Sehati Nasser, Sadeghie Navaz, Mansoori Behzad, Mohammadi Ali, Shanehbandi Dariush, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
J Cell Biochem. 2020 Jan;121(1):458-467. doi: 10.1002/jcb.29211. Epub 2019 Jun 25.
Melanoma skin cancer is one of the main causes of male cancer-related deaths worldwide. It has been suggested that miR-330-5p can act as a tumor suppressor in various types of cancers. So, in this study, we replaced miR-330 in melanoma cancer cells by vector-based miR-330 to evaluate the effects of this microRNA on the growth and migration inhibition of melanoma cancer cells, and to determine the molecular mechanisms underlying its action. By using the MTT assay, the IC of Geneticin antibiotic was obtained as 460 µg/mL. The results of the qRT-PCR showed the increased expression level of miR-330 and decreased expression levels of MMP-9, CXCR4, Vimentin, melanoma cell adhesion molecule, AKT1, and E2F1 messenger RNA in A375 transfected cells. The cytotoxicity assay results demonstrated the inhibition of cancer cells proliferation. Furthermore, the wound healing test results showed a migration reduction of transfected cells with miR-330 compared with nontransfected ones. In addition, 4',6-diamidino-2-phenylindoleLB: Luria-Bertani (DAPI) staining revealed the significant nucleus fragmentation in miR-330 replaced cells, which correspond to apoptosis induction in replaced cells. The results showed that increase in miR-330 expression level could significantly inhibit the tumor cell growth and the migration of melanoma cancer cells.
皮肤黑色素瘤是全球男性癌症相关死亡的主要原因之一。有人提出，miR-330-5p在多种类型的癌症中可作为一种肿瘤抑制因子。因此，在本研究中，我们通过基于载体的miR-330替代黑色素瘤癌细胞中的miR-330，以评估这种微小RNA对黑色素瘤癌细胞生长和迁移抑制的影响，并确定其作用的分子机制。通过MTT试验，获得遗传霉素抗生素的半数抑制浓度（IC）为460μg/mL。qRT-PCR结果显示，在A375转染细胞中，miR-330表达水平升高，而基质金属蛋白酶-9（MMP-9）、趋化因子受体4（CXCR4）、波形蛋白、黑色素瘤细胞粘附分子、蛋白激酶B1（AKT1）和E2F1信使核糖核酸的表达水平降低。细胞毒性试验结果表明癌细胞增殖受到抑制。此外，伤口愈合试验结果显示，与未转染细胞相比，miR-330转染细胞的迁移减少。另外，4',6-二脒基-2-苯基吲哚（DAPI）染色显示miR-330替代细胞中存在明显的细胞核碎片化，这与替代细胞中的凋亡诱导相对应。结果表明，miR-330表达水平的升高可显著抑制黑色素瘤癌细胞的肿瘤细胞生长和迁移。
