8-氯腺苷酸诱导多发性骨髓瘤细胞凋亡的实验研究

Apoptosis of Multiple Myeloma Cells Induced by 8-Cl-cAMP In Vitro

本研究探究8-氯腺苷酸对多发性骨髓瘤细胞的可能作用。以多发性骨髓瘤细胞株RPMI8226细胞为体外模型,在观察8-氯腺苷酸对细胞生长、活力及形态学影响的基础上,应用流式细胞术检测细胞DNA含量分布和细胞表面Annexin-V的含量,以DNA凝胶电泳技术评判细胞凋亡与否,并应用Western blot检测药物处理前后细胞内细胞周期调控蛋白cylinE和CDK2的变化。结果表明:低浓度8-氯腺苷酸(1-30μmol/L)能够明显地抑制RPMI8226细胞的生长,显著降低其细胞活力,且呈时间和浓度依赖性。进一步细胞形态学观察、流式细胞术及DNA凝胶电泳分析显示,这些浓度的8-氯腺苷酸诱导RPMI8226细胞凋亡。Western blot检测表明,8-氯腺苷酸可通过影响细胞周期调控蛋白cylin E和CDK2的表达而干扰细胞增殖。结论:8-氯腺苷酸能够有效地抑制多发性骨髓瘤细胞生长并诱导其凋亡。

The study was aimed to investigate the possible effects of 8-chloroadenosine 3＇, 5＇-monophosphate （ 8-C1- cAMP） on the multiple myeloma cells. The multiple myeloma cell line RPMI8226 was used as in vitro models. The effect on growth inhibition of RPMI8226 cells was evaluated by cell growth and viability curve, DNA fragment was visualized by agarose gel electrophoresis. The amount of apoptosis cells was measured by flow cytometry. Meanwhile Western blot assay were used to detect the change of several key cell cycle regulatory proteins CDK2 and cyclin E in these cells before and after the treatment. The results showed that low dose 8-CI-cAMP （1 -30 μmol/L） inhibited the proliferation and viability of RPMI8226 cells significantly. Agarose gel electrophoresis of DNA revealed the apoptosis characteristic “ladder”pattern. Apoptosis was also confirmed by flow cytometry, In addition, 8-CI-cAMP was able to inhibit the cell growth through modulating expression of cell cycle regulators CDK2 and cyclin E. It is concluded that 8-cl- cAMP inhibits the proliferation and induce apoptosis of multiple myeloma cells effectively.