摘要目的 探讨环状RNA hsa_circ_0014130在肺腺癌细胞系中的功能和作用机制,并寻找潜在的分子抑制剂.方法 采用肺腺癌常见细胞系(PC9、H1299、A549、HCC827、BEAS-2B细胞系)进行hsa_circ_0014130的表达水平检测以及过表达和消减实验,用逆转录-即时荧光定量PCR(qPCR)验证,用细胞增殖实验和侵袭实验检测其对肺腺癌细胞的增殖和侵袭的影响,再通过数据库查询其靶向的microRNA,用免疫印迹法(Western blot)检测下游信号通路,最后使用小分子抑制剂检测对非小细胞肺癌增殖和侵袭能力的影响.结果 hsa_circ_0014130在3种细胞系中表达水平发生上调,过表达质粒和消减小干扰RNA(siRNA)均能转染到细胞中且有效.过表达hsa_circ_0014130能促进肿瘤细胞的增殖和侵袭,消减hsa_circ_0014130能够抑制肿瘤细胞的增殖和侵袭.hsa_circ_0014130能够靶向hsa-miR-566使其表达水平降低并抑制上皮间质转化.使用小分子抑制剂SB-431542并同时消减hsa_circ_0014130能够更加显著抑制肿瘤细胞的增殖和侵袭.结论 hsa_circ_0014130通过靶向hsa-miR-566进而促进TWIST1的表达来促进肺癌细胞的侵袭和增殖,消减其表达水平同时与小分子抑制剂SB-431542联用能显著抑制肺癌细胞的增殖和侵袭,这提示hsa_circ_0014130是潜在的肺癌治疗靶点.

abstractsObjective To investigate the function and mechanism of hsa_circ_0014130 in lung adenocarcinoma cell line and to find potential molecular inhibitors. Methods The hsa_circ_0014130 expression level detection and overexpression and subtraction experiments were performed using common cell lines of lung cancer (PC9, H1299, A549, HCC827, and BEAS-2B). qPCR was used to verify the proliferation and invasion of lung cancer cells by MTS and invasion assay, and then the targeted microRNA was searched through the database. Western blot was used to detect the downstream signaling pathways, and finally the effect of small molecule inhibitors was investigated on proliferation and invasion of non-small cell lung cancer. Results The expression level of hsa_circ_0014130 was up-regulated in the three cell lines, and both the overexpression plasmid and the subtractive siRNA were effectively transfected into the cells. Overexpression of hsa_circ_0014130 was able to promote the proliferation and invasion of tumor cells, and knockdown of hsa_circ_0014130 inhibited the proliferation and invasion of tumor cells. hsa_circ_0014130 was capable to target hsa-miR-566 to reduce its expression level and to inhibit epithelial-to-mesenchymal transition. The use of the small molecule inhibitor SB-431542 and simultaneous reduction of hsa_circ_ 0014130 significantly inhibited the proliferation and invasion of tumor cells. Conclusions The hsa_circ_0014130 promotes the invasion and proliferation of lung cancer cells by targeting hsa-miR-566 to enhance the expression of TWIST1, and its expression level can be significantly inhibited by the small molecule inhibitor SB-431542, which significantly inhibits the proliferation and invasion of lung cancer cells. Therefore,hsa_circ_0014130 is a potential lung cancer treatment target.