Abstract：
This study investigated whether microbubbles activated by low-frequency ultrasound enhanced the anti-tumor effects of curcumin in glioma cells. CCK8 proliferation assay, scratch migration assay, and transwell invasion assay were performed to estimate the proliferation, migration, and invasion rates of the glioma cells in blank control and different treatment groups, respectively. Quantitative RT-PCR (qRT-PCR) analysis was performed to determine the relative expression levels of VEGF and NCAM mRNAs in the various experimental groups. Western blotting was performed to determine the activity status of the TGF-β1/Smad signaling pathway in various groups of glioma cells by estimating the expression levels of p-SMAD2/3, VEGF, and NCAM proteins. Combined treatment (Cur-Us-MBs) with microbubbles activated by low-frequency ultrasound and curcumin significantly reduced the in vitro proliferation, migration, and invasiveness of glioma cells compared to the control and other treatment groups. Furthermore, Cur-Us-MBs significantly reduced the expression levels of VEGF and NCAM mRNAs and proteins and p-Smad2/3 proteins , including those cells stimulated with rhTGF-β. These suggested that microbubbles activated by low-frequency ultrasound enhanced the inhibition of TGF-β1/Smad/VEGF/NCAM signaling pathway by curcumin，and enhanced the antitumor effects of curcumin by significantly reducing in vitro proliferation, migration, and invasiveness of glioma cells through this pathway.
摘要：
这项研究调查了低频超声激活的微泡是否增强了姜黄素在神经胶质瘤细胞中的抗肿瘤作用。CCK8增殖试验,划痕迁移试验,和transwell侵袭试验进行评估增殖，迁移,空白对照组和不同治疗组胶质瘤细胞的侵袭率,分别。进行定量RT-PCR(qRT-PCR)分析以确定各种实验组中VEGF和NCAMmRNA的相对表达水平。Westernblotting通过估计p-SMAD2/3、VEGF、和NCAM蛋白。通过低频超声和姜黄素激活的微泡联合治疗（Cur-Us-MBs）显着降低了体外增殖，迁移,与对照组和其他治疗组相比，胶质瘤细胞的侵袭性。此外，Cur-Us-MB显著降低VEGF和NCAMmRNA及蛋白和p-Smad2/3蛋白的表达水平,包括用rhTGF-β刺激的细胞。提示低频超声激活的微泡增强了姜黄素对TGF-β1/Smad/VEGF/NCAM信号通路的抑制作用，并通过显著降低体外增殖来增强姜黄素的抗肿瘤作用。迁移,和通过这个途径侵袭神经胶质瘤细胞。
