隐丹参酮对胰腺癌细胞生物学行为的影响及作用机制

Influence of cryptotanshinone on biological behaviors of pancreatic cancer cells and the action mechanism

背景与目的:胰腺癌是一种高度恶性的消化系统肿瘤,尽管目前针对胰腺癌的治疗方式在不断发展,患者预后仍然很差。隐丹参酮(CPT)是从植物丹参中提取的具有多种活性的单体,已被证实对宫颈癌、前列腺癌等有抗癌作用,但其对胰腺癌的作用尚不清楚,本研究通过体外实验探讨CPT对胰腺癌细胞生长和迁移的影响及相关机制,为相关的临床治疗药物研发提供理论和实验依据。方法:采用人胰腺癌BxPC-3细胞和SW1990细胞为研究对象,用3个浓度(10、20、40μmol/L)的CPT处理该两种胰腺癌细胞不同时间(0、1、2、3 d)后,用CCK-8法检测CPT对细胞活力影响的浓度与时间效应;用以上3个浓度的CPT处理两种胰腺癌细胞24 h后,分别用克隆形成实验、划痕实验、Transwell实验检测对细胞克隆形成能力、迁移能力、侵袭能力的变化;用20μmol/L CPT处理BxPC-3和SW1990细胞1、2、3 d后,用Western blot检测Akt、磷酸化Akt(p-Akt)以及上皮-间质转化(EMT)相关蛋白vimentin、E-cadherin与细胞周期相关蛋白CDK4、cyclin D1的表达。对照组细胞均只采用溶剂(DMSO)处理。结果:与各自对照组比较,CPT处理后的两种胰腺癌细胞的生长被明显抑制,且呈时间与浓度依赖性(均P<0.05);划痕愈合程度、相对克隆形成率、侵袭细胞数均明显降低,且呈浓度依赖性(均P<0.05);Akt、p-Akt、vimentin、E-cadherin、CDK4、cyclin D1蛋白的表达均明显下调,且呈时间依赖性(均P<0.05)。结论:CPT能有效抑制胰腺癌细胞的生长和迁移,其作用机制可能与其下调Akt的表达,并进一步导致胰腺癌细胞生长周期阻滞及抑制EMT过程有关。

Background and Aims:Pancreatic cancer is a highly malignant tumor of the digestive system.Although the current treatment for pancreatic cancer is constantly evolving,the prognosis of pancreatic cancer patients is still poor.Cryptotanshinone(CPT)is a monomer extracted from Chinese medicinal herb salvia miltiorrhiza with variety of activities,which has been proven to have anti-cancer potential against cervical cancer and prostate cancer,etc.However,its effect on pancreatic cancer is still unclear.This study was conducted to investigate the effect of CPT on the growth and migration of pancreatic cancer cells in vitro and the possible mechanism,so as to provide theoretical and experimental basis for the development of relevant drugs for clinical use.Methods:The human pancreatic cancer BxPC-3 and SW1990 cells were used as study subjects.In these two types of pancreatic cancer cells,the concentration and time effects of CPT on cell viability were determined after treatment with different concentrations(10,20 and 40μmol/L)of CPT for different times(0,1,2 and 3 d);the changes in abilities of colony formation,migration and invasion after treatment with above concentrations of CPT for 24 h were determined by colony forming assay,scratch wound healing assay and Transwell assay,respectively;the expressions of Akt,phosphorylated Akt(p-AKT),and the epithelial-mesenchymal transition(EMT)-associated proteins vimentin and E-cadherin as well as the cell cycle-related proteins CDK4,cyclin D1 after treatment with 20μmol/L CPT for different times were determined by Western blot analysis.The cells for control were treated with the vehicle(DMSO)only.Results:In the two types of pancreatic cancer cells compared with corresponding control group,the growth abilities were significantly inhibited after CPT treatment,with a concentration-and time-dependent manner(all P<0.05);the degree of wound healing,relative colony formation rate and number of invading cells were significantly reduced,with a concentration-dependent manner(all P<0.05);the protein expressions of Akt,p-Akt,vimentin and E-cadherin,CDK4 and cyclin D1 were all significantly down-regulated,with a time-dependent manner(all P<0.05).Conclusion:CPT can effectively inhibit the growth and migration of pancreatic cancer cells,and the mechanism may be probably associated with its downregulating the expression of Akt and thereby,cause the cell cycle arrest and inhibition of the EMT process of the pancreatic cancer cells.