ABSTRACT
OBJECTIVE: To explore the effect of dominant negative HIF-1alpha (dn HIF-1alpha) on biological characteristics of uterine cervix cancer cell SiHa and elucidate the related mechanism. METHODS: pcDNA3. 1-dn HIF-1alpha was transfected into SiHa cells. The expression of HIF-1alpha and VEGF protein were detected by immunocytochemical method and Western Blotting. The growth proliferation of cells was surveyed by the MTT assay and cell apoptosis was detected through TUNEL after treated with CoCl2, meanwhile the results were compared with the group transfected with mock plasmid and untransfected group. RESULTS: After successfully transfected with relevant plasmid, there's no obvious difference of expression of HIF-1alpha among dn HIF-1alpha group, pcDNA3. 1 group, and untransfected group, however the expression of VEGF of dn HIF-1alpha group was significantly lower than that of the others (P < 0. 05). The proliferation ability of dn HIF-1alpha group was obviously lower than that of the other two (P < 0.05), whether it was under normoxia or chemical hypoxia induced by CoCl2. The characteristic apoptotic morphology was most significantly apparent in dn HIF-1alpha group among these three (P < 0.05). CONCLUSION: Domain negative HIF-1alpha can inhibit the proliferation of uterine cervix cancer cell and accelerate its apoptosis under hypoxia induced by CoCl2, as well as decrease the expression of VEGF protein. The implications of all this were that the domain negative HIF-1alpha may play an important role in the therapy of uterine cervix cancer.
Subject(s)
Apoptosis/physiology , Cell Proliferation , Hypoxia-Inducible Factor 1, alpha Subunit/physiology , Apoptosis/genetics , Blotting, Western , Cell Hypoxia , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/genetics , Cell Survival/physiology , Cobalt/pharmacology , Dose-Response Relationship, Drug , Female , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Immunohistochemistry , In Situ Nick-End Labeling , Plasmids/genetics , Transfection , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/metabolism , Uterine Cervical Neoplasms/pathology , Vascular Endothelial Growth Factor A/metabolismABSTRACT
OBJECTIVE: To examine the effect of Aurora A on the biological phenotypes of prostate cancer cells and the role of Aurora A in the carcinogenesis of prostate cancer. METHODS: Full length Aurora A gene was cloned and transfected into the prostate cancer cell line LNCaP to construct a positive cell clone with high expression of Aurora A. The growth and proliferation of the cells was detected by MTT. The mobility of the cells was examined by transwell cell layer infiltration assay. The growth and motility of LNCaP-Aurora A cells were compared with those of the LNCaP-pcDNA3. 1/LNCaP cells. RESULTS: Compared to the LNCaP-pcDNA3. 1/ LNCaP cells, the cells with high expression of Aurora A had faster proliferation and stronger invasive ability. Three days after cultivation, more LNCaP-Aurora A cells had grown than the LNCaP-pcDNA3. 1/LNCaP cells (P < 0.05). CONCLUSION: The increased expression of Aurora A is closely associated with the malignant phenotypes of prostate cancer. Aurora A may play a very important role in the carcinogenesis of prostate cancer.
