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[慢病毒介导的LAMP2A基因短发夹RNA沉默抑制多发性骨髓瘤细胞增殖]

[Lentivirus-mediated shRNA silencing of LAMP2A inhibits the proliferation of multiple myeloma cells].

作者信息

Li Lixuan, Li Jia

机构信息

Interdisciplinary Higher Institute of Science and Technology, East China Normal University, Shanghai 200062; State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, SIBS, Chinese Academy of Sciences, Shanghai 201203, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2015 May;31(5):605-8, 614.

Abstract

OBJECTIVE

To study the effects of lentivirus-mediated short hairpin RNA (shRNA) silencing of lysosome-associated membrane protein type 2A (LAMP2A) expression on the proliferation of multiple myeloma cells.

METHODS

The constructed shRNA lentiviral vector was applied to infect human multiple myeloma cell line MM.1S, and stable expression cell line was obtained by puromycin screening. Western blotting was used to verify the inhibitory effect on LAMP2A protein expression. MTT assay was conducted to detect the effect of knocked-down LAMP2A on MM.1S cell proliferation, and the anti-tumor potency of suberoylanilide hydroxamic acid (SAHA) against the obtained MM.1S LAMP2A(shRNA) stable cell line. Lactate assay was performed to observe the impact of low LAMP2A expression on cell glycolysis.

RESULTS

The stable cell line with low LAMP2A expression were obtained with the constructed human LAMP2A-shRNA lentiviral vector. Down-regulation of LAMP2A expression significantly inhibited MM.1S cell proliferation and enhanced the anti-tumor activity of SAHA. Interestingly, decreased LAMP2A expression also inhibited MM.1S cell lactic acid secretion.

CONCLUSION

Down-regulation of LAMP2A expression could inhibit cell proliferation in multiple myeloma cells.

摘要

目的

研究慢病毒介导的溶酶体相关膜蛋白2A(LAMP2A)表达的短发夹RNA(shRNA)沉默对多发性骨髓瘤细胞增殖的影响。

方法

应用构建的shRNA慢病毒载体感染人多发性骨髓瘤细胞系MM.1S,经嘌呤霉素筛选获得稳定表达细胞系。采用蛋白质免疫印迹法验证对LAMP2A蛋白表达的抑制作用。进行MTT试验以检测敲低LAMP2A对MM.1S细胞增殖的影响,以及辛二酰苯胺异羟肟酸(SAHA)对所获得的MM.1S LAMP2A(shRNA)稳定细胞系的抗肿瘤效力。进行乳酸测定以观察低LAMP2A表达对细胞糖酵解的影响。

结果

用构建的人LAMP2A-shRNA慢病毒载体获得了LAMP2A表达低的稳定细胞系。LAMP2A表达下调显著抑制MM.1S细胞增殖并增强SAHA的抗肿瘤活性。有趣的是,LAMP2A表达降低也抑制了MM.1S细胞乳酸分泌。

结论

LAMP2A表达下调可抑制多发性骨髓瘤细胞的增殖。

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