摘要
目的观察从丹参中提纯的单体———丹参酮ⅡA(TanⅡA)在体外对SGC7901胃癌细胞的诱导凋亡作用,并对其分子机制作初步探讨。方法分别采用不同浓度梯度(1.0、2.0、4.0、6.0、8.0和10.0mg/L)的TanⅡA干预SGC7901胃癌细胞24、48、72h后,用四唑盐(MTT)比色法绘制肿瘤细胞生长曲线。采用以上浓度梯度的TanⅡA干预SGC7901细胞24h,或采用10.0mg/L的TanⅡA干预0、12、24、36、48h,应用碘化丙叮(PI)及Annexin V双重染色法,通过流式细胞仪观察SGC7901的细胞周期及凋亡率。用透射电镜直接观察细胞凋亡形态。用RT-PCR检测不同浓度的TanⅡA干预24h后SGC7901细胞p53的mRNA表达。结果MTT法显示TanⅡA具有良好的抑制胃癌增殖作用,并时间、剂量依赖性地诱导肿瘤细胞凋亡。透射电镜下观察到典型的细胞凋亡形态,流式细胞术(FCM)显示TanⅡA可使SGC7901细胞周期阻滞于G1/S期。PCR检测发现随着TanⅡA干预浓度的逐步上升,胞内p53mRNA表达亦逐步上升。结论TanⅡA在体外可通过细胞周期阻滞及诱导凋亡达到良好的抑制胃癌细胞增殖效应,诱导胞内p53基因表达上调是所涉分子机制之一。
Purpose Tanshinone Ⅱ A is a monomer derived from Chinese traditional medicine herb Salvia miltiorrhiza Bge. The apoptosis of SGC7901 human gastric cancer cells induced by Tanshinone Ⅱ A was studied in order to assess its antitumor effect and identify the molecular mechanism involved. Methods SGC7901 gastric cancer cell line were cultured in vitro and treated by Tanshinone ⅡA of 1 - 10 μg/mL concentrations for 3 days and then the growth curves were drew based on MTT assay. Morphology and quantity of apoptosis and cell cycles were observed and analyzed by light microscopy, electronic microscopy, flow cytometry with Annexin V and propidium iodide double staining. Gene transcription of P53 was also examined by RT-PCR. Results The proliferation of SGC7901 cells was obviously inhibited by Tanshinone ⅡA and apoptosis of SGC7901 was induced by dose dependent and time dependent manner. Characteristic apoptosis were confirmed by electronic microscopy and PI and Annexin V double staining. Cell cycle was arrested into G1/S phase by Tanshinone Ⅱ A. The transcription of p53 gene showed increasing tendency when treated by 1 - 10 μg/mL Tan ⅡA for 24 hours. Conclusions Tanshinone Ⅱ A could induce apoptosis of SGC7901 gastric cancer cells by dose-dependent and time-dependent manner, which may be one of the important mechanisms of Tanshinone Ⅱ A＇s anticancer effects. P53 transcription in SGC7901 was up-regulated and cell cycle was arrested into G1/S phase by Tanshinone Ⅱ A, which may be one of the molecular mechanisms involved in the anti-cancer and proapoptotic effect of Tanshinone Ⅱ A.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2007年第1期57-61,65,共6页
Hehai University Journal of Medical Sciences
关键词
胃癌
凋亡
丹参酮
P53
gastric cancer
apoptosis
tanshinone
p53
