Lu Zhongwu, Wu Zhiqiang, Hu Jiaqia, Wei Wenjun, Ma Ben, Wen Duo
Department of Head and Neck Surgery, Fudan University, Shanghai Cancer Center, Shanghai 200032, China.
J BUON. 2019 Sep-Oct;24(5):2114-2119.
Thyroid cancer causes significant mortality and 1-1.5% of all the new diagnosed cancers are thyroid cancers. The incidence of thyroid cancer is increasing at an alarming rate and therapeutic targets are lacking. This study was undertaken to investigate the role and therapeutic implications of microRNA (miR)-15 in thyroid cancer.
Expression analysis was performed by qRT-PCR. Transfections were performed by Lipofectamine 2000 reagent. The cell viability was determined by MTT assay. Apoptosis was detected by acridine orange (AO)/ethidium bromide (EB). The percentage of apoptotic cells was estimated by annexin V/ propidium iodide (PI) staining. Wound healing and transwell assays were used to monitor the cell migration and invasion. Protein expression was determined by western blotting.
The expression of miR-15 was found significantly decreased in thyroid cancer cells. Ectopic expression of miR-15 promoted the apoptosis of MDA-T35 thyroid cancer cells. The percentage of apoptotic MDA-T35 cells was 1.9% in thyroid cancer and 40.1% in miR-15 mimics transfected cells. The apoptosis promoted by miR-15 overexpression was also associated with enhancement of Bax and depletion of Bcl-2 in MDA-T35 cells. The TargetScan analysis showed Bcl-2 to be the target of miR-15. Additionally, the expression of Bcl-2 was also enhanced in all the thyroid cancer cells and miR-15 ectopic expression could cause suppression of the Bcl-2 expression in MDA-T35 cells. The wound healing assay showed that miR-5 overexpression caused decrease in the migration of MDA-T35 cells while the transwell assays showed decline in the invasion of miR-15 mimics transfected MDA-T35 cells.
To sum up, miR-15 may exhibit therapeutic implications in thyroid cancer and may prove useful in thyroid cancer treatment.
甲状腺癌导致显著的死亡率，所有新诊断癌症中有1 - 1.5%为甲状腺癌。甲状腺癌的发病率正以惊人的速度上升，且缺乏治疗靶点。本研究旨在探讨微小RNA（miR）-15在甲状腺癌中的作用及治疗意义。
通过qRT-PCR进行表达分析。使用Lipofectamine 2000试剂进行转染。通过MTT法测定细胞活力。用吖啶橙（AO）/溴化乙锭（EB）检测细胞凋亡。通过膜联蛋白V/碘化丙啶（PI）染色估计凋亡细胞的百分比。采用伤口愈合实验和Transwell实验监测细胞迁移和侵袭。通过蛋白质印迹法测定蛋白质表达。
发现miR-15在甲状腺癌细胞中的表达显著降低。miR-15的异位表达促进了MDA-T35甲状腺癌细胞的凋亡。甲状腺癌中凋亡的MDA-T35细胞百分比为1.9%，而在转染miR-15模拟物的细胞中为40.1%。miR-15过表达促进的凋亡也与MDA-T35细胞中Bax的增加和Bcl-2的减少有关。TargetScan分析显示Bcl-2是miR-15的靶标。此外，所有甲状腺癌细胞中Bcl-2的表达也增强，miR-15的异位表达可导致MDA-T35细胞中Bcl-2表达的抑制。伤口愈合实验表明，miR-15过表达导致MDA-T35细胞迁移减少，而Transwell实验显示转染miR-15模拟物的MDA-T35细胞侵袭能力下降。
综上所述，miR-15可能在甲状腺癌中具有治疗意义，可能对甲状腺癌治疗有用。
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