同作用,其中12个脉冲联合2μg/ml顺铂对肿瘤细胞抑制最显著,其机制可能与Bax高表达,促进细胞凋亡有关。
OBJECTIVE: To investigate the interaction between nanosecond pulse ablation and cisplatin on human cervical cancer cell Hela in vitro and to explore the interaction between low dose nanosecond pulse electric field and low concentration cisplatin and its mechanism. Methods: CCK8 method was used to detect the effects of ablation of nanosecond pulses and cisplatin on cell proliferation. Low dose (6, 12) pulsed electric fields with low half inhibitory rate (IC 50) , 2μg / ml) cisplatin and CCK8 assay, the combined effect index (IC) was calculated, and the pulse number and cisplatin concentration at the lowest IC value were used to divide the experiment into two groups: control group, Apoptosis rate was detected by flow cytometry after electric field, cisplatin alone, pulsed electric field combined with cisplatin and Annexin V / PI double staining. Western blot was used to detect the expression of Bax and Bcl-2. Results: The combination of nanosecond pulse ablation and cisplatin both inhibited the cell growth. The combined action index of low-dose pulsed electric field and low-concentration cisplatin were both <0.9, indicating that both had synergistic effect, and the apoptosis rate was the highest in the combination of pulsed electric field and cisplatin P <0.05), and the highest expression of Bax protein and lowest expression of Bcl-2 protein (P <0.05). CONCLUSION: Low-dose pulsed electric field combined with low-concentration cisplatin has a synergistic effect. Among them, 12 pulses combined with 2μg / ml cisplatin have the most significant inhibitory effect on tumor cells, which may be related to the high expression of Bax and cell apoptosis.