No abstract

Hepatocellular carcinoma (HCC) is one of the most common tumor malignances with poor chemotherapeutic efficiency due to chemoresistance. MicroRNAs (miRNAs) have essential roles in regulating chemoresistance. However, the mechanism underlying the involvement of miR-212-3p in paclitaxel (PTX) resistance in HCC remains unclear. PTX resistance was investigated in the present study by assessing cell viability, the half maximal inhibitory concentration of PTX, resistance-associated protein levels and apoptosis. The expression levels of miR-212-3p and zinc finger E-box binding homeobox 2 (ZEB2) were detected by reverse transcription-quantitative PCR and western blotting. The epithelial-mesenchymal transition (EMT), migration and invasion were evaluated by western blotting and transwell assay. The association between miR-212-3p and ZEB2 was investigating by the luciferase activity. The results showed that treatment of HCC cells with PTX inhibited cell viability and miR-212-3p level. Moreover, miR-212-3p was reduced and its overexpression resulted in decreased cell viability, half maximal inhibitory concentration (IC 50) of PTX and levels of P-glycoprotein and glutathione S-transferase π, but increased cell apoptosis, in Huh7/PTX cells. However, miR-212-3p knockdown induced opposite effects in Huh7 cells. Furthermore, EMT, migration and invasion were induced in Huh7/PTX cells and the addition of miR-212-3p inhibited EMT, migration and invasion. Meanwhile, miR-212-3p abrogation caused the opposite effects in Huh7 cells. Additionally, ZEB2 was directly targeted by miR-212-3p and its restoration or silencing abated the effect of miR-221-3p overexpression or knockdown in Huh7/PTX or Huh7 cells, respectively. The data from the present study suggest that miR-212-3p attenuates PTX resistance, by regulating EMT, migration and invasion via targeting ZEB2 in HCC cells, indicating a novel target for HCC chemotherapy.

Carthamus tinctorius, used in traditional Chinese medicine, has many pharmacological effects, such as anticoagulant effects, antioxidant effects, antiaging effects, regulation of gene expression, and antitumor effects. However, there is no report on the antiallergic effects of the components in C. tinctorius. In the present study, we investigated the antiallergic components of C. tinctorius and its mechanism of action. A rat basophilic leukemia 2H3/cell membrane chromatography coupled online with high-performance liquid chromatography and tandem mass spectrometry method was developed to screen antiallergic components from C. tinctorius. The screening results showed that Hydroxysafflor yellow A, from C. tinctorius, was the targeted component that retained on the rat basophilic leukemia 2H3/cell membrane chromatography column. We measured the amount of β-hexosaminidase and histamine released in mast cells and the key markers of degranulation. The release assays showed that Hydroxysafflor yellow A could attenuate the immunoglobulin E induced release of allergic cytokines without affecting cell viability from 1.0 to 50.0 μM. In conclusion, the established rat basophilic leukemia 2H3 cell membrane chromatography coupled with online high-performance liquid chromatography and tandem mass spectrometry method successfully screened and identified Hydroxysafflor yellow A from C. tinctorius as a potential antiallergic component. Pharmacological analysis elucidated that Hydroxysafflor yellow A is an effective natural component for inhibiting immunoglobulin E-antigen-mediated degranulation.

MaiLuoNing injection is a traditional Chinese medicine that used clinically since the 1950s in China. However, anaphylactic reactions, through the potentiation of mast cell degranulation, have been reported. In the present study, a rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization-ion trap-time of flight-mass spectrometry method was established for screening, analyzing, and identifying the potential anaphylactic components of MaiLuoNing injection. Harpagoside, a potential degranulator of rat basophilic leukemia-2H3 cells, was retained in rat basophilic leukemia-2H3 cell membrane chromatography. We aimed to evaluate the retained components to determine which of those were capable of inducing degranulation of basophilic leukemia cells. A β-hexosaminidase assay revealed that harpagoside can induce rat basophilic leukemia-2H3 cell degranulation in a dose-dependent manner. BLBA/c mice also exhibit passive cutaneous anaphylaxis in response to harpagoside. These results indicate that rat basophilic leukemia-2H3 cell membrane chromatography coupled with high-performance liquid chromatography and electrospray ionization ion trap time-of-flight mass spectrometry is effective in screening for the anaphylactic components of MaiLuoNing injection.