ABSTRACT
We investigated the inhibitory effect and mechanism of action of bruceantin (BCT) on the proliferation, invasion and migration of non-small cell lung cancer (NSCLC) cells. The cytotoxic activity of BCT was measured by MTT assay; a colony forming assay, wound healing assay, and a Transwell assay were used to investigate the anti-proliferative, anti-migration, and anti-invasion effects, respectively; immunoblotting and RT-qPCR were used to detect the expression of related proteins, miRNA, and mRNA, respectively, that were involved in cell proliferation, migration, and invasion. Two gene prediction websites were used to predict the downstream target gene of miRNA. Our results show that BCT has a potent cytotoxic effect on NSCLC cell lines, with a half maximal inhibitory concentration (IC50) of BCT against H1299, PC-9, and A549 of 0.12 ± 0.02, 0.31 ± 0.20, and 2.07 ± 0.70 μmol·L-1, respectively. When H1299 cells were treated with 0.03, 0.15, and 0.75 μmol·L-1 BCT for 24 h, the proliferation, migration, and invasive ability were inhibited in a concentration-dependent manner. It is worth noting that the expression level of miRNAs related to cell migration and invasion, such as miR-29a-3p, miR-21-3p, miR-183-5p, and miR-34b-5p increased with the concentration of BCT, especially for miR-29a-3p. Using the two gene prediction websites, we predict that integrin β1 (ITGB1) may be the target gene of miR-29a-3p; immunoblot results further show that a variety of proteins related to cell proliferation, migration, and invasion, such as various proteins of the integrin family, β-catenin, p-Src, and vascular endothelial growth factor, all decreased in a concentration-dependent manner, among which the reduction of ITGB1 protein was the most obvious. RT-qPCR results showed that there was no change in ITGB1 mRNA expression. We speculate that BCT might inhibit the expression of ITGB1 protein by up-regulating miR-29a-3p independent of its mRNA level. The in-depth mechanism needs to be further explored. This study suggests that BCT has the potential for further development in the treatment of NSCLC.
ABSTRACT
Objective:To evaluate the regulation effect of corn silk on fatty acid compositions in adipose tissues of rats with type 2 diabetes mellitus（T2DM） based on gas chromatography-mass spectrometry （GC-MS）， and explore the mechanism of corn silk water decoction for lipid metabolism disorder in . Method:Rat model of T2DM was established by high-sugar and high-fat diet combined with intraperitoneal injection of 35 mg·kg-1 streptozocin （STZ）. They were randomly divided into model group，metformin group，and low，medium and high dose groups（5.4，10.8，21.6 g·kg-1） of corn silk decoction， with administration of 4 weeks. Serum was taken from the abdominal aorta for the determination of free fatty acids （FFAs），blood lipids ［total cholesterol（TC），triglyceride（TG），high-density lipoprotein（HDL-C），low density lipoprotein（LDL-C）］，and adipose tissue was taken for pathological section analysis. At the same time，GC-MS technology and multivariate statistical analysis methods were adopted to investigate the fatty acid compositions in adipose tissue. Result:After treatment with corn silk decoction，the contents of FFAs，TC，TG，HDLC and LDLC in T2DM rats were all reversed. As compared with the model group，the morphology of rat adipocytes，macrophage infiltration，vascular congestion and other phenomena were restored after administration. GC-MS analysis showed that the contents of palmitic acid and stearic acid in the adipose tissue of T2DM rats were significantly increased，while the contents of tetradecanoic acid，pentadecanoic acid，9-octadecenoic acid，9-hexadecenoic acid and 5，8，11，14-eicosatetraenoic acid were decreased significantly，and all the dose groups of corn silk had a reversing effect on differentially expressed fatty acids. Conclusion:Corn silk can be able to reverse the fatty acid content in adipose tissue and restore the morphology of adipose tissue，with a regulating effect on the lipid metabolism disorder in T2DM rats.