Wu Wanwen, Chen Ying, Huang Lan, Li Wenjian, Tao Changli, Shen Han
Guangdong Province Key Laboratory for Biotechnology Drug Candidates, School of Life Sciences and Biopharmaceutics, Guangdong Pharmaceutical University Guangzhou 510006, China.
Int J Clin Exp Pathol. 2020 Mar 1;13(3):332-346. eCollection 2020.
To investigate the effect of the AKT1 gene mutation hotspot E17K on the growth, proliferation, survival, and migration of breast cancer cells, based on the survival and prognosis of breast cancer patients with the AKT1 E17K mutation shown in TCGA database.
The survival and incidence rates of AKT1 E17K mutation hotspots in breast cancer and other cancers were extracted from the Cancer Genome Atlas (TCGA). The recombinant eukaryotic expression plasmid AKT1 E17K-pIRES2-EGFP was constructed and transfected into breast cancer MCF-7, and MDA-MB-231 cell lines. MCF-7 and MDA-MB-231 cell lines were randomly divided into blank control groups, empty plasmid groups, and recombinant plasmid groups. The growth curve was drawn using the cell counting method. The proliferation and division of breast cancer cells were detected by CFSE fluorescent dye tracking. Apoptosis was detected by Annexin V/PI double labeling and cell vitality was detected using MTT assays, and cell migratory ability was detected by cell scratch and transwell chamber tests.
In breast cancer, and other cancers, the overall survival rate of patients with an AKT E17K mutation was higher than that of patients with non-point mutation, and this mutation was the most common found in breast cancer. Compared with the wild type, the growth function of mutant MCF-7 cells was inhibited (P < 0.05), as was the proliferation of MCF-7 cells expressing the AKT1 E17K mutation gene (P < 0.001). The late apoptosis rate of mutant breast cancer cells increased (P < 0.05) and the viability was lower than that of wild-type cells (P < 0.05). Mutant MDA-MB-231 cells showed increased migration ability when compared to wild-type MDA-MB-231 cells (P < 0.05).
The expression of the AKT1 E17K mutation hotspot can inhibit the growth, proliferation, and survival ability of breast cancer cells, and promote apoptosis, while it also improves their migratory ability. The survival and prognosis of breast cancer patients with this mutation are good, which may be related to the inhibition of the PI3K/AKT/mTOR signaling pathway.
基于TCGA数据库中显示的携带AKT1 E17K突变的乳腺癌患者的生存和预后情况，研究AKT1基因突变热点E17K对乳腺癌细胞生长、增殖、存活及迁移的影响。
从癌症基因组图谱（TCGA）中提取乳腺癌及其他癌症中AKT1 E17K突变热点的生存率和发病率。构建重组真核表达质粒AKT1 E17K-pIRES2-EGFP并转染至乳腺癌MCF-7和MDA-MB-231细胞系。MCF-7和MDA-MB-231细胞系随机分为空白对照组、空质粒组和重组质粒组。采用细胞计数法绘制生长曲线。通过CFSE荧光染料追踪检测乳腺癌细胞的增殖和分裂。采用Annexin V/PI双染法检测细胞凋亡，MTT法检测细胞活力，细胞划痕和transwell小室试验检测细胞迁移能力。
在乳腺癌及其他癌症中，携带AKT E17K突变患者的总生存率高于非点突变患者，且该突变在乳腺癌中最为常见。与野生型相比，突变型MCF-7细胞的生长功能受到抑制（P < 0.05），表达AKT1 E17K突变基因的MCF-7细胞的增殖也受到抑制（P < 0.001）。突变型乳腺癌细胞的晚期凋亡率增加（P < 0.05），活力低于野生型细胞（P < 0.05）。与野生型MDA-MB-231细胞相比，突变型MDA-MB-231细胞的迁移能力增强（P < 0.05）。
AKT1 E17K突变热点的表达可抑制乳腺癌细胞的生长、增殖和存活能力，促进细胞凋亡，同时还可提高其迁移能力。携带该突变的乳腺癌患者的生存和预后良好，这可能与PI3K/AKT/mTOR信号通路的抑制有关。
