Xiong Guosheng, Chen Xiaobo, Zhang Qi, Fang Yu, Chen Wanling, Li Caixia, Zhang Jing
Department of Thoracic Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan, China.
Department of Pharmacy, The First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan, China.
Biochem Biophys Res Commun. 2017 Apr 8;485(3):627-634. doi: 10.1016/j.bbrc.2017.02.013. Epub 2017 Feb 9.
The objective of this study was to construct a recombinant vector expressing siRNA targetedly inhibiting aquaporin 3 (AQP3), and to evaluate the effects of AQP3 inhibition on the proliferation and invasion of XWLC-05 human lung cancer cells.
We obtained human AQP3 sequence from the Genbank and established the recombinant vector expressing siRNA targeting AQP3. After the transfection of the recombinant vectors, the expression of AQP3 was determined by RT-PCR and western blot. The MTS assay, flow cytometry and Transwell assay were conducted to detect the proliferation, cell cycle process, apoptosis and invasion of XWLC-05 cells. Then the activity of metal matrix proteinase (MMP) 2 was determined by gelatin zymography. Tumor formation in vivo experiments were also conducted in nude mice.
RNA interference (RNAi) of AQP3 substantially suppressed the XWLC-05 cell proliferation and invasion, blocked the cell cycle progressing and promoted cell apoptosis. In addition, the activity of MMP2 was remarkably attenuated in RNAi group. AQP3 RNAi did not affect the tumor formation rate in nude mice but reduced the tumor growth.
The inhibition of AQP3 retarded the growth and invasiveness of XWLC-05 lung cancer cells and decreased the activity of MMP2.
本研究旨在构建靶向抑制水通道蛋白3（AQP3）的小干扰RNA（siRNA）表达重组载体，并评估抑制AQP3对XWLC-05人肺癌细胞增殖和侵袭的影响。
从基因库获取人AQP3序列，构建靶向AQP3的siRNA表达重组载体。转染重组载体后，采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测AQP3的表达。通过噻唑蓝比色法（MTS法）、流式细胞术和Transwell实验检测XWLC-05细胞的增殖、细胞周期进程、凋亡及侵袭情况。然后用明胶酶谱法检测基质金属蛋白酶（MMP）2的活性。还在裸鼠体内进行了肿瘤形成实验。
AQP3的RNA干扰（RNAi）显著抑制XWLC-05细胞的增殖和侵袭，阻断细胞周期进程并促进细胞凋亡。此外，RNAi组中MMP2的活性明显减弱。AQP3 RNAi不影响裸鼠的肿瘤形成率，但可降低肿瘤生长速度。
抑制AQP3可延缓XWLC-05肺癌细胞的生长和侵袭，并降低MMP2的活性。
