文章摘要

过表达PTEN基因对甲状腺髓样癌细胞生长及细胞周期的影响

作者: 1翟明慧, 2袁殿宝, 1赵峻峰, 1张敬
1 河北北方学院附属第一医院肿瘤内科,河北 张家口 075000
2 河北北方学院附属第一医院消化内科,河北 张家口 075000
通讯: 翟明慧 Email: 1685337381@qq.com
DOI: 10.3978/j.issn.2095-6959.2019.03.002

摘要

目的:探讨PTEN基因对甲状腺髓样癌(medullary thyroid carcinoma,MTC)细胞生长、周期的影响以及作用机制。方法:使用酶切酶连的方法构建真核表达载体pcDNA3.1-PTEN,采用脂质体转染法将其转入pA10RP8细胞中。实时荧光定量PCR(real-time quantitative polymerase chain reaction, RT-qPCR)检测细胞内PTEN表达水平的变化,Western印迹法检测PTEN,Ki-67,p21,cyclin D1蛋白表达水平的变化;CCK-8法和平板克隆试验检测PTEN对pA10RP8细胞增殖的影响;细胞流式术检测pA10RP8细胞周期的变化。结果:成功构建了PTEN真核表达载体。转染pcDNA3.1-PTEN质粒的pA10RP8细胞PTEN和p21表达上调,Ki-67和cyclin D1蛋白表达下调;转染pcDNA3.1-PTEN质粒的pA10RP8细胞生长增殖受到抑制;细胞在G0/G1期比例增加,发生周期阻滞。结论:过表达PTEN能够抑制pA10RP8细胞的增殖,诱导细胞周期阻滞,其机制可能与周期蛋白p21上调和cyclin D1下调表达有关。
关键词: 甲状腺髓样癌;PTEN;细胞周期;细胞周期蛋白

Effect of over-expression of PTEN gene on growth and cell cycle of medullary thyroid carcinoma cells

Authors: 1ZHAI Minghui, 2YUAN Dianbao, 1ZHAO Junfeng, 1ZHANG Jing
1 Department of Medical Oncology, First Affiliated Hospital of Hebei North University, Zhangjiakou Hebei 075000, China
2 Department of Gastroenterology, First Affiliated Hospital of Hebei North University, Zhangjiakou Hebei 075000, China

CorrespondingAuthor: ZHAI Minghui Email: 1685337381@qq.com

DOI: 10.3978/j.issn.2095-6959.2019.03.002

Abstract

Objective: To investigate the effect of over-expression of PTEN gene on the growth and cell cycle of medullary thyroid carcinoma cells and to explore its mechanism. Methods: The eukaryotic expression vector pcDNA3.1-PTEN was constructed by restriction enzyme linked method transferred into pA10RP8 cells by liposome transfection. The mRNA expression level of PTEN in the cells were detected by RT-qPCR, the protein expression level of PTEN, Ki-67, p21 and cyclin D1 were detected by Western blot; the effect of PTEN in pA10RP8 cells on the proliferation was detected by CCK-8 and flat clones; and the changes of cell cycle were detected by flow cytometry. Results: The PTEN eukaryotic expression vector was successfully constructed; the expression of PTEN and p21 in cells with pcDNA3.1-PTEN was significantly up-regulated; the expression of Ki-67 and cyclin D1 protein in pA10RP8 cells with pcDNA3.1-PTEN was down-regulated; the proliferation of pA10RP8 cells with pcDNA3.1-PTEN was inhibited; the PTEN gene induced cell cycle arrest of pA10RP8 cells with a significant increase of the proportion of pA10RP8 cells in G0/G1 phase. Conclusion: Over-expression of PTEN could inhibit the proliferation and induce cell cycle arrest of pA10RP8 cells, whose mechanism may be related to p21 and cyclin D1 protein.
Keywords: medullary thyroid carcinoma; PTEN; cell cycle; cyclin