摘要
目的:探讨苦瓜蛋白MAP30在多发性骨髓瘤中的作用及其机制。方法:将人骨髓瘤RPMI-8226、NCI-H929和U266细胞为靶细胞,以不同浓度(1～10μmol/L)的MAP30处理后,采用CCK-8法检测多发骨髓瘤细胞株RPMI-8226、NCI-H929和U266的增殖能力,Annexin V/PI流式细胞术检测骨髓瘤细胞的凋亡情况,Wb实验检测骨髓瘤细胞凋亡相关蛋白(PARP)、自噬相关蛋白(LC3II、P62)及AKT/mTOR通路相关蛋白的表达水平,CCK-8法、流式细胞术和Wb实验检测MAP30联合自噬激动剂雷帕霉素(Rap)或自噬抑制剂巴弗洛霉素(Baf)作用后骨髓瘤细胞增殖、凋亡和自噬的变化。结果:MAP30(1～10μmol/L)抑制骨髓瘤细胞的增殖,且其对增殖的抑制呈时间和剂量依赖关系(P<0.05或P<0.01)。MAP30单独作用骨髓瘤细胞后,细胞凋亡和细胞自噬增加,PARP裂解增多,LC3II表达水平增加,P62表达明显下降(均P<0.05或P<0.01);MAP30+Rap作用骨髓瘤细胞后,较单用MAP30细胞增殖率增加、细胞凋亡率减少、细胞自噬减少、PARP裂解降低、LC3II表达降低及P62表达增加(均P<0.05或P<0.01);相反,MAP30+Baf作用骨髓瘤细胞较单用MAP30,细胞增殖率减少、细胞凋亡率增加、细胞自噬增加、PARP裂解增加、LC3II表达增加、P62表达降低(均P<0.05或P<0.01);MAP30作用骨髓瘤细胞后,AKT/mTOR通路相关蛋白p-AKT和p-mTOR表达水平明显降低(均P<0.05)。结论:MAP30可通过AKT/mTOR通路促进骨髓瘤细胞的凋亡和自噬,可能为骨髓瘤的治疗提供新的治疗策略。
Objective:To investigate the role of momordica protein MAP30 in multiple myeloma(MM)and the possible mechanism.Methods:Human myeloma RPMI-8226,NCI-H929 and U266 cells were treated with MAP30 at different concentration(1-10μmol/L)and then the proliferation rates of cells were detected by CCK-8 assay.Annexin V/PI flow cytometry was used to evaluate the apoptosis rate of myeloma cells,and the expressions of apoptosis-related protein(PARP),autophagy-related proteins(LC3II,P62)and Akt/mTOR pathway-related proteins in multiple myeloma cells were also detected via Wb.The changes in cell proliferation,apoptosis and autophagy after the treatment of MAP30 combined with autophagy agonist rapamycin(Rap)or autophagy inhibitor bafilomycin(Baf)were observed by CCK-8,flow cytometry and Wb,respectively.Results:MAP30(1-10μmol/L)inhibited the proliferation of myeloma cells in a time-and dose-dependent manner(P<0.05 or P<0.01).With MAP30 acting on myeloma cells alone,the apoptosis and autophagy of MM cells,as well as the expression of PARP cleavage and LC3II increased while the expression of P62 decreased significantly(all P<0.05 or P<0.01).After being treated with MAP30+Baf,compared with MAP30 treatment alone,the cell proliferation was remarkably enhanced while cell apoptosis and cell autophagy were suppressed,besides,the expression of PARP cleavage and LC3II were decreased and P62 level was augmented(all P<0.05 or P<0.01).Conversely,after being treated with MAP30+Baf,compared with MAP30 treat-ment or Baf treatment alone,cell proliferation and P62 level were reduced,while apoptosis and autophagy as well as the expressions of PARP cleavage and LC3II level were increased(all P<0.05 or P<0.01).The expressions of p-AKT and p-mTOR were significantly reduced with the effect of MAP30 on myeloma cells(all P<0.05).Conclusion:MAP30 can promote the apoptosis and autophagy of myeloma cells through AKT/mTOR pathway,which may provide a new therapeutic strategy for treatment of multiple myeloma.
作者
何颖
方芳
颜宏利
江松福
HE Ying;FANG Fang;YAN Hongli;JIANG Songfu(Department of Hematology,the First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325015,Zhejiang,China;Department of Laboratory Diagnosis,Changhai Hospital Affiliated to Naval Military Medical University,Shanghai 200433,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第3期299-305,共7页
Chinese Journal of Cancer Biotherapy
基金
浙江省自然科学基金资助项目(No.LY16H080006)~~
作者简介
何颖(1990-),女,硕士生,主要从事血液恶性肿瘤的研究,E-mail:490492763@qq.com;通信作者:江松福(JIANG Songfu,corresponding author),硕士,副主任医师,硕士生导师,主要从事血液恶性肿瘤的研究,E-mail:jiangsongfii@189.cn.
