Rodrigues-Junior Dorival Mendes, Pelarin Maria Fernanda de Andrade, Nader Helena Bonciani, Vettore André Luiz, Pinhal Maria Aparecida Silva
Department of Biochemistry, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.
Institute of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden.
Onco Targets Ther. 2021 Jan 15;14:455-467. doi: 10.2147/OTT.S286751. eCollection 2021.
Multiple myeloma (MM) remains an incurable disease, and patient survival requires a better understanding of this malignancy's molecular aspects. Heparanase (HPSE) is highly expressed in aggressive MM cells and related to tumor growth, metastasis, and bortezomib (BTZ) resistance. Thus, targeting HPSE seems to be a promising approach for MM treatment, and because microRNAs (miRNAs) have emerged as potential regulators of HPSE expression, the use of extracellular vesicles (EVs) can allow the efficient delivery of therapeutic miRNAs.
We used prediction algorithms to identify potential miRNAs that regulate negatively HPSE expression. RT-qPCR was performed to assess miRNAs and HPSE expression in MM lines (U266 and RPMI-8226). Synthetic miRNA mimics were electroporated in MM cells to understand the miRNA contribution in HPSE expression, glycosaminoglycans (GAGs) profile, cell proliferation, and cell death induced by BTZ. EVs derived from HEK293T cells were engineered with miRNAs to evaluate their therapeutic potential combined with BTZ.
It revealed a direct association between BTZ sensitivity, HPSE, and miR-1252-5p expressions. Moreover, overexpression of miR-1252-5p significantly reduced HPSE expression and HPSE enzymatic activity in MM cells. The higher level of miR-1252-5p was correlated with a reduction of cell viability and higher sensitivity to BTZ. Further, EVs carrying miR-1252-5p increased MM cells' sensitivity to BTZ treatment.
These results showed that miR-1252-5p could negatively regulate HPSE in MM, indicating the use of EVs carrying miR-1252-5p as a potential novel BTZ sensitization approach in MM cells.
多发性骨髓瘤（MM）仍然是一种无法治愈的疾病，患者的生存需要对这种恶性肿瘤的分子层面有更好的了解。乙酰肝素酶（HPSE）在侵袭性MM细胞中高表达，与肿瘤生长、转移和硼替佐米（BTZ）耐药相关。因此，靶向HPSE似乎是MM治疗的一种有前景的方法，并且由于微小RNA（miRNA）已成为HPSE表达的潜在调节因子，细胞外囊泡（EV）的使用可以实现治疗性miRNA的有效递送。
我们使用预测算法来识别负向调节HPSE表达的潜在miRNA。进行逆转录定量聚合酶链反应（RT-qPCR）以评估MM细胞系（U266和RPMI-8226）中的miRNA和HPSE表达。将合成的miRNA模拟物电穿孔导入MM细胞，以了解miRNA在HPSE表达、糖胺聚糖（GAG）谱、细胞增殖以及BTZ诱导的细胞死亡中的作用。用miRNA对源自人胚肾293T细胞（HEK293T）的EV进行工程改造，以评估其与BTZ联合使用的治疗潜力。
研究揭示了BTZ敏感性、HPSE和miR-1252-5p表达之间的直接关联。此外，miR-1252-5p的过表达显著降低了MM细胞中HPSE的表达和HPSE酶活性。miR-1252-5p水平的升高与细胞活力降低以及对BTZ更高的敏感性相关。此外，携带miR-1252-5p的EV增加了MM细胞对BTZ治疗的敏感性。
这些结果表明，miR-1252-5p可以负向调节MM中的HPSE，这表明使用携带miR-1252-5p的EV作为MM细胞中一种潜在的新型BTZ增敏方法。
