Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Biomed Pharmacother. 2019 Mar;111:460-467. doi: 10.1016/j.biopha.2018.12.094. Epub 2018 Dec 27.
Lung cancer is the main cause of cancer death in males and females worldwide. Reduced expression of miR-145 has been reported in many types of cancers. In this study, we transfected miR-145 into lung cancer cells by vector-based miR-145, and investigated the effects of this intervention on growth and migration inhibition of cancer cells as well on the expression of targeted genes.
IC50 of Geneticin (G418) antibiotic was measured using MTT test in NSCLC cell lines. miR-145 was transfected into lung cancer cells by jetPEI. qRT-PCR was used to evaluate the transcript level of the miR-145 and expression for KRAS, MMP-9, vimentin, caspase-3, caspase-8 and caspase-9 genes in A549 cells. MTT assay was used to evaluate the proliferation inhibition of cancer cells. Wound healing assay was used to check the migration status of transfected lung cancer cells. The apoptosis induction was assessed by DAPI staining assay.
The MTT assay showed that the IC50 of Genticin was 494.1 μg/ml. The results of the qRT-PCR showed increased expression level of miR-145 and downregulation of KRAS, MMP-9, and vimentin expression in A549 transfected cells compared with the control group. The MTT assay results demonstrated inhibition of cancer cell proliferation after miR-145 replacement. Wound healing assay results revealed that migration was reduced upon miR-145 transfection. The transfected cell displayed increased apoptosis rate by inducing caspase-3 and caspase-9 mRNA expression.
The results of this study showed that increased miR-145 expression exerted a critical role in subsiding the growth, survival, and migration of lung cancer cell line.
肺癌是全球男性和女性癌症死亡的主要原因。许多类型的癌症中都报道了 miR-145 的表达降低。在这项研究中，我们通过基于载体的 miR-145 将 miR-145 转染到肺癌细胞中，并研究了这种干预对癌细胞生长和迁移抑制以及对靶基因表达的影响。
MTT 试验测定 NSCLC 细胞系中遗传霉素（G418）抗生素的 IC50。通过 jetPEI 将 miR-145 转染到肺癌细胞中。qRT-PCR 用于评估 miR-145 的转录水平以及 A549 细胞中 KRAS、MMP-9、波形蛋白、caspase-3、caspase-8 和 caspase-9 基因的表达。MTT 试验用于评估癌细胞的增殖抑制。划痕愈合试验用于检查转染肺癌细胞的迁移状态。通过 DAPI 染色试验评估细胞凋亡诱导。
MTT 试验表明 Genticin 的 IC50 为 494.1μg/ml。qRT-PCR 结果显示，与对照组相比，转染 miR-145 的 A549 细胞中 miR-145 的表达水平升高，KRAS、MMP-9 和波形蛋白的表达下调。MTT 试验结果表明，miR-145 替代后癌细胞增殖受到抑制。划痕愈合试验结果表明，miR-145 转染后迁移减少。通过诱导 caspase-3 和 caspase-9 mRNA 表达，转染细胞显示出更高的凋亡率。
本研究结果表明，miR-145 表达增加在缓解肺癌细胞系的生长、存活和迁移中发挥了关键作用。
