冬凌草甲素对SW1990细胞的增殖抑制作用

Antiproliferation effects of oridonin on human pancreatic carcinoma SW1990 cells

背景与目的:中医药治疗肿瘤不良反应较小且疗效尚佳,在胰腺癌防治方面有较大的潜力与优势,正日益受到国内外医学界的关注。本研究观察中草药冬凌草甲素对人胰腺癌SW1990细胞的增殖抑制作用,并探讨其可能机制。方法:以不同浓度(12.5、25和50μmol/L)的冬凌草甲素作用于体外培养的SW1990细胞,采用MTT法检测细胞生长抑制率,光学显微镜下观察药物对细胞形态的影响。DAPI法染色后,用荧光显微镜观察细胞核凋亡,用流式细胞仪检测细胞凋亡率,实时荧光定量PCR检测细胞凋亡前后端粒酶hTERT mRNA表达水平的变化。结果:冬凌草甲素对人胰腺癌SW1990细胞具有明显的增殖抑制作用,荧光显微镜见到典型的细胞凋亡形态学改变。冬凌草甲素12.5、25和50μmol/L给药组早期细胞凋亡的百分率显著高于未给药组[(2.17±0.57)%、(5.83±0.57)%、(4.43±0.78)%vs(0.30±0.17)%,P<0.05],晚期细胞凋亡和坏死细胞的百分率也显著高于未给药组[(5.10±0.98)%、(7.03±1.52)%、(10.13±0.70)%vs(2.63±0.06)%,P<0.05]。冬凌草甲素12.5、25和50μmol/L给药组端粒酶hTERT mRNA的表达水平显著高于未给药组(7.72±0.33、8.17±0.67、10.36±1.65 vs 5.94±0.17,P<0.05)。结论:冬凌草甲素可抑制胰腺癌SW1990细胞增殖,促进肿瘤细胞凋亡,其作用机制可能与药物引起端粒酶hTERT mRNA的表达水平均降低有关。

Background and purpose：Traditional Chinese medicine is known for its effectiveness and few adverse reactions.It has been receiving increasing attention from the medical profession due to its potential and advantages in treating pancreatic carcinoma.This experiment studied the antiproliferation effects of oridonin on human pancreatic carcinoma SW1990 cells and its mechanisms.Methods：SW1990 cells in a culture medium were treated with different concentrations of oridonin.The inhibitory rate of the cells was measured by MTT assay.The morphological changes were observed using a light microscope.The morphology of cell apoptosis was observed by DAPI stain and cell apoptotic rate was detected by flow cytometry（FCM）.Real time fluorescence quantitative polymerase chain reaction（RT-PCR） was used to detect hTERT mRNA expression both before and after apoptosis.Results：The growth of human pancreatic carcinoma SW1990 cells was significantly inhibited by oridonin.Apoptosis morphological changes including the condensation of chromatin and nuclear fragmentation were clearly observed using a DAPI stain.The early apoptotic rate of SW1990 cells treated with 12.5,25,50 μmol/L oridonin were significantly higher than that of the control group’s [（2.17±0.57）%,（5.83±0.57）%,（4.43±0.78）% vs（0.30±0.17）%,P0.05].The late apoptotic and cell necrosis rate was also significantly higher than that of the control group’s [（5.10±0.98）%,（7.03±1.52）%,（10.13±0.70）% vs（2.63±0.06）%,P0.05].△Ct of hTERT mRNA expression was obviously higher than the control group’s（7.72±0.33,8.17±0.67,10.36±1.65 vs 5.94±0.17,P0.05）.Conclusion：Oridonin can noticeably inhibit the proliferation and induce apoptosis of SW1990 cells.The mechanisms may be the down regulation of the hTERT mRNA expression after treatment with oridonin.