摘要
目的 :研究甲基莲心碱 (Nef)逆转耐长春新碱人胃癌细胞多药耐药性 (MDR)的作用及机制。方法 :采用噻唑蓝 (MTT)比色法检测长春新碱 (VCR)的细胞毒性 ;PI染色流式细胞计数测定VCR诱导细胞凋亡 ;间接免疫荧光流式细胞术检测细胞P -gp和MRP的表达。结果 :Nef (5、10 μmol·L-1)对人胃癌细胞 (SGC790 1)和耐长春新碱人胃癌细胞 (SGC790 1/VCR)无显著毒性作用 ,VCR对敏感株SGC790 1的IC50 为 0 0 6mg·L-1,而对MDR细胞株SGC790 1/VCR的IC50 为 2 32mg·L-1,SGC790 1/VCR较SGC790 1对VCR耐药 39倍 ,Nef(2 5、5、10 μmol·L-1)能使VCR对SGC790 1/VCR细胞的IC50 从 2 32mg·L-1依次下降至 0 34、0 12、0 0 5mg·L-1,逆转倍数分别为 6 8、18 1、4 3 8。Nef(2 5、5、10 μmol·L-1)能降低SGC790 1/VCR细胞对VCR的凋亡抗性 ,其作用强于维拉帕米 (VRP)。SGC790 1/VCR细胞较SGC790 1细胞高表达P -gp、MRP ,Nef(10 μmol·L-1)处理 2 4h后 ,SGC790 1/VCR细胞P -gp、MRP的表达明显低下。结论 :Nef具有逆转耐长春新碱人胃癌细胞的MDR作用 ,其作用机理与下调P
AIM: To investigate the effect of neferine (Nef) on human gastric carcinoma cell line with multidrug resistance (MDR). METHODS: The cytotoxic effect of vincristine (VCR) was evaluated by MTT assay. The cell apoptosis induced by VCR was determined by flow cytometry, and the expression of P-glycoprotein (P-gp) and multidrug-resistance-associated protein (MRP) in cells was examined by immunofluorescence flow cytometry. RESULTS: MTT assay showed that Nef at the concentration of 5 μmol·L^(-1) to 10 μmol·L^(-1) have no cytotoxicity to parent human gastric carcinoma cell line (SGC7901) and its VCR-resistant variant cell line (SGC7901/VCR). The IC_(50) value of VCR to SGC7901 cell line was 0.06 mg·L^(-1)and that of to SGC7901/VCR cell line was 2.32 mg·L^(-1), which indicated SGC7901/VCR cell line were 39 times more resistant to VCR in comparison with the parent SGC7901 cell line. After treatment with Nef at the concentrations of 2.5, 5 and 10 μmol·L^(-1), the IC_(50) value of VCR to SGC7901/VCR cell line decreased to 0.34, 0.12 and 0.05 mg·L^(-1), respectively and those increased by 6.8-, 18.1- and 43.8- fold in the chemosensitivity, respectively. Flow cytometry showed that SGC7901/VCR cells were resistant to apoptosis induced by VCR. After 24 h treatment with Nef (2.5, 5 and 10 μmol·L^(-1)) and VCR, the apoptosis of SGC7901/VCR cells increased, which indicated Nef could abolish resistance of SGC7901/VCR cells to VCR-induced apoptosis. Furthermore, the action of Nef was more potent than verapamil. The expression of P-glycoprotein and multidrug resistance associated protein was strongly positive in SGC7901/VCR cells, and the expression level of P-gp and MRP in SGC701/VCR cells was significantly down-regulated at 24 h after treatment with Nef (10 μmol·L^(-1)). CONCLUSIONS: Nef can reverse MDR in multidrug-resistant human gastric carcinoma SGC7901/VCR cell line. Its mechanism might be associated with down-regulation of expression of P-gp and MRP in SGC7901/VCR cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第8期1407-1410,共4页
Chinese Journal of Pathophysiology
基金
湖南省教育厅科研基金资助课题 (No .96 15 5 )
