Hedgehog信号通路相关蛋白在人胰腺癌SW1990耐药株中的表达及意义

Expression and functional significance of hedgehog signal pathway members in drug-resistant SW1990 pancreatic cancer cell line

目的:探讨Hedgehog信号通路蛋白在人胰腺癌吉西他滨耐药细胞株SW1990中的表达,为克服胰腺癌获得性耐药提供实验基础。方法:采用浓度梯度递增法建立人胰腺癌SW1990耐药株,采用噻唑蓝法测定SW1990亲代与耐药细胞IC50。实时荧光定量PCR检测亲代与耐药细胞mRNA中hedgehog信号通路成员Shh、SMO、Gli-1的表达差异。Western印迹法检测亲代与耐药细胞中上述蛋白质的表达。结果:人胰腺癌耐药株SW1990的IC50从亲代的(3.1±0.2)μmol/L提高到(232.2±12.3)μmol/L。荧光定量PCR结果显示耐药株中Shh、Gli-1的表达提高了(12.07±1.71)倍和(4.15±0.42)倍。亲代SW1990中未检测到SMO表达,而耐药细胞中却可以检测到SMO的表达。Western印迹结果同样显示,人胰腺癌SW1990耐药细胞株中高表达上述蛋白质。结论:人胰腺癌耐药株中高表达部分hedgehog信号通路蛋白。针对hedgehog信号通路的靶向治疗可能为克服胰腺癌耐药提供新的理论基础。

Objective To investigate the difference of protein expression in hedgehog signal pathway in parental and gemeitabine-resistant pancreatic cancer cell line SW1990. Methods Gemcitabine-resistant cell line SW1990 was established by drug concentration step-elevation method. IC50 to gemeitabine in parental and gemeitabine-resistant cells were determined by MTT. mRNA expression levels of Shh, SMO and Gli-I were determined by real-time PCR. Protein expression level of Shh, SMO and Gli-1 were determined by Western blot. Results ICs0 to gemcitabine was （232.2± 12.3） μmol/L in drug-resistant cells and （3.1-＋0.2） μmol/L in parental cells, mRNA expression of Shh and Gli-1 in gemcitabine-resistant cells increased （12.07±1.71） and （4.15±0.42） fold, respectively, when compared with that in parental cells, mRNA expression of SMO was detectable only in gemcitabine-resistant cells, but not in parental cells. Western blot showed a high expression levels of SMO and Gli-1 proteins in gemcitabine-resistant cells. Conculsions Some members of hedgehog signal pathway are highly expressed in gemcitabine-resistant pancreatic cancer cells. Targeting therapy against hedgehog pathway may contribute to overcome gemcitabine-resistant in pancreatic cancer.