摘要
目的探讨肿瘤坏死因子(TNF-α)诱导胃癌细胞侵袭力的相关机制。方法体外培养人胃癌细胞株SGC-7901,采用免疫组化方法检测其在TNF-α不同浓度、时间诱导下的NF-κB活性、uPA蛋白表达强度;采用噻唑蓝(MTT)比色法检测SGC-7901细胞在不同浓度的TNF-α、PDTC(吡咯烷二硫代氨基甲酸盐,NF-κB特异性抑制剂)孵育下的细胞增殖情况;构建侵袭小室,检测不同浓度TNF-α对SGC-7901细胞侵袭力影响,并用PDTC预先处理细胞后,检测其对细胞侵袭力的改变。结果随着TNF-α浓度的增加和时间延长,SGC-7901细胞NF-κB活性明显增强,90 min为高峰;TNF-α明显上调SGC-7901细胞中的uPA蛋白表达水平;一定范围浓度的TNF-α、PDTC对SGC-7901细胞增殖无明显影响;随着TNF-α浓度增加SGC-7901细胞侵袭数也随之增加,具有明显浓度依赖性(TNF-α5、10、20μg/L vs 0μg/L,84.33±3.786、108.33±6.110、121.330±4.163 vs 63.330±4.933,F=126.282,P<0.001),PDTC明显抑制TNF-α诱导的SGC-7901细胞侵袭力(42.330±4.041 vs 63.330±4.933,F=126.282,P<0.05)。结论TNF-α能够增强胃癌细胞的侵袭力,可能是通过NF-κB信号转导路径,激活uPA等蛋白水解酶而实现的。
Objective To explore the mechanism of tumor necrosis factor-alpha（TNF-α） in the promotion of human gastric carcinoma cell invasion. Methods By using immunohistochemical technique to detect the activity of nuclear factor-kappa B （NF-kB） and the expressions of urokinase type plasminogen activators （uPA） in SGC-7901 cells treated with different concentrations of TNF-α. The respective effect of TNF- α and pyrrolidine dithiocarhamate（ PDTC, an inhibitor of NF-kB） on gastric cancer cell proliferation was detected with MTT. After pretreatment with PDTC and treated with different concentrations of TNF-α, the invasion of SGC-7901 cells was observed by Boyden migration assay. Results Immunohistochemistry assay showed that the activity of NF-kB and the expression of uPA were significantly increased after TNF-α treatment. TNF-α and PDTC did not affect the proliferation of SGC-7901 cells in limited concentrations, furthermore TNF-α treatment resulted in increased invasions of SGC-7901 cells in dose-dependent and time-dependent manner （ cell-number as TNF-α at 5,10,20 ug/L vs 0 ug/L ： 84.33 ± 3. 786, 108.33 ± 6.110, 121. 330 ± 4. 163 vs 63. 330 ± 4. 933. F = 126. 282. P 〈 0. 001 ）. while TNF-α induced invasion of SGC-7901 cells was markedly inhibited by pretreatment with PDTC （ cell number ： 42. 330 ± 4. 041 vs 63. 330 ± 4. 933. F = 126. 282. P 〈 0.05）. Conclusions TNF-α promotes the invasion of human gastric carcinoma cells, and the up-regulation of uPA and NF-kB activity is involved in this proce±ss.
出处
《肿瘤基础与临床》
2008年第3期202-205,共4页
journal of basic and clinical oncology
关键词
肿瘤坏死因子
尿激酶型纤溶酶原激活物
核转录因子-KB
胃癌
肿瘤浸润
tumor necrosis factor-alpha
urokinase type plasminogen activator
nuclear factor-kappa B
gastric carcinoma： neoplasm invasiveness
