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Yu X.,The Affiliated Peoples Hospital of Hubei Medical College | Li Y.,The Affiliated Peoples Hospital of Hubei Medical College | Lei J.,The Affiliated Peoples Hospital of Hubei Medical College
Journal of Practical Oncology | Year: 2016

Objective: To investigate the effects of doxycycline on cell proliferation and expression of HIF-1α, HIF-2α in small cell lung cancer (SCLC) cell line H446. Methods: The inhibition of doxycycline on H446 cell proliferation was examined with MTT method. The effect of doxycycline on cell apoptotic morphological changes was observed with inverted optical microscope. Cell cycle was analyzed by flow cytometry and CellFit software. Cell apoptotic rate was detected by Annexin V-FITC fluorescence staining. Intracellular mitochondrial membrane potential was tested by Cellular Health Analysis software. The expression of the HIF-1α and the HIF-2α was analyzed by real-time PCR, and the expression of apoptosis-related proteins was examined by Western blot. Results: Doxycycline inhibited H446 cell in a time and dose dependent manner (2.5, 5, 10, 20, 40, 80 mg/L for 24 h or 48 h). Through optical microscopy, we detected that doxycycline induced the apoptosis of H446 cells in a time and dose dependent manner. Flow cytometry results showed that 10, 20, 40 mg/L doxycycline blocked H446 cell cycle to S phase after 48 h treatment, and the blockage increased as cell apoptosis rates increased, in a dose-dependent manner. Besides, in cells treated with doxycycline at 10, 20, 40 mg/L for 48 h, both the mitochondrial membrane potential and HIF expression were significantly lower (P<0.05) than those of control group. The protein levels of HIF-1α and HIF-2α were also reduced after doxycycline treatment at 10, 20, 40 mg/L for 48 h. With the increase of doxycycline concentration, the expression of the two HIF proteins further decreased (P<0.05). Conclusion: Doxycycline inhibits the proliferation of SCLC H446 cells in a time and dose dependent manner. Doxycycline blocks H446 cells to S phase, reduces mitochondrial membrane potential, down-regulates HIF-1α and HIF-2α protein expression, which are accompanied with cell apoptosis. © 2016, Editorial Board of Journal of Practical Oncology. All right reserved.


Li C.-E.,Xi'an Jiaotong University | Li C.-E.,The Affiliated Peoples Hospital of Hubei Medical College | Yao Y.-M.,Xi'an Jiaotong University | Han S.-S.,Xi'an Jiaotong University | And 2 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2012

Objective: To explore the expression and significance of transforming growth factor-β 1 (TGF-β 1) and cyclooxygenase-2 (COX-2) in benign biliary stricture. Methods: Paraffin embedded materials were analyzed for TGF-β 1 and COX-2 expression and distribution by immunohistochemical strept-avidin-biotin complex (SABC) method. Results: The positive cell percentage of TGF-β 1 and COX-2 was (71.82±13.42)% and (77.18±11.18)% in biliary stricture duct and (17.78±8.93)% and (14.69±5.69)% in normal bile duct. The positive rate of TGF-β 1 and COX-2 expression was 81.82% and 90.91% in biliary stricture duct but both 0% in normal bile duct. The immunohistochemical results showed that the expression of TGF-β 1 and COX-2 in biliary scars was significantly different from that in normal bile duct (P<0.05). TGF-β 1 and COX-2 were mainly found in fibroblasts, endothelial cells of blood vessels, and massive inflammatory cells such as macrophages. Conclusion: The formation of benign biliary stricture is associated with the increased expression of TGF-β 1 and COX-2. TGF-β 1 may promote the excretion of COX-2 and they are both involved in the formation of benign biliary stricture.

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