CSTF3基因过表达对Nthy-ori 3-1细胞功能的影响

Effects of CSTF39gene overexpression on Nthy-ori 3-1 cell function

[目的]研究CSTF3在甲状腺癌发生中的功能及其机制。[方法]慢病毒感染筛选构建CSTF3过表达Nthy-ori 3-1细胞株,CCK8和细胞成克隆实验检测细胞增殖能力,划痕实验评估细胞迁移能力,流式细胞术检测细胞凋亡情况,紫外照射处理细胞后全外显子组测序,qPCR检测未照射及照射细胞周期、抗凋亡等基因表达。[结果]空载对照组(NC)、CSTF3过表达细胞株(OE)细胞72 h增殖率分别为:126.92%±19.57%、271.51%±27.21%(P<0.05);NC、OE细胞48 h迁移率分别为:16.83%±4.61%、60.88%±9.26%(P<0.05);NC、OE细胞7 d克隆形成个数分别为:55.67±4.11、109.00±6.68(P<0.05);NC、OE细胞24 h凋亡率分别为9.24%±0.47%、6.61%±0.08%(P<0.05)。CSTF3过表达细胞株染色体结构变异和单核苷酸突变数明显多于NC组。CSTF3过表达细胞株周期、抗凋亡等基因的表达水平显著高于NC组(P<0.05)。[结论]成功构建了CSTF3过表达的Nthy-ori 3-1细胞株,过表达促进Nthy-ori 3-1细胞增殖、迁移及成克隆能力,但抗细胞凋亡。提示CSTF3可能与染色体及单核苷酸突变相关,并且影响了周期和抗凋亡等基因的表达。

[Objective]To study the function and mechanism of CSTF3 in thyroid cancer.[Method]CSTF3 overexpression Nthy-ori 3-1 cell line was constructed through lentiviral infection screening,CCK8 and cell cloning experiments were used to detect cell proliferation ability,scratch test was used to evaluate cell migration ability,and flow cytometry was used to detect cell apoptosis.After the cells were treated with UV irradiation,the whole exome was sequenced,and qPCR was used to detect the expression of non-irradiated and irradiated cell cycle,anti-apoptosis and other genes.[Result]The 72 h cell proliferation rates of the empty control group(NC)and CSTF3 overexpression cell line(OE)were:126.92%±19.57%,271.51%±27.21%(P<0.05);the 48 h migration rates of NC and 0E cells were respectively:16.83%±4.61%,60.88%±9.26%(P<0.05);the number of clones formed in NC and 0E cells at 7 days were:55.67±4.11,109.00±6.68(P<0.05);the 24 h apoptosis rates of NC and 0E cells were respectively were 9.24%±0.47%and 6.61±0.08%(P<0.05).The results of cell whole exome sequencing showed that the number of chromosome structural variations and single nucleotide mutations in the CSTF3 overexpressing cell line was significantly more than that in the NC group.The expression levels of cycle and anti-apoptosis genes in CSTF3 overexpressing cell lines were significantly higher than those in the NC group(P<0.05).[Conclusion] An Nthy-ori 3-1 cell line overexpressing CSTF3 was successfully constructed.Overexpression promotes the proliferation,migration and cloning ability of Nthy-ori 3-1 cells,but resists apoptosis.It is suggested that CSTF3 may be related to single nucleotide mutations in chromosome structure and affect the expression of genes such as cycle and anti-apoptosis.