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目的观察雷公藤内酯醇对多发性骨髓瘤细胞RPMI 8226增殖和周期的影响,探讨细胞周期调控蛋白P21wap1/cip1和P27kip1在其中的作用和意义。方法采用MTT比色法和流式细胞术检测雷公藤内酯醇对RPMI 8226细胞增殖、凋亡和细胞周期的影响,半定量RT-PCR和Western blotting法检测RPMI 8226细胞中P21wap1/cip1、P27kip1 mRNA和蛋白表达。结果 雷公藤内酯醇能明显抑制RPMI 8226细胞增殖,其抑制作用呈时间、剂量依赖性,雷公藤内酯醇作用48 h的IC50值为(71.18±2.01)nmol/L。雷公藤内酯醇还可以诱导RP-MI 8226细胞周期阻滞于G0/G1期,随着雷公藤内酯醇浓度的增加,G0/G1期细胞逐渐增多,S期细胞逐渐减少。经雷公藤内酯醇干预后周期调节蛋白P21wap1/cip1和P27kip1的mRNA和蛋白表达水平明显上调。结论雷公藤内酯醇可以抑制RPMI 8226细胞增殖,该抑制作用是通过调控P21wap1/cip1和P27kip1的表达,从而阻止细胞周期G0/G1期过渡实现的。
Objective To observe the effects of triptolide on the proliferation and cell cycle of multiple myeloma RPMI 8226 cells and to explore the role and significance of the cell cycle regulatory proteins P21wap1 / cip1 and P27kip1. Methods The effects of triptolide on the proliferation, apoptosis and cell cycle of RPMI 8226 cells were detected by MTT assay and flow cytometry. The expressions of P21wap1 / cip1 and P27kip1 mRNA in RPMI 8226 cells were detected by semi-quantitative RT-PCR and Western blotting And protein expression. Results Triptolide significantly inhibited the proliferation of RPMI 8226 cells in a time and dose dependent manner. The IC50 value of triptolide for 48 h was (71.18 ± 2.01) nmol / L. Triptolide also induced cell cycle arrest in G0 / G1 phase of RP-MI 8226 cells. With the increase of triptolide concentration, cells in G0 / G1 phase increased gradually and cells in S phase decreased gradually. The levels of mRNA and protein of P21wap1 / cip1 and P27kip1 were significantly up-regulated after triptolide intervention. Conclusion Triptolide can inhibit the proliferation of RPMI 8226 cells by inhibiting the expression of P21wap1 / cip1 and P27kip1, thereby preventing the cell cycle G0 / G1 phase transition.