半边旗中二萜类化合物5F诱导胰腺癌细胞凋亡机制的探讨

Mechanisms of Inhibition of Apoptosis of Human Pancreatic cancer PC-3 cell line by a diterpenoid compound 5 F isolated from Pteris Semipinnata L treatment

目的:观察研究半边旗中二萜类化合物5F(11a-羟基-15-氧-16-烯-ent贝壳杉烷-19酸)诱导人胰腺癌细胞株PC-3凋亡的效果及其机制。方法:在体外培养的胰腺癌细胞株PC-3中加入8.875umol/L,37.5umol/L,142umol/L浓度的5F孵育24h,RT-PCR分析各组细胞中mRNA-PUMA表达,MTT检测细胞相对存活率,流式细胞仪检测细胞凋亡。利用Lipofectamine^(TM)2000将人类siRNA-PUMA转染导入胰腺癌细胞PC-3中抑制PUMA的表达后,观察转染96h后的细胞株PC-3在上述浓度的5F的作用下各组细胞中mRNA-PUMA的表达、细胞凋亡以及细胞生长的变化结果:在5F作用下,细胞mRNA-PUMA表达提高,细胞存活率随着药物浓度的提高明显降低,细胞凋亡率随着药物浓度的提高而明显提高,且呈量效关系;转染siRNA-PUMA后的细胞在5F作用下,细胞中mRNA-PUMA未见明显表达,细胞存活率和凋亡率与对照组比无显著性改变。结论:5F可能通过上调PUMA基因表达而促进肿瘤细胞凋亡,抑制肿瘤生长。

Objective：To study the effects of diterpenoid compound 5F （ent-IIa-hydrOXy-I 5-OXo-kaur-1 6-en-19-oic acid） isolated from Pteris Semipinnata L. on apoptosis of pancreatic cancer cell line PC-3,and the mechanisms involved in this effect. Methods： Human cultured pancreatic cancer cell line PC-3 was exposed in vitro to 8.875umol/L.37.5umol/L,142umol/L concentrations of 5F for 24h. RT-PCR was used to determine the expressions of mRNA-PUMA, Cell viability and cell apoptosis were observed by MTT and flow cytometry （FCT）;PC-3 cells were transfected using siRNA-PUMA formulated with LipofectamineTM 2000.The expressions of mRNA-PUMA, cell viability and cell apoptosis was observed. Results： The expression of mRNA-PUMA in PC-3 cell line increased after the treatment of 5F.The proliferation of PC-3 cell line was greatly inhibited, and the apoptosis was greatly increased. Alter the PC-3 cell line was transfected into siRNA-PUMA.no apparent expression of mRNA-PUMA was observed after the treatment of 5F,and compared to controls, the proliferation and apoptosis rate did not change.Conclusions： 5F can up-regulate the expression of PUMA gene. which may inhibit apoptosis in pancreatic carcinoma, resulting in inhibition of tumor grow.