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目的:研究复方五味子素B(γSC)对耐奥沙利铂人结肠癌细胞(THC-8307/OXA)多药耐药性的逆转作用及作用机制。方法:采用噻唑蓝(MTT)比色法检测奥沙利铂(OXA)和γSC的细胞毒性;碱性磷酸酶免疫组织化学法和Western blot检测γSC对THC-8307/OXA细胞GST-п蛋白水平的影响。结果:γSC(6.0,12.5,25μg·ml~(-1))对人结肠癌细胞(THC-8307)和THC-8307/OXA无显著毒性作用(P>0.05),OXA对THC-8307的IC_(50)为0.06μg·ml~(-1),而对THC-8307/OXA的IC_(50)为2.32μg·ml~(-1),THC-8307/OXA较THC-8307耐药性提高39倍,γSC(6.0,12.5,25μg·ml~(-1))能使OXA对THC-8307/OXA细胞的IC_(50)从2.32μg·ml~(-1)依次下降至0.370,0.128,0.057μg·ml~(-1),逆转倍数分别为6.2,18.1,40.7倍。碱性磷酸酶免疫组织化学法和Western blot检测γSC(12.5μg·ml~(-1))处理48 h后,THC-8307/OXA细胞GST-п蛋白表达明显降低(P<0.01)。结论:γSC具有逆转耐奥沙利铂人结肠癌细胞的MDR作用,其作用机制与下调GST-п表达有关。
Objective: To investigate the reversal effect and mechanism of compound Schisandrin B (γSC) on multidrug resistance in oxaliplatin-resistant human colon cancer cells (THC-8307 / OXA). Methods: MTT assay was used to detect the cytotoxicity of oxaliplatin (OXA) and γSC; alkaline phosphatase immunohistochemistry and Western blot were used to detect the expression of GST-п protein in THC-8307 / OXA cells Impact. Results: γSC (6.0,12.5,25μg · ml -1) had no significant toxic effect on human colon cancer cells (THC-8307) and THC-8307 / OXA (P> 0.05) (50) of THC-8307 / OXA was 0.06μg · ml ~ (-1), while the IC50 of THC-8307 / OXA was 2.32μg · ml ~ (-1) 39 times, γSC (6.0,12.5,25μg · ml -1) decreased the IC 50 of OXA from 2.32μg · ml -1 to 0.370,0.128 in THC-8307 / OXA cells, 0.057μg · ml ~ (-1), the reversal fold were 6.2,18.1,40.7 times. The expression of GST-к protein in THC-8307 / OXA cells was significantly decreased after treatment with γSC (12.5μg · ml -1) for 48 h by alkaline phosphatase immunohistochemistry and Western blot (P <0.01). CONCLUSION: γSC can reverse the MDR effect of oxaliplatin-resistant human colon cancer cells, and its mechanism is related to the down-regulation of GST-п expression.