ABSTRACT
OBJECTIVE：To study t he effects of bro mophenylcurcumin（GL63）on the apoptosis ，migration and invasion of human cholangiocarcinoma RBE cells ，and to investigate its mechanism based on JAK/STAT signaling pathway. METHODS ：MTT assay was used to detect the effects of different concentrations of GL 63 [0（blank control ，similarly hereinafter ），1.25，2.5，5，10， 20，40 μ mol/L] on the proliferation of RBE cells after 48 h treatment ；the IC 50 was calculated. The effects of different concentrations of GL 63（0，5，10，20 μmol/L）on colony formation were detected by crystal violet staining after 48 h treatment. Flow cytometry ，Hoechst 33342 staining，cell scratch test and Transwell chamber invasion test were used to detect the effects of different concentrations of GL 63（0，5，10，20 μmol/L）on cell cycle distribution ，apoptosis，migration and invasion ability after 24 h treatment. Western blotting assay was adopted to detect the effects of different concentration of GL 63（0，5，10，20 μmol/L） on the expression of JAK 2/STAT3 signal pathway associated proteins. RESULTS ：The proliferation inhibition rates of RBE cells in different concentrations of GL 63 groups（1.25-40 μmol/L）were significantly increase d，compared with blank control group （P＜ 0.01），and showed a dose-dependent trend ，with IC 50 of （8.46±1.30）μmol/L. Compared with blank control group， 85917439。E-mail：zhaoji-an-88@163.com inhibition rates of RBE cell colony formation were significantly decreased in different concentrations （5，10，20 μmol/L）of GL 63 groups（P＜0.01）. The percentage of RBE cells at G 0/G1 phase increased significantly ，while that at S phase decreased significantly （P＜0.01）. The apoptotic rate increased significantly（P＜0.01），and the nucleus showed dense pyknosis and apoptotic bodies. The rate of cell migration and healing was significantly decreased （P＜0.01），and the number of invasive cells through basement membrane was significantly decreased （P＜ 0.01）. The protein expression of p-JAK 2， p-STAT3， Bcl-2， MMP-2， MMP-9， Pro-caspase-9 and P ro-caspase-3 were down-regulated significantly while the expression of Bax ，Cyt-c，Cleaved-caspase-9 and Cleaved-caspase- 3 were up-regulated significantly（P＜0.01）. CONCLUSIONS ：GL63 may inhibit the proliferation ，migration and invasion of RBE cells and promote its apoptosis by inhibiting JAK 2/STAT3 signal pathway.
ABSTRACT
Objective To explore the effect of luteolin on the proliferation of osteosarcoma stem cells.Methods CD133 +osteosarcoma stem cells were separated from MG63 cells by flow cytometer.MTT was used to investigate the effects of luteolin(0,0.01,0.02,0.04 mg/mL)on the proliferation of osteosarcoma stem cells.Western blot was used to detect the levels of Ki67 protein and components of JAK2/STAT3 signal pathway in osteosarcoma stem cells induced.Results After sorting,the content of the CD133 +fraction was enriched up to(87.60 ±5.06)%.MTT assay showed that,compared with the control group,luteolin(0.01,0.02,0.04 mg/mL)inhibited proliferation of CD133 + osteosarcoma stem cells(P <0.05).Western blot also showed that luteolin significantly decreased the level of Ki67 compared with the control group(P<0.05).In addition,the luteolin inhibited the expression of p-JAK2 and p-STAT3 in JAK2/STAT3 signal pathway of CD133 + osteosarcoma stem cells compared with the control group ( P <0.05 ) . Conclusion Luteolin might be a suppressor of osteosarcoma stem cells.
ABSTRACT
Aim To investigate the inhibitory effect of Evodiamine on JAK2/STAT3 signal pathway in human colorectal cancer cell line HCT-116 . Methods Cells were cultured with 6. 0 μmol·L-1 Evodiamine for 2, 4 and 6 h, respectively. Cell nuclear morphology was detected by Hoechst staining and protein expression levels of JAK2 , p-JAK2 , STAT3 and p-STAT3 were examined by Western blot. Cells were treated with dif-ferent concentrations of AG490 for 48 h to select proper working concentration and cells treated with 6 μmol · L-1 EVO and 50 μmol · L-1 AG490 to compare the modulatory effect of EVO with AG490 on JAK2/STAT3 signal pathway. Results Hoechst staining revealed that Evodiamine could induce cells apoptosis, chroma-tin condensation gathered and typical apoptotic mor-phological changes in a time-dependent manner;West-ern Blot suggested that EVO could inhibit p-STAT3 significantly. After treatment with AG490, JAK2/STAT3 signal pathway was inactivated, the inhibitory effect of EVO on p-STAT3 was stronger than that of AG490 , while EVO combined with AG490 could fur-ther inhibit the expression of p-STAT3 significantly. Conclusions The anticancer effect of Evodiamine is mainly mediated by the modulation of JAK2/STAT3 signal pathway in HCT-116 cells.
ABSTRACT
AIM: To investigate the correlation of hepatitis B virus X protein (HBx) with renal tubular epithelialcell apoptosis in hepatitis B virus-associated glomerulonephritis (HBVGN) and the possible signaling mechanism. METHODS: The activation of JAK2/STAT3 signal pathway and the expression of apoptosis -related proteins in humankindey proximal tubular epithelial cells (HK-2 cells) were determined by Western blotting after transfection with HBx eukaryoticexpression vector.The cell proliferation was observed by CCK-8 assay.The cell apoptosis was analyzed by the imagingof HO33342 staining, transmission electron microscopy and flow cytometry with Annexin V /PI double staining.RESULTS:After transfection of the target gene HBx, the expression levels of both p-JAK2 and p-STAT3 were significantly increased.At the same time, the cell proliferation was obviously inhibited, and the apoptotic rate was increased.After incubationwith AG490, the JAK2/STAT3 signal pathway was partially blocked, and the cell apoptosis induced by HBx was reduced. CONCLUSION: HBx up-regulates the activation of JAK2/STAT3 signal pathway to induce renal tubular epithelialcell apoptosis, which is possibly involved in the pathogenic mechanism that HBV directly damages nephridial tissue .
ABSTRACT
Objective: To study the effects of serum containing Dahuang Fuzi (Rhubarb and Aconite) Decoction (DFD) on JAK2/STAT3 signal pathway in mice with severe acute pancreatitis (SAP). Methods: SAP model in mice was constructed, and then the peritoneal macrophages were vaccinated into the culture plate to set model group, serum containing DFD groups (2.5%, 5%, and 10%), AG490 (10 μmol/L ) positive group, and peritoneal macrophages of normal mice acted as normal control group. After an incubation of 2 h, cells were added with serum containing DFD at different concentration or AG490 0.5 mL, 24 h later, the concentration of TNF-α and IL-6 in supernatant were determined by quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kits, mRNA and protein expression levels of JAK2 and STAT3 in cells were evaluated by Western blotting. Results: DFD could significantly decrease the levels of cytokines TNF-α and IL-6 in the supernatants, inhibit the mRNA and protein expression levels of JAK2 and STAT3 in peritoneal macrophages of SAP mice. Conclusion: DFD could inhibit the JAK2/STAT3 signal pathway and inflammatory responses in peritoneal macrophages of SAP mice.