| Background Docetaxel is the first- line chemotherapy drugs for prostate cancer, and its curative effect needs to be improved. Nobiletin could inhibit the proliferation of tumor cells. DAB2 IP is a tumor suppressor gene, which lost expression in the majority of tumors, and its high expression could inhibit the proliferation and metastasis of tumor cells. EZH2 was highly expressed in some aggressive or metastatic tumors, which suggested that the EZH2 expression is higher, the higher the degree of malignancy, the stronger aggressive. It was found that EZH2 could cut the expression of DAB2 IP, which was involved in tumor progression. Previous studies had successfully transfected DAB2 IP gene into PC3 cells. Aiming at providing a new method for the treatment of prostate cancer and targets, this research intends to study nobiletin in combination with docetaxel, which discusses the proliferation, apoptosis and migration ability in prostate cancer PC3 and DAB2IP+/PC3 cell, and investigate the difference and clinical significance of DAB2 IP and EZH2 expression in prostate diseases.PART? A study about the effect of nobiletin and docetaxel on the growth of prostate cancer PC3 and DAB2IP+/PC3 cellsObjective To explore the effect of nobiletin and docetaxel on prostate cancer PC3 and DAB2IP+/PC3 cell proliferation, apoptosis and migration.Methods MTS was applied to detect the proliferation in PC3 and DAB2IP+/PC3 cells by nobiletin(concentration is 6.25, 12.5, 25, 50, 62.5?mol/L), docetaxel(concentration is 10-5, 10-6, 10-7mol/L) and combined medication after 24 and 48 hours, and set four negative holes in each concentration groups, and also set the blank group with no drug. Scarification test was applied to detect the ability of migration in PC3 and DAB2IP+/PC3 cells by nobiletin(concentration is 62.5?mol/L) and docetaxel(concentration is 10-5, 10-7mol/L) and combined medication after 24 and 48 hours, and also set the blank group with no drug. Flow cytometry was used to detect the effect of the apoptosis by nobiletin(concentration is 62.5?mol/L) and docetaxel(concentration is 10-5, 10-7mol/L) and combined medication after 24 and 48 hours, and also set the blank group with no drug.Results The MTS results: after nobiletin affect 24 and 48 hours, the same concentration of nobiletin in transfected group compared with empty plasmid group, the differences are statistically significant(P<0.05). When the different concentration of nobiletin in empty plasmid group, blank group and 6.25?mol/L group compared with 62.5?mol/L group, the differences are statistically significant(P<0.05). When the different concentrations of nobiletin in transfected group, blank group, 6.25?mol/L group and 12.5?mol/L group compared with 62.5?mol/L group, the differences are statistically significant(P<0.05). And 6.25?mol/L group compared with 50?mol/L group, the difference is statistically significant(P<0.05). After docetaxel affect 24 and 48 hours, the same concentration of docetaxel in transfected group compared with empty plasmid group, the differences are statistically significant(P<0.05). Docetaxel(10-5, 10-6, 10-7mol/L)group compared with blank group, the differences are statistically significant(P<0.05). The differences between different concentrations of docetaxel are statistically significant(P>0.05). After nobiletin(concentration is 6.25, 12.5, 25, 50, 62.5?mol/L) combine with docetaxel(concentration is 10-5, 10-6, 10-7mol/L) affect 24 and 48 hours, effection of the same combined concentration in transfected group compared with empty plasmid group, the difference is statistically significant(P<0.05).(1)Empty plasmid group: when the same concentration of docetaxel(10-5mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group compared with 6.25?mol/L group, the difference is statistically significant(P<0.05). When the same concentration of docetaxel(10-6mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group compared with 6.25?mol/L group, the difference is statistically significant(P<0.05). When the same concentration of docetaxel(10-7mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group and 50?mol/L group compared with 6.25?mol/L group, and 62.5?mol/L group compared with 12.5?mol/L group, the differences are statistically significant(P<0.05). When the sameconcentration of nobiletin(62.5?mol/L) combine with the different concentrations of docetaxel, 10-5mol/L group compared with 10-7mol/L group, the difference is statistically significant(P<0.05). The other groups were not statistically significant.