MiR-103a-3p靶向FLOT2调控甲状腺癌细胞增殖、凋亡的分子机制
| 作者: |
1刘兵雄,
1李学刚
1 汉川市人民医院普外三科，湖北 汉川 431600 |
| 通讯: |
李学刚
Email: 355113715@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2020.07.006 |
摘要
目的：研究miR-103a-3p对甲状腺癌细胞增殖、凋亡的影响及其机制。方法：运用RT-qPCR法检测甲状腺癌细胞SW579，FTC-133及人甲状腺上皮细胞TEC中miR-103a-3p、脂筏结构蛋白2(FLOT2)的表达。将miR-NC(miR-NC组)、miR-103a-3p mimics(miR-103a-3p组)、anti-miR-NC(anti-miR-NC组)、anti-miR-103a-3p(anti-miR-103a-3p组)、si-NC(si-NC组)、si-FLOT2(si-FLOT2组)、miR-103a-3p+pcDNA mimics(miR-103a-3p+pcDNA组)、miR-103a-3p mimics+pcDNA-FLOT2(miR-103a-3p+pcDNA-FLOT2组)用脂质体法转染至SW579细胞。蛋白质印迹法检测细胞中FLOT2、多肿瘤抑制基因(P16)、细胞周期依赖性蛋白激酶抑制因子1A(P21)、聚腺苷二磷酸-核糖聚合酶[poly-(ADP-ribose)polymerase，PARP]、裂解的PARP(cleaved-PARP)、半胱天冬酶-3的前体(pro-caspase-3)、裂解的半胱天冬酶3(cleaved-caspase-3)的蛋白表达；细胞计数试剂盒(CCK-8)法检测细胞增殖；流式细胞术检测细胞凋亡；双荧光素酶报告基因检测实验检测细胞的荧光活性。结果：与人甲状腺上皮细胞TEC相比，甲状腺癌细胞中miR-103a-3p的表达明显下调，FLOT2的表达明显上调(P<0.05)；过表达miR-103a-3p、敲减FLOT2均可抑制甲状腺癌细胞的增殖，促进凋亡；miR-103a-3p明显抑制野生型FLOT2细胞的荧光活性，且负向调控FLOT2的表达；过表达FLOT2逆转miR-103a-3p对甲状腺癌细胞的增殖抑制和凋亡促进作用。结论：miR-103a-3p可抑制甲状腺癌细胞的增殖，促进凋亡，其机制可能与靶向FLOT2相关，可为甲状腺癌的治疗提供参考。
关键词：
miR-103a-3p；脂筏结构蛋白2；甲状腺癌
Molecular mechanism of miR-103a-3p targeting FLOT2 to regulate proliferation and apoptosis of thyroid cancer cells
CorrespondingAuthor: LI Xuegang Email: 355113715@qq.com
DOI: 10.3978/j.issn.2095-6959.2020.07.006
Abstract
Objective: To study the effect of miR-103a-3p on proliferation and apoptosis of thyroid cancer cells and its mechanism. Methods: RT-qPCR was used to detect the expression of miR-103a-3p and FLOT2 in thyroid cancer cells SW579, FTC-133 and human thyroid epithelial cells. MiR-NC group (transfected miR-NC), miR-103a-3p group (transfected miR-103a-3p mimics), anti-miR-NC (anti-miR-NC group), anti-miR-103a-3p (anti-miR-103a-3p group), si-NC (si-NC group), si-FLOT2 (si-FLOT2 group), miR-103a-3p mimics+pcDNA (miR-103a-3p + pc DNA group), miR-103a-3p mimics + pcDNA-FLOT2 (miR-103a-3p + pc DNA-FLOT2 group) were transfected with liposome method to SW579 cells, respectively. Western blot was used to detect the expression of FLOT2, P16, P21, cleaved-PARP and cleaved-caspase-3 in cells; CCK-8 method was used to detect cell proliferation; flow cytometry was used to detect apoptosis; dual luciferase reporter gene detection assay was used to detect the fluorescence activity of cells. Results: Compared with human thyroid epithelial TEC, the expression of miR-103a-3p was down-regulated and the expression of FLOT2 was up-regulated (P<0.05). Overexpression miR-103a-3p or knockdown FLOT2 inhibited proliferation and promoted apoptosis of thyroid cancer cells; miR-103a-3p significantly inhibits the fluorescence activity of wild-type FLOT2 cells and negatively regulates the expression of FLOT2; overexpression of FLOT2 reverses the proliferation inhibition of thyroid cancer cells by miR-103a-3p Apoptosis promoting effect. Conclusion: MiR-103a-3p can inhibit the proliferation and promote apoptosis of thyroid cancer cells. The mechanism may be related to the targeting of FLOT2, which will provide a reference for the treatment of thyroid cancer.
Keywords：
miR-103a-3p; FLOT2; thyroid cancer