熊果酸对肺癌细胞株A549及SPCA1细胞周期的抑制作用

目的 研究熊果酸(ursolic acid,UA)对肺癌细胞株A549及SPCA1细胞周期的抑制作用,探讨其分子机制。方法 采用MTS法检测不同浓度UA作用不同时间对A549及SPCA1细胞的增殖抑制作用;光镜下观察细胞形态学变化;流式细胞仪检测不同浓度UA对A549及SPCA1细胞周期分布的影响;实时荧光定量PCR分析UA作用下A549及SPCA1的相关分子表达情况。结果 MTS结果显示UA对A549及SPCA1细胞具有增殖抑制作用,且呈时间和剂量依赖性。经UA作用后,光镜下可见A549及SPCA1细胞增殖受到抑制,细胞出现变圆、回缩和脱落。流式细胞仪结果显示在UA作用后,A549及SPCA1细胞周期抑制在G₀/G₁期,S期和G₂/M期的比例降低。实时荧光定量PCR结果表明,UA作用后A549及SPCA1细胞cyclinD1 mRNA、cyclinE mRNA、Ankrd17 mRNA表达水平降低,p27 mRNA、p16 mRNA表达增加,Ankrd17 mRNA表达变化差异无统计学意义。结论 UA能明显抑制A549及SPCA1细胞增殖,呈时间和剂量依赖性,该作用是通过将细胞周期抑制在G₀/G₁期实现的。其分子机制可能与上调p27和p16的表达,下调cyclinD1和cyclinE的表达有关。

Effects of Ursolic Acid on Cell Cycle of A549 Cells and SPCA1 Cells and Its Mechanism

Objective To explore the cell cycle inhibition of ursolic acid in the lung cancer cell line A549 and and SPCA1 and stndy its mechanisms. Methods The A549 cells and SPCA1 cells were treated with ursolic acid at different concentration for different duration.The cell inhibition rate was measured by MTS assay. The morphology of A549 cells and SPCA1 cells were observed under inverted microscope.The flow cytometry was used to detect cell cycle arrest. The expression of cell cycle arrest related genes were detected with real-time pcr. Results Ursolic Acid remarkably inhibited the proliferation of A549 cells and SPCA1 cells and there was a dose and time-dependent manner among them.The morphological changes of A549 cells and SPCA1 cells, such as proliferation inhibition,cell retraction,exfoliation and necrosis were obvious under light microscope.The cell cycle was blocked at G₀/G₁ phase and the cell population in S and G₂/M phase was decreased.Real-time PCR analysis indicated that expression level of cyclinD1 mRNA, cyclinE mRNA, Ankrd17 mRNA was decreased while expression level of P27 mRNA, P16 mRNA was increased. The differential expression of Ankrd17 mRNA was not statistically significant. Conclusion Ursolic acid can inhibit the proliferation of A549 cells and SPCA1 cells and there is a dose and time-dependent manner.The effect maybe related with cell cycle arrest, which maybe associated with downregulating the expression level of cyclinD1, cyclinE and upregulating the expression level of p27, p16.