重楼提取液对人结肠癌SW480细胞增殖的影响及其作用机制  

Effect of Paridis Extract on Proliferation of Human Colon Cancer SW480 Cells and Mechanism of The Effect

作　　者：李晞[1,2] 王继红[1,2] 肖亚雄[1,2] 

机构地区：[1]重庆医科大学基础医学实验教学中心生物化学与分子生物学实验室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国生物制品学杂志》2010年第6期619-622,632,共5页Chinese Journal of Biologicals

基　　金：重庆市科委自然科学基金(CSTC;2008CE9142);重庆医科大学创新基金(CX200528)

摘　　要：目的观察重楼提取液对人结肠癌SW480细胞增殖的影响,并探讨其作用机制。方法采用不同浓度(10、20、40、80μg/ml)的重楼提取液,分别作用于体外培养的SW480细胞12、24、36和48h,采用MTT法检测其对SW480细胞的抑制率;并以半数抑制浓度的重楼提取液与体外培养的SW480细胞作用24h后,显微镜下观察细胞形态,并通过流式细胞术分析其对SW480细胞细胞周期的影响,RT-PCR和Western blot法分别测定SW480细胞干细胞因子(SCF)mRNA和蛋白表达的变化。结果各浓度重楼提取液对SW480细胞均有不同程度的抑制作用,且存在剂量-效应和时间-效应关系,重楼提取液作用后的SW480细胞出现变性、坏死的形态改变;经重楼提取液处理24h后,SW480细胞在S期的分布比例下降,G0-G1期和G2/M期细胞分布增多(P<0.05);同时SCF表达增高(P<0.05)。结论重楼提取液可能通过抑制肿瘤细胞的蛋白质与DNA合成,抑制肿瘤细胞的有丝分裂,进而抑制SW480细胞增殖,且其作用机制可能与SCF无关。Objective To observe the effect of Paridis extract on proliferation of human colon cancer SW480 cells and investigate the mechanism of the effect.Methods SW480 cells cultured in vitro were treated with various concentrations（10,20,40 and 80 μg/ml）of Paridis extract for 12,24,36 and 48 h separately.The inhibiting rate to SW480 cells was determined by MTT method.The SW480 cells cultured in vitro were treated with Paridis extract at median inhibitory concentration for 24 h,then observed for morphology and analyzed the effect on cell cycle by flow cytometry.The transcription of stem cell factor（SCF）mRNA in SW480 cells was determined by RT-PCR,and expression of SCF protein by Western blot.Results The Paridis extract at various concentrations showed inhibitory effect at different degrees on SW480 cells,in dose-and time-dependent modes.Morphological change of SW480 cells,such as denaturation and necrosis,were observed after treatment.The percentage of SW480 cells at S phage decreased 24 h after treatment with Paridis extract,while those at G0-G1 and G2 /M phases increased（P〈0.05）.Meanwhile,the expression level of SCF increased（P〈0.05）.Conclusion Paridis extract might inhibit the mitosis of tumor cells by inhibiting the synthesis of protein and DNA,thereby inhibit the proliferation of SW480 cells,and the inhibitory effect might be SCF-independent.

关 键 词：重楼 结肠肿瘤 细胞增殖 干细胞因子 

分 类 号：R735.3[医药卫生—肿瘤]