长链非编码GATS表达干扰质粒构建抑制乳腺癌细胞增殖和侵袭的研究

Inhibitory regulation of human long non-coding RNA GATS on proliferation and invasion of breast cancer cells

摘要:

目的  探讨长链非编码GATS对乳腺癌细胞增殖和侵袭的影响。方法  构建的人LncRNA-GATS-
shRNA1～4慢病毒载体转染乳腺癌MDA-MB-231细胞后，qPCR筛选出干扰效率为50.0%和54.0%的sh1组和sh3组，MTT法和Transwell实验分别检测转染后细胞增殖和侵袭能力；流式细胞术检测转染sh1组慢病毒后细胞周期分布及凋亡变化。结果  测序证实，LncRNA-GATS的shRNA寡聚核苷酸序列已被克隆到pLV-GATS载体；转染MDA-MB-231细胞后，与阴性对照组比较，sh1组和sh3组的乳腺癌细胞增殖和侵袭能力均降低（P <0.01），以sh1组更显著；sh1组细胞周期被阻滞于S期（P <0.01）；细胞凋亡率无明显变化（P > 0.05）。结论  LncRNA-GATS可能下调乳腺癌细胞侵袭力，并通过抑制细胞周期进展降低细胞增殖能力。

Abstract:

Objective To investigate the effect of human long non-coding RNA GATS on proliferation and invasion of breast cancer MDA-MB-231 cells. Methods Four human GATS-lncRNA-specific oligonucleotide sequences were designed and synthesized previously, then were inserted into the pLV-GATS vector. After infecting the MDA-MB-231 cells, transfection efficiencies of GATS-lncRNA-shRNA1 and GATS-lncRNA-shRNA3 were analyzed and screened by qPCR, which were 50.0% and 54.0% respectively. MTT and Transwell assays were used to detect the proliferation and invasion ability of the transfected cells, respectively. Flow cytometry was used to search the changes of cell cycle distribution and apoptosis of the MDA-MB-231 cells transfected GATS-lncRNA-shRNA1 lentiviral vector. Results Compared with the cells transfected with scrambled shRNA, the proliferation and invasion ability of cells the sh1 group and sh3 group was inhibited, especially t-hose  the sh1 group. Flow cytometry showed the sh1 group cell cycle was arrested in S phase, and the cell apoptosis rate had no significant change (P > 0.05). Conclusions lncRNA GATS inhibits the invasion of breast cancer cells, and down regulates the proliferation of tumor cells through arresting cell cycle in S phase.