【摘要】 目的研究短发夹RNA(shRNA)慢病毒载体下调多发性骨髓瘤细胞中溶酶体相关膜蛋白2A(LAMP2A)表达对多发性骨髓瘤细胞增殖的影响。方法将构建的shRNA慢病毒表达载体感染人MM.1S多发性骨髓瘤细胞,嘌呤霉素筛选获得稳定细胞株,Western blot法验证LAMP2A蛋白表达抑制效果;MTT法观察LAMP2A对MM.1S细胞增殖的影响及其对辛二酰苯胺异羟肟酸(SAHA)抗MM.1S细胞的影响,乳酸检测观察LAMP2A对糖酵解的影响。结果在构建人LAMP2A-shRNA重组慢病毒载体的基础上,获得稳定低表达LAMP2A的多发性骨髓瘤细胞株;下调LAMP2A表达明显抑制MM.1S细胞的增殖并增强SAHA抗骨髓瘤细胞的作用,下调LAMP2A表达抑制MM.1S细胞乳酸分泌。结论下调多发性骨髓瘤细胞LAMP2A的表达可抑制多发性骨髓瘤细胞增殖。更多还原

【Abstract】 Objective To study the effects of lentivirus-mediated short hairpin RNA( shRNA) silencing of lysosomeassociated 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 LAMP2 A protein expression. MTT assay was conducted to detect the effect of knocked-down LAMP2 A on MM. 1S cell proliferation,and the anti-tumor potency of suberoylanilide hydroxamic acid( SAHA) against the obtained MM. 1S LAMP2 AshRNAstable cell line.Lactate assay was performed to observe the impact of low LAMP2 A expression on cel glycolysis. Results The stable cel line with low LAMP2 A expression were obtained with the constructed human LAMP2A-shRNA lentiviral vector. Down-regulation of LAMP2 A expression significantly inhibited MM. 1S cell proliferation and enhanced the anti-tumor activity of SAHA. Interestingly,decreased LAMP2 A expression also inhibited MM. 1S cell lactic acid secretion. Conclusion Down-regulation of LAMP2 A expression could inhibit cell proliferation in multiple myeloma cells.更多还原