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目的探讨pokemon基因表达与胃癌细胞药敏性及耐药基因的关系。方法取60例胃癌组织和癌旁组织石蜡标本,采用免疫组化技术检测组织中pokemon蛋白及耐药相关蛋白P-糖蛋白(P-gp)、多药耐药相关蛋白1(MRP1)、肺耐药蛋白(LRP)、生存素(survivin)的表达,并分析pokemon蛋白与其他4种蛋白的关系。采用磺酰罗丹明B(SRB)蛋白染色法检测5-氟尿嘧啶(5-FU)、顺铂(DDP)、奥沙利铂(L-OHP)对胃癌细胞株SGC7901、胃癌耐药细胞株SGC7901/ADR、胃上皮细胞株GES-1的抑制作用;qPCR及蛋白质印迹法检测上述3种细胞株中pokemon和耐药基因的表达。合成针对pokemon的siRNA并转染SGC7901/ADR细胞,检测转染前后SGC7901/ADR细胞的化疗药敏性及耐药基因的变化。结果胃癌组织中pokemon、P-gp、MRP1、LRP、survivin蛋白的阳性表达率均高于癌旁组织(P<0.05);pokemon蛋白表达与P-gp、survivin蛋白表达间存在正相关关系(r=0.385 2,P=0.002;r=0.342 4,P=0.007)。Pokemon、P-gp、MRP1、survivin蛋白在SGC7901/ADR细胞株中的表达高于细胞株SGC7901及GES-1;SGC7901/ADR细胞株的耐药性最强,SGC7901次之,GES-1最弱(P<0.01)。Pokemon-siRNA转染SGC7901/ADR细胞后细胞中pokemon mRNA的表达明显受到抑制;转染后化疗药物对细胞的抑制能力增强,P-gp、survivin表达减弱(P<0.05)。结论 Pokemon与P-gp、survivin通过相互调控而促进胃癌耐药性的形成。Pokemon可能成为胃癌靶向治疗的候选基因。
Objective To investigate the relationship between pokemon gene expression and drug sensitivity and drug resistance genes in gastric cancer cells. Methods Sixty paraffin-embedded specimens of gastric cancer tissues and para-cancerous tissues were collected. Immunohistochemistry was used to detect the expression of pokemon protein and P-glycoprotein (P-gp), multidrug resistance-related protein 1 (MRP1) The expression of LRP and survivin was analyzed and the relationship between pokemon and other four proteins was analyzed. Gastric cancer cell line SGC7901, gastric cancer cell line SGC7901 / SGC7901 were treated with 5-fluorouracil (5-FU), cisplatin (DDP) and oxaliplatin (L-OHP) ADR and gastric epithelial cell line GES-1. The expression of pokemon and drug resistance gene in the three cell lines was detected by qPCR and Western blot. The siRNA targeting pokemon was synthesized and transfected into SGC7901 / ADR cells. The chemosensitivity and drug resistance genes of SGC7901 / ADR cells were detected before and after transfection. Results The positive rates of pokemon, P-gp, MRP1, LRP and survivin in gastric cancer tissues were significantly higher than those in adjacent tissues (P <0.05). There was a positive correlation between the expressions of P-gp and survivin protein = 0.385 2, P = 0.002; r = 0.342 4, P = 0.007). The expression of Pokemon, P-gp, MRP1 and survivin in SGC7901 / ADR cell lines was higher than that in SGC7901 and GES-1 cell lines; SGC7901 / ADR cell lines were the most resistant cells, followed by SGC7901 and GES- 1 (P <0.01). Pokemon-siRNA transfected SGC7901 / ADR cells in the pokemon mRNA expression was significantly inhibited; transfected cells after chemotherapy drugs inhibit the ability of P-gp, survivin expression decreased (P <0.05). Conclusion Pokemon and P-gp, survivin promote the formation of gastric cancer drug resistance by mutual regulation. Pokemon may be a candidate gene for gastric cancer targeted therapy.