苦参碱对大肠癌细胞增殖和表达增殖诱导配体的影响

Effects of matrine on expression of a proliferation-inducing ligand in colorectal cancer cell lines

目的通过研究苦参碱对大肠癌细胞株SW480增殖和表达增殖诱导配体（a proliferationinducing ligand，APRIL）水平的影响，为苦参碱和APRIL的抗癌研究提供理论基础。方法采用MTT法检测苦参碱对大肠癌细胞株SW480增殖的抑制率。免疫组化法和实时荧光定量PCR检测大肠癌SW480细胞株的APRIL表达，用实时荧光定量PCR检测加入不同终浓度（0．5、1．0、2．0mg／ml）的苦参碱后24、48、72h SW480细胞表达APRIL mRNA的水平，设氟尿嘧啶为对照组及不加药为空白对照组。结果苦参碱对大肠癌SW480细胞的增殖有明显的抑制作用，并且呈剂量和时间依赖。SW480细胞有明显的APRIL表达，加入不同浓度（50、100、200μg／ml）的氟尿嘧啶后，8W480细胞APRIL mRNA的水平均逐渐升高，至72h表达最高并与空白对照组相比差异均有统计学意义（P〈0．001），而加入不同浓度苦参碱，在药物作用后24h，SW480细胞的APRIL mRNA水平升高并与空白对照组相比差异有统计学意义（P〈0．001），而后逐渐下降，至药物作用后72h，其水平与空白对照组间差异无统计学意义（P〉0．05）。结论氟尿嘧啶在抑制SW480细胞增殖的同时，残存癌细胞的增殖能力有可能因为APRIL mRNA水平的高表达而增强，故同时辅予对抗APRIL功能的治疗措施可能具有潜在的重要作用，而苦参碱不会引起SW480细胞APRIL mRNA水平的持续升高，可望成为一个有益的抗大肠癌辅助治疗手殷。

Objective To study the effect of matrine on the expression of a proliferation inducing ligand （APRIL） in colorectal cancer cell line （SW480 cell）. Methods MTT assay was used to evaluate the inhibitory effect of matrine on SW480 cells. The protein and mRNA levels of APRIL in SW480 cells were determined by immunohistochemistry and real time fluorescence quantitative PCR （RFQ PCR）. SW480 cells were treated with 0.5,1.0,2.0 mg/ml of matrine for 24 h, 48 h and 72 h. FU and blank were served as drug control and blank control groups, respectively. Results Matrine had obviously inhibitory effect on proliferation of SW480 cells in a time and dose-dependant manner. The expression of APRIL was strong in SW480 cells. When treated with 50,100,200 μg/ml of FU, the APRIL mRNA levels in SW480 cells raised gradually and reached the highest levels at 72 h after treatment, which were significantly higher than those in blank control group （all P value〈0. 001）. When treated with 0.5,1.0, 2.0 mg/ml of matrine, the APRIL mRNA levels in SW480 cells increased at 24 h after treatment, which were significantly higher than those in blank control group （all P value〈0. 001）, and then decreased gradually and almost equal to level of blank control group at 72 h. Conclusion In treatment with FU, the survival cells may have stronger ability of proliferation due to higher expression of APRIL in SW480 cells. Anti-APRIL therapy might be an important assistant treatment to counter the impact of APRIL. Matrine will not cause persistent increase of APRIL mRNA levels in SW480 cells, so it might be a helpful drug in anti-tumor theraphy.