ABSTRACT
OBJECTIVE: To study the mechanism of effects of AZD1480 on the SKOV3 ovarian cancer cell line. METHODS: The MTT method was used to assess cellular proliferation, flow cytometry for cellular apoptosis, the scratch test to determine migration, transwell chamber assays to detect cellular invasion, plate clone experiments to detect the clone forming ability and Western blotting to determine p-STAT3 protein levels. RESULTS: The proliferation rate, migration ability, invasiveness and the clone forming ability of SKOV3 cells were reduced after treatment with AZD1480, while apoptosis rate and chemotherapeutic susceptibility were increased. After treatment with AZD1480 plus cisplatin, the apoptosis rate increased significantly while the expression level of p-STAT3 protein was decreased. CONCLUSION: AZD1480 can inhibit the proliferation, invasion, metastasis and clone formation of SKOV3 cells, induce cellulsar apoptosis, increase the chemotherapeutic sensitivity and reduce the expression level of p-STAT3 protein.
Subject(s)
Apoptosis/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/pathology , Pyrazoles/pharmacology , Pyrimidines/pharmacology , Antineoplastic Agents/pharmacology , Antineoplastic Combined Chemotherapy Protocols , Blotting, Western , Cisplatin/pharmacology , Female , Flow Cytometry , Humans , In Vitro Techniques , Janus Kinase 2/antagonists & inhibitors , Tumor Cells, CulturedABSTRACT
OBJECTIVE: To investigate the effects of di-(2-ethylhexyl) phthalate (DEHP) on the expressions of Caspase-3 and Caspase-9 genes in rat Leydig cells and the apoptosis of the cells in vitro. METHODS: Leydig cells were isolated from male SD rats, primarily cultured and treated with DEHP at a low (10 nmol/L), a medium (50 nmol/L) and a high dose (100 nmol/L) for 24 hours. Then the mRNA expressions of Caspase-3 and Caspase-9 genes in the Leydig cells were detected by real time PCR, their protein expressions determined by Western blot, and the apoptosis of the Leydig cells measured by flow cytometry. RESULTS: Compared with the DMSO control group, the low-, medium- and high-dose DEHP groups showed significantly upregulated expressions of Caspase-3 mRNA (1.69 +/- 0.38 vs 3.82 +/- 0.39, 6.91 +/- 0.40 and 15.47 +/- 0.40, P < 0.05), Caspase-3 protein (0.18 +/- 0.0.09 vs 0.32 +/- 0.10, 0.61 +/- 0.08 and 0.89 +/- 0.09, P < 0.05), Caspase-9 mRNA (2.24 +/- 0.41 vs 5.16 +/- 0.43, 9.61 +/- 0.45 and 19.22 +/- 0.43, P < 0.05) and Caspase-9 protein (0.26 +/- 0.07 vs 0.40 +/- 0.08, 0.68 +/- 0.09 and 0.96 +/- 0.08, P < 0.05), as well as increased apoptosis rate of Leydig cells (4.36 +/- 1.11 vs 7.52 +/- 1.09, 12.72 +/- 1.10 and 24.59 +/- 1.11, P < 0.05), all in a dose-dependent manner. CONCLUSION: DEHP can induce the apoptosis of rat Leydig cells by activating the apoptosis Caspase pathway, and consequently affect the function of Leydig cells.
Subject(s)
Apoptosis/drug effects , Caspase 3/metabolism , Caspase 9/metabolism , Diethylhexyl Phthalate/toxicity , Leydig Cells/metabolism , Animals , Leydig Cells/drug effects , Male , Rats , Rats, Sprague-DawleyABSTRACT
AIM: To investigate the effects of eukaryotic vector expressing short hairpin RNA (shRNA) of signal transducers and activators of transcription 3 (STAT3) on the proliferation and apoptosis of SiHa cells. METHODS: shRNA templates were designed based on STAT3 gene sequence and cloned into pSilencer2.1-U6-neo vector. The resultant plasmid was transfected into SiHa cells with Lipofectamine 2000. The STAT3 protein and mRNA were detected by Western blot and RT-PCR, respectively. The cellular growth activity was assayed by MTT and the apoptosis was tested by flow cytometry. RESULTS: The plasmid pSilencer2.1-U6-neo-STAT3 was successfully constructed and transfected into SiHa cells. The expression of STAT3 in SiHa cells decreased, the cellular growth activity decreased, and the cell apoptosis increased. CONCLUSION: The siRNA expressing plasmid pSilencer2.1-U6-neo-STAT3 can inhibit cellular proliferation and induce the apoptosis of cervical cancer cells by suppressing the expression of STAT3.
Subject(s)
Apoptosis/physiology , Cell Proliferation , Genetic Vectors/physiology , RNA, Small Interfering/genetics , RNA/physiology , STAT3 Transcription Factor/metabolism , Apoptosis/genetics , Blotting, Western , Cell Line, Tumor , Female , Flow Cytometry , Genetic Vectors/genetics , Humans , RNA/genetics , Reverse Transcriptase Polymerase Chain Reaction , STAT3 Transcription Factor/genetics , Uterine Cervical Neoplasms/metabolism , Uterine Cervical Neoplasms/therapyABSTRACT
AIM: To explore the differential expression of hypoxia-inducible factor-1alpha (HIF-1alpha) in placenta tissues from pregnancy induced hypertension and normal pregnancy. METHODS: The expression of HIF-1alpha protein and mRNA was detected by Western blot and real-time PCR, respectively. RESULTS: As compared with the expression levels of HIF-1alpha protein and mRNA in placenta tissues from normal pregnancy, those from pregnancy induced hypertension increased notably (P<0.05). CONCLUSION: The high expression of HIF-1alpha in the placenta tissues pregnancy induced may relate with pathogenesis and pathophysiological process of hypertension.
