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目的探讨胃癌细胞株是否存在核转录因子(NF)-κB组成性激活及对胃癌细胞增殖的影响。方法采用逆转录-聚合酶链反应(RT-PCR法)和蛋白免疫印迹法(Westernblot法)检测4株不同分化程度的胃癌细胞株NF-κB的mRNA和蛋白表达,比较4株胃癌细胞株中NF-κB的转录和蛋白质表达水平的差异,并利用TransAMTMNF-κBP65试剂盒比较不同胃癌细胞株中P65亚基的蛋白活性差异。采用噻唑蓝(MTT)比色法观察NF-κB活性抑制剂PDTC(吡咯烷二硫代氨基甲酸盐)对胃癌细胞增殖的影响。结果4株胃癌细胞株中均存在NF-κB的组成性激活,且存在蛋白表达和活性差异。活化的P65亚基在MKN28、MKN45细胞株中表达较低,在AGS、SGC-7901细胞株中表达较高。经PDTC处理的实验组胃癌细胞的增殖明显受到抑制,对照组呈正常生长。结论胃癌细胞株中存在NF-κB组成性激活及P65蛋白表达差异。抑制NF-κB的活性可明显抑制胃癌细胞的增殖。
Objective To investigate whether gastric cancer cell lines constitutively activate nuclear factor-kappa B (NF-κB) and affect the proliferation of gastric cancer cells. Methods The mRNA and protein expressions of NF-κB in 4 gastric cancer cell lines with different differentiation degrees were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting NF-κB transcription and protein expression level difference, and the use of TransAMTMNF-κBP65 kit compared different gastric cancer cell lines P65 subunit protein activity differences. The effect of NF-κB inhibitor PDTC (pyrrolidine dithiocarbamate) on the proliferation of gastric cancer cells was observed by MTT assay. Results The constitutive activation of NF-κB existed in all four gastric cancer cell lines, and there were differences in protein expression and activity. Activated P65 subunit was lower in MKN28 and MKN45 cell lines and higher in AGS and SGC-7901 cell lines. The proliferation of gastric cancer cells treated with PDTC was significantly inhibited, while the control group showed normal growth. Conclusion The constitutive activation of NF-κB and the difference of P65 protein expression exist in gastric cancer cell lines. Inhibition of NF-κB activity can significantly inhibit the proliferation of gastric cancer cells.