氧化苦参碱对人胰腺癌细胞株SW1990侵袭转移能力的影响

目的 探讨氧化苦参碱对人胰腺癌细胞株SW1990体外迁移及侵袭能力的影响及其作用机制.方法 体外培养人胰腺癌SW1990细胞株,用氧化苦参碱处理SW1990细胞后,采用MTT法检测细胞增殖;通过细胞黏附实验、细胞划痕实验及Transwell小室检测细胞的黏附、迁移及侵袭能力;RT-PCR法检测细胞基质金属蛋白酶2(MMP-2)、血管内皮生长因子(VEGF)mRNA的表达;ELISA法检测细胞VEGF蛋白的含量.结果 氧化苦参碱呈剂量和时间依赖性抑制SW1990细胞的增殖.2 mg/ml氧化苦参碱处理SW1990细胞1 h后,细胞的体外黏附抑制率为(35.23 ±8.56)%;处理24 h后,细胞的过河时间为(65.46±4.25)h,较对照组的(34.50±4.12)h显著延长(P<0.05);穿膜细胞数为(91.9±9.6)个,较对照组的(144.2±17.2)个显著减少(P<0.05);细胞VEGF、MMP-2 mRNA的表达及VEGF蛋白的分泌量均显著下调0.515 ±0.063比0.817±0.054,0.343±0.072比0.650±0.068,(265.50 ±5.45)pg/ml比(441.06±16.70)pg/ml,P值均<0.05].结论 氧化苦参碱可能通过抑制MMP-2和VEGF表达进而抑制胰腺癌SW1990细胞的增殖、黏附、迁移及侵袭能力.

Influence of oxymatrine on metastatic and invasive ability of human pancreatic cancer cell line SW1990

Objective To investigate the effect and mechanism of oxymatrine on invasion and metastasis of human pancreatic cancer line SW1990 in vitro. Methods Human pancreatic cancer cell line SW1990 was cultured in vitro. Oxymatrine was added into the culture media of SW1990 cells. Then MTT assay was used to determine the effect on proliferation. The adhesive capability, the mobile ability and invasive ability of SW1990 cells were detected by the adhesion assay, the crossing-river test, the transwell migration assay and the matrigel invasion method, respectively. RT-PCR was used to detect the mRNA expression of MMP-2 and VEGF. ELISA method was used to detect the protein levels of the VEGF. Results The growth of SW1990 cells was inhibited by oxymatrine in a dose and time-dependent manner. After 2 mg/ml of oxymatrine treatment for SW1990 cells for 1 h, the adhesive capability inhibitory rate was (35.23 ±8.56) % ; 24 h later, crossing-river time was (65.46 ±4.25) h, which was significantly longer than that in control group (34.50 ± 4.12) h, P <0.05)], the number of penetrating cells was 91.9 ±9.6, which was significantly lower than that in control group (144.2±17.2, P <0.05). The mRNA expression of MMP-2 and VEGF, expression of protein of VEGF in SW1990 cells was significantly down-regulated 0.515 ±0.063 vs. 0.817 ±0.054, 0.343 ± 0.072 vs. 0.650 ±0.068; (265.50 ±5.45) pg/ml vs. (441.06 ±16.70) pg/ml, P <0.05]. Conclusions Oxymoron can inhibit the invasion and metastasis ability of pancreatic cancer line SW1990 in vitro, and the mechanism is possibly related to the down-regulation of MMP-2 and VEGF expression.