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目的:探讨抗氧化剂吡咯烷二硫代氨基甲酸盐(pyrrolidine dithiocarbamate,PDTC)联合卡铂对宫颈癌HeLa细胞增殖及凋亡的影响。方法:体外培养HeLa细胞,药物作用分为空白组、PDTC组、卡铂组和PDTC+卡铂联合组。各组药物处理后,MTT法检测细胞生长抑制率,FCM法检测细胞凋亡率和细胞周期分布情况,免疫细胞化学方法观察核转录因子κB(nuclearfactor-kappa B,NF-κB)p65在细胞质和细胞核中表达水平的变化。结果:MTT法检测结果显示,PDTC能够有效地抑制宫颈癌HeLa细胞的增殖,呈时间和剂量依赖关系;小剂量(12.5μmol/L)PDTC和卡铂联合应用较单用卡铂,能明显提高细胞生长抑制率(P<0.01)。FCM法检测结果显示,各用药组较对照组以及PDTC+卡铂联用组与单用卡铂组比较,宫颈癌HeLa细胞的凋亡率差异有统计学意义(P<0.01)。HeLa细胞经PDTC作用后,G0/G1期细胞比例较对照组明显增加(P<0.01),S期细胞比例降低(P<0.01);卡铂单用组使细胞周期阻滞于G2/M期;联合用药组使细胞周期进一步阻滞于G2/M期,且与对照组比较,降低了S期细胞比例(P<0.01)。免疫细胞化学法检测结果显示,NF-κBp65在宫颈癌HeLa细胞中主要表达于细胞质,卡铂诱导24 h后细胞质中NF-κBp65转移至细胞核,其活性增强,而PDTC能够抑制此作用。结论:卡铂能够诱导NF-κBp65的活化。小剂量(12.5μmol/L)PDTC和卡铂联合应用较单用卡铂,能明显增加对宫颈癌HeLa细胞的生长抑制率和细胞凋亡率。PDTC可能提高宫颈癌HeLa细胞对卡铂的敏感性。
Objective: To investigate the effect of pyrrolidine dithiocarbamate (PDTC) combined with carboplatin on the proliferation and apoptosis of cervical cancer HeLa cells. Methods: HeLa cells were cultured in vitro. The drug effects were divided into blank group, PDTC group, carboplatin group and PDTC + carboplatin group. The cell growth inhibition rate was detected by MTT assay and the apoptosis rate and cell cycle distribution were detected by FCM. The expression of nuclear factor-kappa B (NF-κB) p65 in cytoplasm and Changes in the expression level in the nucleus. Results: MTT assay showed that PDTC could effectively inhibit the proliferation of cervical cancer HeLa cells in a time-and dose-dependent manner. Combination of low dose (12.5 μmol / L) PDTC with carboplatin significantly increased Cell growth inhibition rate (P <0.01). The results of FCM assay showed that the apoptotic rates of cervical cancer HeLa cells were significantly different (P <0.01) compared with control group, PDTC + carboplatin group and carboplatin alone group. After HepG2 cells were treated with PDTC, the proportion of cells in G0 / G1 phase increased significantly (P <0.01) and the proportion of cells in S phase decreased (P <0.01); Cell cycle arrest in G2 / M phase ; The combination group further blocked the cell cycle G2 / M phase, and compared with the control group, reducing the proportion of S phase cells (P <0.01). The results of immunocytochemistry showed that NF-κBp65 mainly expressed in cytoplasm of cervical cancer HeLa cells, and the activity of NF-κBp65 was enhanced in cytoplasm 24 hours after the induction of carboplatin, while PDTC could inhibit this effect. Conclusion: Carboplatin can induce the activation of NF-κBp65. Low dose (12.5μmol / L) combination of PDTC and carboplatin compared with carboplatin alone significantly increased the growth inhibition rate and apoptosis rate of cervical cancer HeLa cells. PDTC may increase the sensitivity of cervical cancer HeLa cells to carboplatin.