Inhibitory Effect and Mechanism of Jingulian Capsule on Human Breast Cancer MDA-MB-231 Cells / 中国实验方剂学杂志

Objective:To observe effect of Jingulian capsule on the proliferation of human breast cancer MDA-MB-231 cells and investigate its action mechanism against triple negative breast cancer （TNBC）. Method:The ingredients of Jingulian capsule were identified by ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. The inhibitory effect of Jingulian capsule at different doses （0.125，0.25，0.5，1，and 2 g·L^-1） against the proliferation of MDA-MB-231 cells were detected by methyl thiazolyl tetrazolium （MTT） assay. After treatment for 24 h， the morphological changes in nuclear apoptosis of MDA-MB-231 cells were detected by Hoechst 33258 staining. The effect of different concentrations of Jingulian capsule on the apoptosis and cycle of MDA-MB-231 cells after different treatment time were determined by flow cytometry. The protein expression levels of Poly-ADP-ribose polymeras （PARP）， proto-oncogene c-Myc， cyclin B₁， and phosphorylated extracellular signal-regulated kinase （p-ERK） in each group were assayed by Western blot. Result:A total of 113 compounds in Jingulian capsule were identified by UPLC-MS/MS. As revealed by MTT assay，compared with blank group，Jingulian capsule （0.125，0.25，0.5，1，2 g·L^-1） significantly inhibited viability of MDA-MB-231 cells （<italic>P</italic><0.01）， with the half maximal inhibitory concentration （ IC₅₀） of（0.13±0.02）g·L^-1. According to flow cytometry，compared with the blank group，Jingulian capsule at 1 g·L^-1 significantly induced the apoptosis of MDA-MB-231 cells （<italic>P</italic><0.05）and Jingulian capsule at 0.5， 1 g·L^-1 obviously increased the number of MDA-MB-231 cells in S phase （<italic>P</italic><0.05，<italic>P</italic><0.01）. The results of Western blotting demonstrated that the protein expression levels of PARP，c-Myc，and cyclin B₁ in 0.5， 1 g·L^-1Jingulian capsule groups were remarkably down-regulated as compared with those in the blank group（<italic>P</italic><0.01）， and the protein expression level of p-ERK in 1 g·L^-1Jingulian capsule group was also down-regulated （<italic>P</italic><0.01）. Conclusion:Jingulian capsule is able to inhibit the proliferation of MDA-MB-231 cells，induce S phase cell cycle arrest， and promote their apoptosis， which may be related to the inactivation of the MAPK signaling pathway.

Effects and mechanism of bakuchiol-induced S-phase cell cycle arrest in human breast cancer MCF-7 cells / 中草药

Objective: To investigate the cell growth inhibitory effect and molecular mechanism of bakuchiol against human breast cancer MCF-7 cells. Methods: The growth inhibitory effect of bakuchiol on MCF-7 cells was tested by MTT assay. Flow cytometry was used to investigate the distribution of cell cycle and ROS generation. Fluorescence microscope was used to observe the change of cell nucleus. Western blotting was used to detect the expression of the protein related to cell cycle and MAPK family. The ROS scavenger and inhibitors of MAPK family were introduced to investigate the effect on the growth inhibitory rate and the levels of cell cycle related protein by bakuchiol. Results: Bakuchiol inhibited the cell growth on the MCF-7 cells in dose- and time-dependent manner, which showed stronger effect than that of 5-fluorouracil. Furthermore, bakuchiol induced S-phase arrest in MCF-7 cells via ROS generation. The production of ROS up-regulated p-p53 and p21 expression, and then decreased CDK2 and CyclinA2. The changes of bakuchiol on these proteins could be reversed by the ROS scavenger Trion, indicating that ROS was associated with bakuchiol-induced S-phase arrest. In addition, pretreatment with p38MAPK inhibitor SB203580 decreased bakuchiol-caused ROS generation, suggesting that the production of ROS was dependent on p38MAPK pathway. Conclusion: The proliferation inhibitory effect of bakuchiol on MCF-7 cells is related with S-phase cell cycle arrest, and ROS plays a role in the bakuchiol-induced S-phase arrest.