(2)Transfected group: when the same concentration of docetaxel(10-5mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group and 50?mol/L group compared with 6.25?mol/L group, and 62.5?mol/L group compared with 12.5?mol/L group, the differences are statistically significant(P<0.05). When the same concentration of docetaxel(10-6mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group and 50?mol/L group compared with 6.25?mol/L group, 62.5?mol/L group and 50?mol/L group compared with 12.5?mol/L group, the differences are statistically significant(P<0.05). When the same concentration of docetaxel(10-7mol/L) combine with the different concentrations of nobiletin, 62.5?mol/L group, 50?mol/L group and 25?mol/L compared with 6.25?mol/L group, and 62.5?mol/L group and 50?mol/L group compared with 12.5?mol/L group, the differences are statistically significant(P<0.05). When the same concentration of nobiletin(62.5?mol/L) combine with the different concentrations of docetaxel, 10-5mol/L group compared with 10-7mol/L group, the difference is statistically significant(P<0.05). The other groups were not statistically significant. The migration results: after different drugs affect 24 and 48 hours, the same concentration of drug in transfected group compared with empty plasmid group, the differences are statistically significant(P<0.05). Two groups of cells' migration rate in each group compared with blank group, the differences are statistically significant(P<0.05). Docetaxel 10-5mol/L group compared with 10-7mol/L group, the difference is statistically significant(P<0.05). Docetaxel(10-5mol/L, 10-7mol/L) groups compared with nobiletin group, the differences are statistically significant(P<0.05). Combined group compared with nobiletin group, the differences are statistically significant(P<0.05). Docetaxel 10-5mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-5mol/L group, the differences are statistically significant(P<0.05). Docetaxel 10-7mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-7mol/L group, the differences are statistically significant(P<0.05).Docetaxel 10-5mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-7mol/L and nobiletin 62.5?mol/L group, the differences are statistically significant(P<0.05). The apoptosis results: after different drugs affect 24 and 48 hours, the same concentration of drug in transfected group compared with empty plasmid group, the differences are statistically significant(P<0.05). Two groups of cells' apoptosis rate in each group compared with blank group, the differences are statistically significant(P<0.05). Docetaxel 10-5mol/L group compared with 10-7mol/L group, the difference is statistically significant(P<0.05). Docetaxel(10-5mol/L, 10-7mol/L) groups compared with nobiletin group, the differences are statistically significant(P<0.05). Combined group compared with nobiletin group, the differences are statistically significant(P<0.05). Docetaxel 10-5mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-5mol/L group, the differences are statistically significant(P<0.05). Docetaxel 10-7mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-7mol/L group, the differences are statistically significant(P<0.05). Docetaxel 10-5mol/L and nobiletin 62.5?mol/L group compared with Docetaxel 10-7mol/L and nobiletin 62.5?mol/L group, the differences are statistically significant(P<0.05).Conclusion 1.Nobiletin and docetaxel inhibit the proliferation of prostate cancer PC3 cell. 2.The effect of nobiletin combined with docetaxel is better than single drug, and nobiletin can reduce the concentration of docetaxel, which shows that nobiletin can improve the effect of chemotherapeutic drugs. 3.DAB2 IP transfected can inhibit the proliferation of prostate cancer PC3 cell. DAB2 IP transfected can improve the effect of drugs.PART ? A study of the expression of DAB2 IP and EZH2 in prostate diseaseObjective To investigate the expression and clinical significance of DAB2 IP and EZH2 in prostate cancer and benign prostatic hyperplasia.Methods Randomly selected 40 cases of prostate ca ncer and 10 cases of benign prostatic hyperplasia patients, all cases of prostate cancer have no endocrine therapy, radiotherapy or chemotherapy. Remove the specimens from pathologic specimens library, each tissue wax was cut into two pieces for detecting the DAB2 IP and EZH2 by immunohistochemical method.Results EZH2 is mainly expressed in the nucleus, which is pale brown. DAB2 IP is mainly expressed in the cytoplasm, which is pale brown or dark brown. The positive rate of EZH2 expression in prostate cancer tissues(77.5%) was higher than that in benign prostatic hyperplasia tissues(30%), and the difference is statistically significant(P<0.05). The positive rate of DAB2 IP expression in benign prostatic hyperplasia tissues(80%) is higher than that in prostate cancer tissues(27.5%), and the difference is statistically significant(P<0.05). There is a negative correlation between EZH2 and DAB2 IP expression in prostate cancer and benign prostatic hyperplasia(P<0.05). There is a positive correlation between the EZH2 expression and Gleason score in prostate cancer(P<0.05).Conclusion 1. EZH2 is high expressed in prostate cancer and low expressed in benign prostatic hyperplasia. 2. The expression of EZH2 is positively correlated with Gleason score in prostate cancer. The expression of EZH2 is negatively correlated with the expression of DAB2 IP in prostate cancer tissue. |
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