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目的 为研究环氧合酶 2 (cyclooxygenase 2 ,COX 2 )或通过前列腺素 ( prostaglandin ,PG)引起的肿瘤免疫机制 ,我们检测了人胃癌细胞中COX 2mRNA及蛋白质水平的表达 ,PGE2的释放和COX 2酶活性的抑制剂消炎痛 (in domethacin ,Indo)对PGE2的释放和肿瘤细胞生长的影响。方法 采用免疫细胞化学染色、RT PCR ,ELISA ,以及MTT比色试验方法研究人胃癌细胞COX 2基因与蛋白质的表达、PGE2释放以及消炎痛对PGE2释放和细胞活力的影响。结果 所有检测的胃癌细胞均表达COX 2mRNA ,胃癌细胞MKN45 ,SGC790 1和AGS中有COX 2蛋白质的表达 ,MGC80 3则不表达。ELISA表明 ,所有胃癌细胞均有PGE2释放 ,给予COX 2表达的诱导剂PMA后 ,MGC80 3中PGE2的释放由 5 7μg/L增加至 6 3μg/L ,小剂量COX 2酶活性的抑制剂—消炎痛 ( 0 12 5 μmol/L)则可使其水平降至 47μg/L。MTT法进一步显示 ,消炎痛可使肿瘤细胞增殖与活力明显抑制 ,尤以小剂量为著。结论人胃癌细胞中存在COX 2基因的表达、PGE2的释放及肿瘤细胞增殖之间的相互关联
Objective To investigate the immune mechanisms of cyclooxygenase 2 (COX 2 ) or tumors induced by prostaglandin (PG), we examined the expression of COX 2 mRNA and protein levels in human gastric cancer cells, and the release of PGE 2 and COX. 2 Inhibitory activity of indothacin (Indotha), an inhibitor of enzyme activity, on PGE2 release and tumor cell growth. Methods Immunocytochemical staining, RT PCR, ELISA, and MTT colorimetric assay were used to investigate the effects of COX 2 gene and protein expression, PGE 2 release, and indomethacin on PGE 2 release and cell viability in human gastric cancer cells. Results COX 2 mRNA was expressed in all gastric cancer cells. COX 2 protein was expressed in gastric cancer cells MKN45, SGC790 1 and AGS, but not in MGC80 3. ELISA showed that all gastric cancer cells had PGE2 release. After administration of PMA, an inducer of COX 2 expression, the release of PGE2 in MGC80 3 increased from 57 μg/L to 63 μg/L. The inhibitor of low-dose COX 2 enzyme activity was anti-inflammatory. Pain ( 0 12 5 μmol/L) reduced the level to 47 μg/L. The MTT method further showed that indomethacin can significantly inhibit the proliferation and viability of tumor cells, especially at low doses. Conclusion There is a correlation between COX 2 gene expression, PGE 2 release and tumor cell proliferation in human gastric cancer cells.