Investigation into perturbed nucleoside metabolism and cell cycle for elucidating the cytotoxicity effect of resveratrol on human lung adenocarcinoma epithelial cells / 中国天然药物

In an effort to understand the molecular events contributing to the cytotoxicity activity of resveratrol (RSV), we investigated its effects on human lung adenocarcinoma epithelial cell line A549 at different concentrations. Cellular nucleoside metabolic profiling was determined by an established liquid chromatography-mass spectrometry method in A549 cells. RSV resulted in significant decreases and imbalances of deoxyribonucleoside triphosphates (dNTPs) pools suppressing subsequent DNA synthesis. Meanwhile, RSV at high concentration caused significant cell cycle arrest at S phase, in which cells required the highest dNTPs supply than other phases for DNA replication. The inhibition of DNA synthesis thus blocked subsequent progression through S phase in A549 cells, which may partly contribute to the cytotoxicity effect of RSV. However, hydroxyurea (HU), an inhibitor of RNR activity, caused similar dNTPs perturbation but no S phase arrest, finally no cytotoxicity effect. Therefore, we believed that the dual effect of high concentration RSV, including S phase arrest and DNA synthesis inhibition, was required for its cytotoxicity effect on A549 cells. In summary, our results provided important clues to the molecular basis for the anticancer effect of RSV on epithelial cells.

Effect of sodium arsenite on cell cycle of SH-SY5Y cells and intervention of MPST over-expression / 中国药理学通报

Aim To investigate the effects of sodium arsenite (NaAsO2) on the cell cycle and growth, and the intervention of MPST over-expression in the neural cells. Methods NaAsO2 was used to treat SH-SY5Y neuroblastoma cells for 48 h from the blank control (BC), empty vector control ( transfected with empty vector, NC ) and over-expression group ( lentiviral transfection with MPST,OP). The methods of CCK-8, crystal violet staining,flow cytometry and Western blot were used to examine cell viability,adherent rate,cell cycle and protein expression of p53,CDC25A,CyclinA and CDK2. Results The cell viability and adherent rate significantly decreased after treatment with NaAsO2 for 48 h,which was reversed in OP group (P <0. 01). Meanwhile, NaAsO2 also significantly increased the proportion of S phase cells and p53 protein expression, and down-regulated the protein levels of CDC25A,Cyc-lin A and CDK2 in BC and NC groups ( P < 0. 01), whereas the above changes of protein levels were significantly antagonized in OP group compared with NC group (P < 0. 05, P < 0. 01). Conclusions NaAsO2 inhibits the cell growth by inducing S-phase arrest and over-expression of MPST could reverse the noxious effects caused by NaAsO2 in SH-SY5Y cells.

Experimental study on the pancreatic cancer G1/S phase arrest byβ2-adrenergic antagonist / 西安交通大学学报(医学版)

Objective To investigate the effects of β2-adrenergic antagonist ICI118 ,551 on pancreatic cancer cell G1/S phase arrest and its action mechanism .Methods The cell cycle indexes were determined by the flow cytometry assay ;the expressions of Cyclin D1 and Cyclin E were analyzed by Western blot ;the activation of NF-κB was measured by electrophoretic mobility shift assay ;the proliferation of PanCa cells was determined by BALB/c athymic nude mice subrenal capsular assay .Results β2-adrenergic antagonist ICI118 ,551 significantly induced G1/S phase arrest compared with β1-adrenergic antagonist metoprolol in MIA PaCa-2 and BxPC-3 cell lines .ICI118 ,551 inhibited the expressions of Cyclin D1 and Cyclin E and reduced the activation of NF-κB .The proliferation of PanCa cells was strongly suppressed in the renal capsule xenografts in mice after ICI 118 ,551 treatment .Conclusion The blockage ofβ2-adrenoceptor markedly induces PanCa cells to arrest at G1/S phase and inhibits the proliferation of pancreatic cancer cells .