论文部分内容阅读
目的在rmh TNF-α-MKN45人胃癌细胞系模型中,观察NF-κB p65基因及蛋白表达的变化情况。方法以不同浓度的rmh TNF-α(0IU/ml,50IU/ml,100IU/ml,200IU/ml)作用人胃癌细胞系MKN45后,采用CCK-8法测定细胞增殖抑制率,巢式逆转录多聚酶链反应(RT-PCR)检测NF-κB p65基因的表达变化水平,并进行统计学分析,组间差异用单因素方差分析,组内比较用T-test。免疫荧光细胞化学(IF)方法检测NF-κB p65蛋白在胃癌细胞系MKN45的核转位变化情况。结果 CCK-8法结果显示,细胞增殖抑制率随rmh TNF-α浓度的增加而增加,差异有统计学差异;RT-PCR结果显示,随着rmh TNF-α浓度的增加,NF-κB p65 mRNA表达量下降,且组间差异具有统计学意义;IF结果显示,随着rmh TN浓度的增加NF-κB p65蛋白的核内活性下降。结论在rmh TNF-α抑制胃癌细胞系MKN 45的模型中,可能是通过下调NF-κB p65的基因表达及抑制NF-κB p65蛋白的核转位来抑制肿瘤增殖。
Objective To observe the changes of NF-κB p65 gene and protein expression in rmh TNF-α-MKN45 human gastric cancer cell line model. METHODS: Human gastric cancer cell line MKN45 was treated with different concentrations of rmh TNF-α (0 IU/ml, 50 IU/ml, 100 IU/ml, 200 IU/ml). CCK-8 assay was used to determine the inhibition of cell proliferation. Nested RT-PCR The level of expression of NF-κB p65 gene was detected by chain reaction (RT-PCR) and statistical analysis was performed. Differences among groups were analyzed by one-way ANOVA. T-test was used in the comparison. The nuclear translocation of NF-κB p65 protein in gastric cancer cell line MKN45 was detected by immunofluorescence cytochemistry (IF). Results The results of CCK-8 showed that the inhibition rate of cell proliferation increased with the increase of TNF-α concentration, and the difference was statistically significant. RT-PCR results showed that with the increase of TNF-α concentration of rmh, NF-κB p65 mRNA. The expression level decreased, and the difference between groups was statistically significant. IF results showed that the nuclear activity of NF-κB p65 protein decreased with the increase of rmh TN concentration. Conclusion In the model of rmh TNF-α inhibiting gastric cancer cell line MKN 45, tumor proliferation may be inhibited by down-regulating NF-κB p65 gene expression and inhibiting nuclear translocation of NF-κB p65 protein.