Yang Yan, Zhao Feng
Department of Clinical Pharmacy, No. 1 People's Hospital, Jining, Shandong, China.
Department of Neurosurgery, Jining No. 1 People's Hospital, Jining, Shandong, China.
Arch Med Sci. 2020 May 25;20(3):839-846. doi: 10.5114/aoms.2020.95653. eCollection 2024.
Gliomas are lethal cancers accounting for significant human mortality across the globe. MicroRNAs (miRs) have shown potential to act as therapeutic targets for the treatment of cancer. Herein the role and therapeutic implications of miR-16 in glioma were investigated.
Expression analysis was carried out by qRT-PCR. Cell-Titer-Glo assay (Promega) was used for the determination of cell proliferation. DAPI, AO/EB, and annexin V/PI assays were used to detect apoptosis. Wound healing and Transwell assays were used for cell migration and invasion, respectively. Western blot analysis was used for the determination of protein expression.
The study revealed that miR-16 was significantly suppressed in the human glioma cells. Ectopic expression of miR-16 in U118 MG cells inhibited the proliferation via induction of apoptosis. The apoptosis induction was also accompanied by an upsurge of Bax and depletion of Bcl-2. The overexpression of miR-16 also inhibited the migration and invasion of the glioma U118 MG cells, as evident from the wound healing and transwell assays, which were accompanied by the inhibition of metalloproteinase-2 and -9 (MMP-2 and MMP-9). The effects of miR-16 overexpression were also examined on the PI3K/AKT/mTOR signalling pathway. The results showed that miR-16 overexpression inhibited the phosphorylation of the p70S6K, AKT, and mTOR at Ser, Ser, and Thr389, respectively, with no apparent effects on the total PI3K and AKT.
miR-16 acts as tumour suppressor in glioma and may severe as therapeutic target for glioma treatment.
胶质瘤是致命性癌症，在全球导致大量人类死亡。微小RNA（miR）已显示出作为癌症治疗靶点的潜力。本文研究了miR-16在胶质瘤中的作用及治疗意义。
通过qRT-PCR进行表达分析。使用Cell-Titer-Glo检测法（普洛麦格公司）测定细胞增殖。采用DAPI、AO/EB和膜联蛋白V/PI检测法检测细胞凋亡。分别使用伤口愈合实验和Transwell实验检测细胞迁移和侵袭。采用蛋白质印迹分析测定蛋白质表达。
研究表明，miR-16在人胶质瘤细胞中显著下调。在U118 MG细胞中异位表达miR-16可通过诱导凋亡抑制增殖。凋亡诱导还伴随着Bax的增加和Bcl-2的减少。如伤口愈合实验和Transwell实验所示，miR-16的过表达也抑制了胶质瘤U118 MG细胞的迁移和侵袭，同时伴随着金属蛋白酶-2和-9（MMP-2和MMP-9）的抑制。还研究了miR-16过表达对PI3K/AKT/mTOR信号通路的影响。结果表明，miR-16过表达分别抑制了p70S6K、AKT和mTOR在Ser、Ser和Thr389位点的磷酸化，对总PI3K和AKT没有明显影响。
miR-16在胶质瘤中起肿瘤抑制作用，可能作为胶质瘤治疗的靶点。
