吗啡对人肝癌SMMC-7721细胞PTCH-1、GLI-2和MMP-9 mRNA表达的影响

Effect of morphine on PTCH-1,GLI-2 and MMP-9 mRNA expression in human hepatocellular carcinoma SMMC-7721 cell

目的观察吗啡对肝癌SMMC-7721细胞迁移能力及丁蛋白受体1(PTCH-1)、胶质瘤相关癌基因同源蛋白2(GLI-2)、基质金属蛋白酶9(MMP-9)mRNA表达的影响。方法将含1μmol/L(M1组)、10μmol/L(M2组)、100μmol/L(M3组)吗啡的细胞培养基,分别孵育SMMC-7721细胞48h,空白对照组(C组)不加入药物干预,每组6孔。应用划痕实验观察细胞迁移能力的变化,应用半定量逆转录-聚合酶链反应(RT-PCR)检测肝癌SMMC-7721细胞中PTCH-1、GLI-2和MMP-9mRNA的表达。结果划痕损伤48h后,M1组、M2组和M3组细胞的愈合率均明显低于C组,细胞的迁移能力受到明显抑制(P<0.05);RT-PCR结果显示,吗啡使M1组、M2组和M3组PTCH-1mRNA表达均明显高于C组(P<0.05);GLI-2mRNA表达均较C组明显下调,M1组表达明显低于C组(P<0.05),而M2组和M3组差异无统计学意义;MMP-9mRNA在M1组表达明显低于C组(P<0.05),M2组和M3组表达虽高于C组,但差异无统计学意义。结论吗啡能抑制肝癌细胞SMMC-7721的迁移能力,其机制可能与抑制Hedgehog信号通道活性并诱导下调MMP-9的表达相关。

Objective To explore the effects of morphine on the migration ability of human hepatocellular carcinoma SMMC-7721 cells and the expression of PTCH-1,GLI-2and MMP-9 mRNA.Methods SMMC-7721 cells were incubated for 48 hours in different mediums containing 1μmol/L（group M1）,10μmol/L（group M2）and 100μmol/L（group M3）morphine respectively.Medium without morphine as negative control.Each group included six holes.The wound healing assay was used to detect the migration activity of SMMC-7721 cell.The expression of MMP-9,PTCH-1and GLI-2mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction（RTPCR）.Results The healing rates of all morphine treatment groups were lower than that of the control group（P〈0.05）.PTCH-1 mRNA up-regulation and GLI-2 mRNA down-regulation were found in morphine groups（P〈0.05）;MMP-9mRNA down-regulated in 1μmol/L morphine group,otherwise up-regulated in 10 and 100μmol/L morphine groups（P〈0.05）.Conclusion Morphine would inhibit migration ability of hepatocellular carcinoma SMMC-7721 cell.The mechanism might be related to hedgehog signaling pathway activity inhibition and the reduced expression of MMP-9.