【摘要】 目的:探讨胃上皮永生细胞、胃腺癌细胞和胃腺癌淋巴结转移细胞中多耐药基因1(mdr1)和P糖蛋白(P gp)的表达。方法:常规培养胃上皮永生细胞系(GES-1)、胃腺癌细胞系(BGC823)和胃腺癌淋巴结转移细胞系(SGC7901)细胞,用RT PCR和原位杂交检测此3种细胞的mdr1mRNA表达,用免疫印迹和免疫组化检测它们的P gp表达,用流式细胞仪检测阿霉素在细胞内的蓄积以判断它们的P gp功能状态。结果:在原位杂交的标本上,杂交信号呈紫蓝色颗粒,分布于胞质。GES1细胞颗粒数量少,BGC823颗粒数量多,细胞轮廓十分明显;SGC7901颗粒细小而少,少数细胞杂交信号丰富。免疫印迹显示GES1P gp的表达较BGC823和SGC7901弱;GES1P- gp积分光密度为46.36±19.24,BGC823为61.32±28.46,SGC7901为50.45±21.36,BGC823的P gp表达强于SGC7901和GES1(P<0.05),SGC7901和GES1的差异无统计学意义(P>0.05)。GES1阿霉素特异荧光强度为10.12±4.18,BGC823为27.44±7.06,SGC7901为35.88±14.55,3者相比差异有统计学意义(P<0.001)。结论:在胃癌的发生发展中,细胞对抗肿瘤药的反应性不同,随着细胞恶性变,P gp介导的耐药性增加。更多还原

【Abstract】 Aim: To explore expression of multidrug resistance gene 1 and P-glycoprotein in GSE-1 cell, BGC823 cell, and SGC7901 cell. Methods:GES-1(the immortal cell line), BGC823(the gastric adenocarcinoma cell line) and SGC7901(the gastric adenocarcinoma lymph node metastasis cell line) were cultured in RPMI1640 routinely. The expression of mdr1 mRNA in these cell lines was detected respectively by RT-PCR and hybridization in situ. The P-gp expression was detected by immunoblot and immunohistochemistry. The fluorescence intensity of intracellular adriamycin was assessed by flow cytometry to demonstrate the P-gp function state. Results: The hybridization signals in violet colored granules were dispersed in the cytoplasm. Based on the immunoblot, the expression level of P-gp in the GES-1 cells was lowest. The OD integrals were 46.36±19.24,61.32±28.46,50.45±21.36 in the GES-1,BGC823,SGC7901 cells, respectively. The expression level of P-gp in the BGC823 cells was stronger than those of SGC7901 and GES-1 cells(P<0.05);while there was no significant difference between SGC7901 and GES-1 cells(P>0.05).The values of adriamycin-specific fluorescence intensity were 10.12±4.18,27.44±7.06,35.88±14.55 in the GES-1,BGC823 and SGC7901 cells, respectively(P< 0.001).Conclusion: It is suggested that the cell response to antitumor drug may be different during occurrence and development of the gastric cancer, and P-gp mediated multidrug resistance may be increased with the cell malignancy.更多还原