白桦脂酸联合硼替佐米诱导U266细胞凋亡及其机制研究Research on Betulinic Acid Combined with Bortezomib Inducing U266 Cell Apoptosis and Its mechanism
孙嘉,马丹,王萍,方琴
SUN Jia,MA Dan,WANG Ping,FANG QIN
摘要(Abstract)：
目的:探讨白桦脂酸(BA)联合硼替佐米诱导多发性骨髓瘤(MM)U266细胞凋亡及其机制。方法:用不同浓度的BA(20、40、60及80 mg/L)、硼替佐米(10、80及320 nmol/L)、40 mg/L BA分别联合10、80及320 nmol/L硼替佐米(联合1~3组)处理U266细胞,MTT法检测上述各组U266细胞增殖抑制率;Real-time PCR和蛋白免疫印迹法(Western blot)分别检测40 mg/L BA、80 nmol/L硼替佐米及40 mg/L BA联合80 nmol/L硼替佐米(联合4组)的U266细胞Survivin、Cyto-C、BCL-2及Bax的mRNA和蛋白表达水平。结果:联合1~3组对U266细胞增殖抑制率显著高于BA组和硼替佐米组(P<0.05);与硼替佐米组和BA组相比,联合4组U266细胞的cyto-C,Bax mRNA和蛋白表达水平显著升高(P<0.05),而Survivin、Bcl-2 mRNA和蛋白表达水平显著降低(P<0.05)。结论:BA联合硼替佐米可以促进MM肿瘤细胞凋亡,机制可能与下调Survivin、Bcl-2表达,上调Cyto-c、Bax表达有关。
Objective: To investigate betulinic acid combined with bortezomib inducing apoptosis of multiple myeloma( MM) U266 cells and its mechanism. Method: U266 cells were treated with betulinic acid( 20 mg / L,40 mg / L,60 mg / L,80 mg / L), bortezomib( 10 nmol / L,80 nmol / L,320 nmol / L),BA( 40 mg / L) combined with 10 nmol / L,80 nmol / L,320 nmol / L bortezomib( combination 1 ~ 3 groups). The proliferation inhibition rate of U266 cells was detected by MTT assay; Real-time PCR and Western blot were applied to detect mRNA and protein expression levels of Survivin,Cyto C,of BCL-2 and Bax in 40 mg / L BA group,80 nmol / L bortezomib group and 40 mg / L BA combined with 80 nmol / L bortezomib group. Results: Proliferation inhibition rate of U266 cells in combination 1 ~ 3 groups was significantly higher than that in BA group and bortezomib group( P < 0. 05);compared with BA group and bortezomib group,mRNA and protein expression levels of Cyto C,Bax in the combination 4 group were remarkably increased( P < 0. 05),the Survivin,Bcl-2 mRNA and protein expression levels were significantly decreased( P < 0. 05). Conclusions: The betulinic acid combined with bortezomib is proved to induce apoptosis of MM cells and its mechanism may be associated with the down-regulation of Survivin,BCL-2 and up-regulation of Cyto-c,Bax expression levels.
关键词(KeyWords)：
白桦脂酸;硼替佐米;U266细胞;凋亡;基因表达
betulinic acid;borezomib;U266 cells;apoptosis;gene expression
基金项目(Foundation): 国家自然科学基金(No:81360501)
作者(Author):
孙嘉,马丹,王萍,方琴
SUN Jia,MA Dan,WANG Ping,FANG QIN
DOI: 10.19367/j.cnki.1000-2707.2015.10.005
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