| Background and AimsCarcinoma of bladder is the frequent malignant tumor in urinary system in human body,which is the fourth malignant tumor in the world.Carcinoma of bladder threatens human health seriously.At present,great improvement about nosogenesis and treatment of carcinoma of bladder has been gained,but many questions about disease cause,diagnosis and treatment of carcinoma of bladder have not been answered.The search of molecular biology tumor markers of higher sensibility,better specificity,simple handling and less side effect is the important topic of urinary surgery.In the modern clinical medicine,there are many treatment methods of carcinoma of bladder,such as operation,radiotherapy, chemotherapy and intracavitary chemotherapeutics treatment,but the recurrence rate of carcinoma of bladder remains high.So the search of an ideal drug to prevent and treat carcinoma of bladder is important too.Survivin is a vigorous inhibitory factor against cell apoptosis.Compared with the other factors in the IAP(Inhibitor of apoptosis family of proteins,IAPs),Survivin has distinct structure and expression.And it has great value in the diagnosis,treatment and drug fast mechanism study in prevention and treatment of cancer.Survivin inhibit cell apoptosis via prevention against the activation of caspase-3 and caspase-7.The abnormality of signal transduction is the important bionomics of cell tumour.PI3K/AKT signal transduction pathway plays an important role in the maintenance of tumour cell bionomics.Tumour cell were affected by a series of of factors.PI3K/AKT signal transduction pathway is activated, the proliferation and differentiation are induced,and cell apoptosis are prevented.AKT is the key molecule in the P13K/AKT pathway,whose persistent activation has close relationship with the development of tumor.The relationship of P13K/AKT pathway and survivin deserves more investigation. The aim of this study was to observe the expression of survivin in the BTCC of human and the influence of matrine on the T24 cell,to detect the expression changes of survivin, AKT and caspase-3 of T24.And investigate the inhibition mechanism of matrine on the signal transduction pathway of survivin of T24.MethodsThis study was divided into 2 parts.In the 1st part,the specimens of 62 cases of BTCC were included into the experiment,the expression of survivin in BTCC was detected by immunohistochemistry staining,and the relationship of survivin expression with clinical pathogical characteristic was studied.In the 2nd part,cell culture technology was used. Matrine of different dosage were added into the culture fluid of T24 cell.The cell survival rate and cell apoptosis were detected.The effect of matrine on T24 cell was analysized. Immunohistochemistry staining and WB were used to detect the expression changes of survivin,AKT and caspase-3.The role of survivin on the nosogenesis and pathogenesy of carcinoma of bladder were studied.The main results and conclusions were summarized as follows:1.In 62 cases of BTCC,Survivin expression had no relationship with the gender and age of the patients.2.Survivin expressions had no relationship with the grade and infiltration of the cancer.3.Survivin expressions had no relationship with recurrence of the tumor.But Survivin expression had relationship with lymph cancerometastasis.4.In 62 cases of BTCC,Survivin expression was positive in 43 cases by immunohistochemistry staining.Survivin expression was negative in 19 cases.The positive rate was 69.4%.Survivin expression in well-differentiated grade group was positive in 8 cases.Survivin expression in moderately differentiated group was positive in 19 cases. Survivin expression in poorly differentiated group was positive in 16 cases.5.In 62 cases of BTCC,FLK1 expression was positive in 40 cases by immunohistochemistry staining.FLK1 expression was negative in 22 cases.The positive rate was 64.5%.FLK1 expression in well-differentiated grade group classification was positive in 7 cases.FLK1 expression in moderately differentiated group was positive in 17 cases.FLK1 expression in poorly differentiated group was positive in 16 cases. 6.In 62 cases of BTCC,FLT1 expression was positive in 42 cases by immunohistochemistry staining.FLT1 expression was negative in 20 cases.The positive rate was 67.7%.FLT1 expression in well-differentiated grade group classification was positive in 8 cases.FLT1 expression in moderately differentiated group was positive in 18 cases.FLT1 expression in poorly differentiated group was positive in 16 cases.7.In 62 cases of BTCC,FLK1 expression was positive in 32 cases among the survivin positive group by immunohistochemistry staining.FLK1 expression was negative in 11 cases.The expression of FLK1 and survivin had significant relationship statisticsly. FLT1 expression was positive in 33 cases among the survivin positive group by immunohistochemistry staining.FLK1 expression was negative in 10 cases.The expression of FLK1 and survivin had significant relationship statisticsly.All these results show that Survivin is an important role in the development of BTCC.Survivin,FLT-1 and FLK-1 participate in the proliferation of BTCC and angiopoiesis.8.Different dosage of matrine was used into the culture fluid of T24.In the normal group,the T24 cells were orbicular-ovate shaped or polygon shaped.Refraction was high. In matrine group,T24 cells shrinkaged,floated or even broke into pieces,which was concentration dependent and time dependent.The end of cell survival rate detection showed that matrine group at 12h was significant different from the normal group,which decreased concentration-dependently and time-dependently.In the apoptosis detection,the results had the reverse tendency,the apoptosis cell number of T24 increased concentrationdependently and time-dependently too.All these results show that matrine can inhibit T24 cells significantly.Apoptosis is one of the death ways of T24 cells.Matrine has anti-tumor effect.9.Immunohistochemistry staining and western blot detection showed the changes of survivin,AKT and caspase-3 of T24 cells.The survivin was expressed in T24 cells.In the normal group,the optical density value of survivin at 12h,24h and 48h had not any difference.In the matrine group,the optical density value of survivin decreased concentration-dependently and time-dependently.Western Blot detection showed the same results,meaning that matrine can inhibit survivin expression significantly.Western Blot detection also showed that matrine can inhibit akt expression significantly and promote the expression of caspase-3. All these results show that matrine can inhibit the survivin expression of T24 cells significantly and inhibit the activity of survivin.Matrine can inhibit the activity of akt. Matrine promote the apoptosis through the acitivation of caspase-3.Survivin plays an important role in the survival and proliferation in the T24 cells.
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