首页|CSTF3基因过表达对Nthy-ori 3-1细胞功能的影响

CSTF3基因过表达对Nthy-ori 3-1细胞功能的影响

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[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。
Effects of CSTF3 gene overexpression on Nthy-ori 3-1 cell function
[Objective]To study the function and mechanism of CSTF3 in thyroid cancer.[Method]CSTF3 overexpression Nt-hy-ori 3-1 cell line was constructed through lentiviral infection screening,CCK8 and cell cloning experiments were used to de-tect 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 OE cells were respectively:16.83%±4.61%,60.88%±9.26%(P<0.05);the number of clones formed in NC and OE cells at 7 days were:55.67±4.11,109.00±6.68(P<0.05);the 24 h apoptosis rates of NC and OE 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).[Conclu-sion]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 nu-cleotide mutations in chromosome structure and affect the expression of genes such as cycle and anti-apoptosis.

thyroid cancerCSTF3cell functioncell apoptosiscell proliferationnext-generation sequencingNthy-ori 3-1lentivirus

肖林、曹师瑜、谢依婷、龙芯仪、曹婷、叶丰、姜勇

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四川大学华西医院病理科,四川成都 610000

四川大学华西医院临床病理研究所,四川成都 610000

四川大学华西医院感染与疫苗研究室,四川成都 610000

甲状腺癌 CSTF3 细胞功能 细胞凋亡 细胞增殖 二代测序 Nthy-ori 3-1 慢病毒

国家自然科学基金面上项目四川省科技计划重点研发项目四川省自然科学基金项目

8217285023ZDYF246323NSFSC1623

2024

生物技术
黑龙江省微生物学会 黑龙江省生物工程学会 黑龙江省科学院微生物研究所

生物技术

CSTPCD
影响因子:0.611
ISSN:1004-311X
年,卷(期):2024.34(3)