摘要
目的探讨上调磷酸酶及张力蛋白同源物(phosphate and tension homology deleted on chromsometen,PTEN)基因表达对三阴性乳腺癌细胞增殖及侵袭的影响及其作用机制。方法对数期人三阴性乳腺癌ZR-75-细胞株随机分为PTEN组、突变型PTEN(mutated PTEN,M-PTEN)组和对照组,PTEN组、M-PTEN组分别转染人野生型PTEN真核表达质粒(pBP-PTEN)、突变型PTEN质粒(pBP-M-PTEN),对照组未作处理。转染后24、48、72h,MTT法检测3组ZR-75-1细胞生长抑制率,转染后72h,Transwell法检测3组侵袭细胞比率,流式细胞仪检测细胞周期,Western blot法检测磷酸化局部黏着斑激酶(phosphorylated focal adhesion kinase,P-FAK)、磷酸化蛋白激酶B(phosphorylated protein kinase B,P-Akt)表达水平。结果 PTEN组PTEN蛋白相对表达量(8.49±2.92)高于M-PTEN组(0.77±0.24)(P<0.05),对照组无PTEN蛋白表达;转染后24、48、72h,PTEN组细胞生长抑制率[(27.33±21.49)%、(37.20±11.48)%、(48.11±8.14)%]均高于M-PTEN组[(5.98±1.44)%、(6.98±2.84)%、(10.83±3.11)%]和对照组[(1.09±0.33)%、(2.10±0.83)%、(4.20±2.71)%](P<0.05),M-PTEN组与对照组比较差异无统计学意义(P>0.05);转染后72h,PTEN组侵袭细胞数目[(24.29±5.82)%]低于M-PTEN组[(56.30±11.84)%]和对照组[(89.29±10.49)%](P<0.05),M-PTEN组与对照组比较差异无统计学意义(P>0.05);PTEN组G1期细胞比率[(54.89±2.67)%]低于M-PTEN组[(57.02±1.33)%]和对照组[(63.98±1.09)%],S期细胞比率[(36.56±2.10)%]高于M-PTEN组[(35.98±2.67)%]和对照组[(30.78±0.99)%](P<0.05),G2/G1期细胞比率[(1.85±0.33)%]与M-PTEN组[(1.90±1.10)%]和对照组[(1.91±1.00)%]比较差异均无统计学意义(P>0.05),M-PTEN组G1、S、G2/G1细胞比率与对照组比较差异均无统计学意义(P>0.05);PTEN组P-FAK蛋白相对表达水平(0.75±0.12)低于M-PTEN组(2.19±0.82)与对照组(3.23±0.62)(P<0.05),P-Akt蛋白相对表达(2.11±0.42)与M-PTEN组(1.83±0.74)和对照组(1.94±0.53)比较差异均无统计学意义(P>0.05);M-PTEN组P-FAK�
Objective To investigate the influence of upregulating phosphate and tension homology deleted on chromsometen （PTEN） on the proliferation and invasion of triple negative breast cancer cells and its mechanism. Methods The triple negative breast cancer ZR-75-1 cells in logarithmic phase were randomly divided into PTEN group, mutated PTEN （M-PTEN） group and control group. PTEN group and M-PTEN group were transfected by human wild type PTEN （pBP-PTEN） and mutated PTEN plasmid （pBP-M-PTEN）, while control group received no treatment. The inhibition rate was detected by MTT in 24, 48 and 72 h after transfection. The cell invasion proportion was detected by Transwell assay, the cell cycle was detected by flow cytometry, and the levels of phosphorylated focal adhesion kinase （P-FAK） and phosphorylated protein kinase B （P-Akt） were detected by Western blot in 72 h after transfection. Results The relative expression of PTEN protein was significantly higher in PTEN group （8. 49±2. 92） than that in M-PTEN group （0.77±0. 24） （P〈0.05）, and no PTEN protein was expressed in control group. In 24, 48 and 72 h after transfection, the cell growth inhibition rates were significantly higher in PTEN group （（27. 33±21. 49）%, （37. 20±11.48）%, （48.11±8. 14）%than those in M-PTEN group （（5.98±1.44）%, （6.98±2.84）%, （10.83±3. 11）% and control group （（1.09±0.33）%, （2. 10±0.83）%, （4.20±2. 71）% （P〈0.05）, and there were no significant differences between M-PTEN group and control group （P〉0.05）. In 72 h after transfection, the count of invasive PTEN cells was significantly lower in PTEN group （ （24.29 ± 5.82）%） than that in M-PTEN group （ （56.30±11.84） % ） and control group （（89. 29 ±10. 49）%） （P〈0.05）, and there was no significant difference between M-PTEN group and control group （P〉0.05）. The PTEN cell proportion in G1 phase was significantly lower in PTEN group （（54. 89 4± 2.67） %） than that in M-
作者
孙志微
戴经纬
刘丽娜
赵毅
SUN Zhi-wei;DAI Jing-wei;LIU Li-na;ZHAO Yi(Department of Pancreatic,Thyroid and Breast Surgery,Shengjing Hospital of China Medical University,Shenyang 110003,China)
出处
《中华实用诊断与治疗杂志》
2018年第8期736-739,共4页
Journal of Chinese Practical Diagnosis and Therapy
基金
沈阳市科技计划项目(F15-199-1-46)
关键词
三阴性乳腺癌
磷酸酶及张力蛋白同源物基因
局部黏着斑激酶
增殖
侵袭
Triple negative breast cancer
phosphate and tension homology deleted on chromsometen
focal adhesionkinase
proliferation
invasion
