LncRNA SNHG12靶向结合miR-206对甲状腺乳头状癌增殖、凋亡及AKT3蛋白的作用机制

The mechanism of lncRNA SNHG12 targeting miR-206 on proliferation and apoptosis of thyroid papillary carcinoma and AKT3 protein

目的研究lncRNA SNHG12靶向结合miR-206对甲状腺乳头状癌增殖、凋亡及AKT3蛋白的作用机制。方法选取2020年1月~2021年1月于新乡市中心医院接受治疗的甲状腺乳头状癌患者的癌组织与癌旁组织标本各10例作为研究对象。将甲状腺乳头状癌细胞系TPC-1细胞分成3组:实验组(TPC-1常规培养无处理组)、转染组(SNHG12基因沉默组)和对照组(转染不相关序列组)。RT-PCR检测lncRNA SNHG12、miR-206水平,CCK-8检测细胞增殖,流式细胞仪检测细胞凋亡,免疫印迹检测AKT3,双荧光素酶报告检测lncRNA SNHG12靶向miR-206,双荧光素酶报告检测向miR-206靶向AKT3。结果lncRNA SNHG12在甲状腺乳头状癌组织中的表达量高于癌旁组织(t=11.240,P<0.05),miR-206在甲状腺乳头状癌组织中的表达量低于癌旁组织(t=6.795,P<0.05)。lncRNA SNHG12在正常甲状腺细胞Nthy-ori 3-1中的表达水平最低,在TPC-1中的表达量高于在K1、BCPAP细胞中的表达(t=5.753,P<0.05)。miR-206在正常甲状腺细胞Nthy-ori 3-1中的表达水平最高,在TPC-1细胞中的表达低于在K1、BCPAP细胞中的表达(t=11.000,P<0.05)。与对照组相比,转染组lncRNA SNHG12表达水平、细胞凋亡率、AKT3蛋白表达降低(t=5.306,P<0.05;t=19.850,P<0.001;t=12.440,P<0.001),而miR-206表达升高(t=2.504,P<0.05),转染组在24、48、72 h的细胞增殖低于对照组(F=42.000、69.740、52.510,P<0.01)。结论低表达的lncRNA SNHG12通过靶向上调miR-206表达、抑制AKT3蛋白,从而抑制甲状腺乳头状癌细胞增殖并促进其凋亡。

OBJECTIVE To study the mechanism of lncRNA SNHG12 targeting and binding miR-206 on the proliferation,apoptosis and AKT3 protein of thyroid papillary carcinoma.METHODS From January 2020 to January 2021,10 patients with papillary thyroid carcinoma received treatment in Xinxiang Central Hospital were selected as the research objects.Thyroid papillary carcinoma cell line TPC-1 cells were divided into three groups:experimental group(TPC-1 routine culture group without treatment),transfection group(SNHG12 gene silencing group)and control group(transfection unrelated sequence group).lncRNA SNHG12 and miR-206 levels were detected by RT-PCR,cell proliferation was detected by CCK-8,cell apoptosis was detected by flow cytometry,AKT3 was detected by western blot,and lncRNA SNHG12 targeting miR-206was detected by dual luciferase reporting.Dual luciferase report assays toward miR-206 targeting AKT3.RESULTS The expression level of lncRNA SNHG12 in thyroid papillary carcinoma tissues was higher than that in para-carcinoma tissues(t=11.240,P<0.05),and the expression level of miR-206 in thyroid papillary carcinoma tissues was lower than that in para-carcinoma tissues(t=6.795,P<0.05).lncRNA SNHG12 had the lowest expression level in normal thyroid cells THY ori 3-1,and the expression level in TPC-1 was higher than that in K1 and BCPAP cells(t=5.753,P<0.05).The expression level of miR-206was the highest in normal thyroid cells Nthy-ori 3-1,and the expression level in TPC-1 cells was lower than that in K1 and BCPAP cells(t=11.000,P<0.05).Compared with control group,lncRNA SNHG12 expression level,apoptosis rate and AKT3protein expression in transfection group were decreased(t=5.306,P<0.05;t=19.850,P<0.001;t=12.440,P<0.001),while miR-206 expression was increased(t=2.504,P<0.05),and cell proliferation in the transfection group was lower than that in the control group at 24,48 and 72 h(F=42.000,69.740,52.510,P<0.01).CONCLUSION lncRNA SNHG12 with low expression can inhibit the proliferat.