Agkihpin对人肝癌SMMC-7721细胞COX-2、MRP1和E-CD表达的影响

Effects of Agkihpin on the expression of COX-2,MRP1 and E-CD in human hepatocellular carcinoma SMMC-7721 cell strain

目的探讨蛇毒精氨酸酯酶(Agkihpin)影响人肝癌SMMC-7721细胞活力、增殖和迁移的机制,为肝癌的治疗提供新的思路。方法取对数生长期SMMC-7721细胞随机分为8组,分别用质量浓度为0.207～4.130 g/mL的Agkihpin处理72 h,应用免疫细胞化学、Western blotting、RT-PCR法检测环氧合酶-2(COX-2)、表皮钙黏素(E-CD)和多药耐药相关蛋白1(MRP1)蛋白表达和基因转录水平,分析三指标之间的关系。结果与未行Agkihpin处理者比较,Agkihpin处理的SMMC-7721细胞COX-2基因转录、蛋白表达均降低(P<0.01),并呈一定的浓度依赖效应;COX-2与E-CD的表达和转录均呈负相关,MRP1与E-CD的表达和转录均呈负相关,COX-2与MRP1的表达和转录均呈正相关。结论 Agkihpin可通过下调MRP、COX-2表达及上调E-CD表达抑制人肝癌SMMC-7721细胞的活力、增殖和迁移,有望用于提高肝癌细胞对化疗药物的敏感性、逆转肝癌耐药细胞的多药耐药表型、提高肝癌患者术后生存率、抑制肝癌细胞的浸润和转移。

Objective To explore the mechanism of Agkihpin affecting the cellular vitality, proliferation and migration of SMMC-7721, so to provide a new thinking for therapy of hepatocellular carcinoma. Methods The cultured cells growing in logarithmic growth phase were divided into 8 groups randomly, and treated with different concentrations （0. 207-4. 130 g/mL） of Agkihpin for 72 hs, then the transcription and expression levels of eyelooxygenase-2 （ COX-2 ）, multidrug resist- ance associated protein 1 （ MRP1 ） and epidermal cadherin （E-CD） in SMMC-7721 cell strain were assayed with immunocy- tochemistry, Western blotting and RT-PCR, and the relationship of these 3 indexes were analyzed. Results Compared with the Agkihpin-absented group, the transcription and expression levels of COX-2 in SMMC-7721 cell were reduced sig- nificantly after the treatment of Agkihpin （P 〈0.01 ）, in a certain concentration-dependent manner; the transcription and expression levels of both COX-2 and MRP1 showed a negative correlation with those of E-CD, while both the transcription and expression levels of MRP1 showed a positive correlation with those of COX-2. Conclusion Agkihpin may inhibit the cellular vitality, proliferation and migration via down-regulating the expressions of MRP1 and COX-2 and up-regulating the expression of E-CD of SMMC-7721. Agkihpin may be useful in therapy of hepatocellular carcinoma with enhancing the sensitivity of cancer cells to chemotherapeutic drugs, reversing the multidrug-resistantaed phenotype of hepatocellular carcinoma cell, inhibiting the soakage and migration of cancer ceils in future and increasing the hepatocellular carcinoma sufferers＇ postoperative survival rate in the future.