| Objective: This study intends to detect the expression of CCND1 and Wnt / ?-catenin signaling pathway-related molecules in clinical tissues;to detect the expression of CCND1 and Wnt / ?-catenin signaling pathwayrelated molecules in cervical cancer HeLa cells at the cellular level,and further use Scutellaria baicalensis Intervene in its cell line,observe the changes of the above-mentioned related factors,and initially clarify the role of baicalein in the cell cycle;block the Wnt / ?-catenin signaling pathway by pathway blockers,and observe the effect of baicalein.Further clarify its path of action;construct an animal model of cervical cancer,intervene with baicalein,detect the expression of CCND1 and Wnt / ?-catenin signaling pathway-related molecules,and further clarify the inhibitory effect of baicalein on cervical cancer invasion and metastasis.Methods: 50 cases of cervical cancer biopsy and 46 cases of adjacent cancer tissues were selected.All patients met the diagnostic criteria for cervical cancer and had not received chemotherapy or radiotherapy before surgery.The clinical and pathological data such as clinical stage,tumor size,lymph node metastasis,and tissue differentiation of cervical cancer were recorded.Western blotting was used to study CCND1 and Wnt at the clinical tissue level./ ?-catenin pathway related molecules expression in cervical cancer;HeLa cervical cancer cell line was selected and cultured in a DMEM medium containing 10% calf serum in an incubator containing 5% CO2 and 37 ° C saturated humidity,0.25 % Trypsin digestion and passage,from psychiatry,nucleic acid and protein levels to detect the mechanism of baicalein on cervical cancer cells.CO-IP method was used to detect the relationship between related molecules.Mice were selected from the experimental animal room of the hospital.Fifty mice,all female.Select cell lines that are most sensitive to baicalein in cell experiments.Each mouse was subcutaneously inoculated with 2.5 × 106 live cervical cancer cells under the two hind paw pads and the back.After successful tumor bearing,the model mice were randomly divided into 4 groups according to the intervention method(8 in each group): control Group: Intratumoral injection of 0.1ml NS was the negative control group;Baicalin group: Intratumoral injection of baicalein was the positive control group,Low-dose group: Intratumoral injection of baicalein 20 mg / kg;Medium-dose group: Intratumoral injection of baicalein 40 mg / kg;baicalein high dose group: baicalein was given 80 mg / kg intratumorally.The tumor formation and general growth before and after the experiment were observed in each group of mice.The tumor size and the number of lymph node metastases were measured by vernier calipers before and on the 15 th days after treatment.Mice were sacrificed after 15 days of treatment.The tumors were stripped and weighed.The tumor suppression rate was calculated according to the following formula.CCND1 and Wnt / ?-catenin pathway-related proteins ?-catenin,Dsh,APC,GSK3,and MMP9 were detected by Western blot and fluorescent quantitative PCR.expression.Results:(1)In terms of CCND1,compared with cervical cancer tissues,long transcript transcription was relatively low,but there was no statistical difference(P> 0.05).The expression of short transcript transcription in cervical cancer tissues Significantly reduced,compared with adjacent tissues,P<0.05,the difference was statistically significant.The multiples of short transcript transcripts relative to long transcript transcripts were significantly different between groups(P<0.05).In terms of ?-catenin,the abnormal expression rate of ?-catenin in cervical cancer tissues was 78.00%,and the abnormal expression rate in adjacent tissues was 34.78%(P<0.05).In terms of Dsh,there was no statistical difference in the expression of Dsh in cervical cancer tissues and adjacent tissues(P> 0.05).In terms of APC,the negative and weak expression rates in cervical cancer tissues were significantly higher than those in adjacent cancer tissues,and the normal expression rates were significantly lower than those in adjacent cancer tissues(P<0.05).In terms of GSK3,the expression of GSK3 in cervical cancer tissues was significantly lower than that in adjacent tissues(P<0.05).In terms of MMP9,the negative and weak positive rates in cervical cancer tissues were significantly lower than those in adjacent tissues,and the strong positive expression rates were significantly higher in cervical tissues(P<0.05).(2)There are significant differences in the migration ability of HeLa cells in different groups,and the application of baicalein and the inhibition of the Wnt / ?-catenin pathway can effectively inhibit the migration ability of HeLa cells.At the same time,the combined application has an inhibitory effect on the migration ability of HeLa cells.Better.The invasion ability of HeLa cells in different groups is significantly different,and the application of baicalein and inhibition of the Wnt / ?-catenin pathway can effectively inhibit the invasion ability of HeLa cells.At the same time,the combined application has a better inhibitory effect on the invasion ability of HeLa cells.The administration of baicalein in HeLa cells combined with signal pathway inhibitory intervention,changes in CCND1 and Wnt / ?-catenin pathway related proteins(?-catenin,Dsh,APC,GSK3,MMP9)are shown in Figure 3.3 below.After intervention,CCND1,Dsh,and MMP9 expressions were down-regulated,APC and GSK3 expressions were upregulated,and ?-catenin was phosphorylated and degraded in the cytoplasm,which caused the inactivation of the Wnt / ?-catenin signaling pathway,which caused the biological activity of cervical cancer cells.inhibition.(3)In terms of tumor growth volume,there was a statistically significant difference between the control group and the baicalein group(P<0.05),and the tumor growth of cervical cancer mice had a significant dose-dependent relationship(P<0.05).In terms of the number of lymph node metastases,there was a statistically significant difference between the control group and the baicalein group(P<0.05),and the number of lymph node metastases in cervical cancer mice was significantly dose-dependent(P<0.05).In terms of tumor suppression rate,compared with the control group,the tumor weight of the baicalein high-dose and middle-dose groups was significantly reduced,and the tumor suppression rate was significantly increased.Moreover,there was a significant dose dependence.After inspection,P<0.05.After baicalein intervention,CCND1 and Wnt / ?-catenin pathway-related proteins(?-catenin,Dsh,APC,GSK3,MMP9)were significantly improved.Comparison between groups showed that the medium-dose and high-dose groups were significantly better than the control.The middle-dose and high-dose groups were superior to the baicalein group in terms of ?-catenin,Dsh,APC,GSK3,and MMP9.After testing,the difference was statistically significant at P<0.05.Conclusions:(1)Compared with adjacent cancer tissues,the expression of CCND1 in cervical cancer tissues is reduced,and the abnormal expression rate of ?-catenin in cervical cancer tissues is increased.The negative and weak expression rates in cervical cancer tissues are significantly increased.GSK3 expression was significantly reduced,negative expression and weak positive rate were significantly reduced in cervical cancer tissues,and there was no significant difference in Dsh expression.(2)The application of baicalein can inhibit the Wnt / ?-catenin pathway and further control the progression and metastasis of cervical cancer.CCND1,Dsh,MMP9 expression is down-regulated,APC and GSK3 expressions are up-regulated,and ?-catenin is in the cytoplasm.Phosphorylation and degradation cause inactivation of the Wnt / ?-catenin signaling pathway,which inhibits the biological activity of cervical cancer cells.(3)The application of baicalein can effectively control tumor growth volume,reduce lymph node metastasis,reduce tumor weight,and increase tumor suppression rate,making CCND1 and Wnt / ?-catenin pathwayrelated proteins(?-catenin,Dsh,APC,GSK3,MMP9)were significantly improved,and there was a significant dose dependence. |
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