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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。
Objective To investigate the effect of short hairpin RNA (shRNA) lentiviral vector on the proliferation of multiple myeloma cells by down - regulating the expression of lysosome - associated membrane protein 2A (LAMP2A) in multiple myeloma cells. Methods The constructed shRNA lentivirus vector was used to infect human MM.1S multiple myeloma cells. Puromycin was used to screen the stable cell lines. The inhibitory effect of LAMP2A protein was confirmed by Western blot. MTT assay was used to observe the effect of LAMP2A on the proliferation of MM.1S cells Effect and its effect on the anti-MM.1S cells of suberoylanilide hydroxamic acid (SAHA). The effect of LAMP2A on glycolysis was observed by lactate test. Results Multiple myeloma cell lines stably expressing LAMP2A were obtained based on the recombinant lentiviral vector of human LAMP2A-shRNA. The down-regulation of LAMP2A expression significantly inhibited the proliferation of MM.1S cells and enhanced the anti-tumor effect of SAHA against myeloma cells LAMP2A expression inhibits lactate secretion in MM.1S cells. Conclusion The down-regulation of LAMP2A expression in multiple myeloma cells can inhibit the proliferation of multiple myeloma cells.