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[目的]探讨核因子-κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸(PDTC)对胃癌细胞MKN-45 p53异构体表达的影响及其生物学意义。[方法 ]不同浓度的PDTC(25、50、75μmol/L)单独及联合顺铂(4μg/ml)作用于胃癌细胞MKN-45,采用细胞增殖/毒性检测试剂盒(CCK-8试剂盒)检测细胞增殖抑制率;实时荧光定量多聚酶链反应(RT-PCR)检测p53β、Δ133p53及NF-κB p65在m RNA水平的表达情况。[结果 ]CCK-8结果显示,PDTC(25、50、75μmol/L)单独作用时能抑制MKN-45细胞生长,抑制率分别为4.95%、12.20%、21.28%,差异有统计学意义(P<0.0001);当与顺铂(4μg/ml)联合时,MKN-45细胞增殖抑制率随PDTC浓度的增加而增加,且高于单独顺铂组,差异有统计学意义(P<0.001)。RT-PCR结果显示,当用不同浓度PDTC(25、50、75μmol/L)预处理2h后再加入顺铂(4μg/ml),MKN-45细胞中p53βm RNA的表达差异无统计学意义(P=0.04),而Δ133p53和p65 m RNA的表达随PDTC浓度的增加而降低,且低于顺铂单独组,差异有统计学意义(P<0.0001)。Pearson相关性分析显示,p65与p53βm RNA表达无相关性(r=0.072,P=0.798),p65与Δ133p53 m RNA表达呈正相关(r=0.814,P<0.0001)。[结论 ]NF-κB抑制剂PDTC可以抑制MKN-45细胞的生长,也可以增强顺铂对该细胞的生长抑制效应,Δ133p53异构体可能是NF-κB-p53对话的关键分子。
[Objective] To investigate the effect of pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor-κB (NF-κB), on p53 isoform p53 expression in gastric cancer cells and its biological significance. [Method] The gastric cancer cell line MKN-45 was treated with different concentrations of PDTC (25, 50 and 75μmol / L) alone and in combination with cisplatin (4μg / ml) and detected by the cell proliferation / toxicity assay kit (CCK-8 kit) Cell proliferation inhibition rate; the expression of p53β, Δ133p53 and NF-κB p65 at m RNA level were detected by real-time fluorescent quantitative polymerase chain reaction (RT-PCR). [Results] The results of CCK-8 showed that PDTC (25, 50, 75μmol / L) alone inhibited the growth of MKN-45 cells with the inhibition rates of 4.95%, 12.20% and 21.28%, respectively <0.0001). When combined with cisplatin (4μg / ml), the proliferation inhibition rate of MKN-45 cells increased with the increase of PDTC concentration, which was higher than that of cisplatin alone (P <0.001). The results of RT-PCR showed that the expression of p53βmRNA in MKN-45 cells was not significantly different after cisplatin (4μg / ml) pretreatment with different concentrations of PDTC (25, 50 and 75μmol / L) = 0.04). However, the expression of Δ133p53 and p65mRNA decreased with the increase of PDTC concentration and was lower than that of cisplatin alone (P <0.0001). Pearson correlation analysis showed that there was no correlation between p65 and p53βmRNA expression (r = 0.072, P = 0.798). There was a positive correlation between p65 and Δ133p53m RNA expression (r = 0.814, P <0.0001). [Conclusion] PDTC, an inhibitor of NF-κB, can inhibit the growth of MKN-45 cells and enhance the inhibitory effect of cisplatin on it. The Δ133p53 isoform may be the key molecule of NF-κB-p53.