| The first partObjective To investigate the expression of TGF-β1 and Smad4 protein in cervical squamous cell carcinoma(CSCC) and the relationship with clinical pathology.Methods Immunohistochemical staining of TGF-β1 and Smad4 protein was performed in 89 cases of CSCC and 19 cases of normal cervical epithelium tissue(NCE) paraffin sections, and analyze the relationship between the expression and clinical pathology of CSCC, correlation between the TGF-β1 and Smad4 protein in all above tissues.Results The positive rate of the TGF-β1 protein expression in CSCC was 71.9% and higher than the rate in NCE(31.1%)(P<0.01). TGF-β1 had closely relative to pathological differentiation and infiltration(P<0.05) of CSCC, but not clinical staging, age and lymph-node metastases(P>0.05). The positive rate of the Smad4 protein expression in CSCC was 32.6% and lower than the rate in NCE(100%)(P<0.01). Smad4 had closely relative to pathological differentiation of CSCC, but not clinical staging, infiltration, age and lymph-node metastases(P>0.05); There was no significant correlation between the expression of the TGF-β1 protein and the Smad4 protein (r=0.11, P>0.05).Conclusions Both TGF-β1 and Smad4 protein had closely relative to pathological differentiation of CSCC, they are anticipated to be markers of malignant degree of CSCC. TGF-β1 protein plays an important role in growth and invasion in CSCC.The second partObjective To study the effect of ursolic acid(UA) on human cervical squamous cell SiHa and its molecular mechanisms.Methods The inhibitory effect of UA on the proliferation of SiHa cells was determined by MTT method. Wright-Giemasa dyeing, Hoechst 33258 fluorescent dyeing and electron microscope were used to observe the morphological changes. Cell cycle and apoptosis rate were analysed by flow cytometry (FCM), The expressions of TGF-β1 mRNA and Smad4 mRNA involved in the effect of UA on SiHa cells were studied by RT-PCR.Results UA strongly inhibited the growth of SiHa cells in a dose and time-dependent manner. The morphological changes of apoptosis, such as nuclear fragmentation or condensed particles were identified after incubation with UA by Wright-Giemasa dyeing and Hoechst 33258 fluorescent dyeing. Cytoplasm and nuclear condensation were observed by electron microscope in UA-treated cells. Cell cycle analysis revealed that SiHa cells treated by UA were blocked in G0/G1 phase. The apoptotic peak was found with flow cytometry. The expression of TGF-β1 mRNA decreased and Smad4 mRNA increased in a time-dependent manner in UA-treated cells.Conclusions UA had the effect of inhibition on SiHa cells proliferation and induction of cell apoptosis. The cell cycle was blocked in G0/G1 phase. The mechanisms perhaps related to the TGF-β/Smads signaling pathway, which was regained through up-regulated Smad4 mRNA by UA.
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