三叶青黄酮抑制肺癌A549细胞生长与蛋白酶体活性的关系研究

Radix tetrastigma hemsleyani flavone suppresses human lung carcinoma A549 cell via proteasome inhibition

目的:探讨三叶青黄酮对非小细胞肺癌A549细胞增殖抑制作用及其机制。方法:采用MTT法检测三叶青黄酮对A549细胞增殖的抑制作用;酶标仪检测细胞蛋白酶体和DUB活性;Real-time PCR和Western blot法检测细胞泛素-蛋白酶体途径相关蛋白ub-prs、USP14、UCHL5、POH1等表达变化。结果:体外实验MTT法检测三叶青黄酮对非小细胞肺癌A549细胞增殖的抑制作用,显示三叶青黄酮各浓度组的抑制率比较,差异具有统计学意义,并呈剂量依赖性。酶标仪检测结果显示随三叶青黄酮浓度升高,细胞蛋白酶体和DUB活性逐渐降低,呈明显的量-效关系。Real-time PCR和Western blot法检测表明三叶青黄酮可以上调A549细胞中ub-prs蛋白表达,下调USP14、UCHL5、POH1蛋白表达,呈剂量依赖性。结论:三叶青黄酮对非小细胞肺癌A549细胞具有明显抑制增殖作用,其机制可能与调节泛素-蛋白酶体途径有关。

AIM： To study the effect of radix tetrastigma hemsleyani flavone （RTHF） on the growth in human lung carcinoma A549 cells as well as its mechanisms. METHODS： A549 cells were treated with the RTHF in different concentrations. The proliferation of A549 cell was detected by MTI＂ assay in vitro. Enzyme proteasome assay was used to detect the activity of proteasome and DUB. Expressions of ubiquitin proteasome pathway associated proteins were detected by Real-time PCR and Western blot. RESULTS：MTr showed that RTHF had obvious anti-proliferation effect on A549 cells in a dosedependent manner. Enzyme proteasome assay showed that the treatment of activity of proteasome and DUB was significantly lower than the proportion in the control group （ P 〈 0.05 ）. Real-time PCR and Western blot showed the protein expression of ub-prs and was up-regulated significantly and that of USP14, UCHL5, POHI were downregulated in RTHF group when compared with the control group. CONCLUSION： Radix tetrastigma hemsleyani flavone may inhibit the proliferation of lung cancer A549 cells through the ubiquitin proteasome pathway.