The Function And Mechanism Of Regulation Of MiRNA-21Targeted PDCD4in Prostate-cancer Progresse

Objective:Respectively.to predict and verify the regulation of miR-21targeting the PDCD4by molecular biology and bioinformatics in prostate cancer cell line PC-3and LNCap.To investigate the relationship between miR-21and PDCD4during prostate cancer development and progression..Our study will propose new methods and experiment dates for diagnosis and treatment of prostate cancer,especially androgen-independent prostate cancer.Methods:1, Transfection by lipofectamine and qRT-PCR:To detect the relative expression level of PDCD4before and after transfection,by transiently transfected with anti-miR-21and anti-miR-21negative control performed with lipofectamine into prostate cancer cell line PC-3and LNCap.Extracted total RNA from prostate cancer cell line PC-3and LNCap adding to anti-miR-21and anti-miR Negative control or not,and then detected expression of PDCD4at the transcription level.2, Western blot: Extracted total protin from prostate cancer cell line PC-3and LNCap adding to anti-miR-21and anti-miR Negative control or not,after purification,electrophoresis,transmembrane,incubated antibody of PDCD4,film exposuring and developing, detected expression of PDCD4at the translation level.3, Immunohistochemistry:To analyze the expression differences of PDCD4and PSA in protin level in prostate cancer tissue, border district tissue and normal prostate tissue (>15cases).Reselts: 1, The result for qRT-PCR:After the anti-miR-21inhibitors and its negative controls were respectively transfected into PC-3and LNCap, the expression level of PDCD4of both cell lines in transcription was down-regulated compared with negative control. Respectively,the z-value were-3.111and-3.180,obtaining by statistical analysis,and the bilateral probability were0.002and0.001.At the a level of0.05, Ho must be abandoned.2, The result for Western Blot:After transfection,to a certain extent,the expression of PDCD4in both cell lines in translation level was decreased,but that of its negative control did not change obviously compared with the blank group.After statistical analysis,the z-value were respectively-3.920and-3.883,and both of the bilateral probability belew0.001, Ho was discarded at the a level of0.05.The result suggested a significantly difference.After negative control was transfected,the expression of PDCD4protein did not change significantly compared with the blank group,and the z-value were-1.643and-0.971,the bilateral probability0.100and0.332respectively.No statistically significant difference during the two sets of data at the a level of0.05.3, The result of immunohistochemistry showed the expression of the PDCD4in protate cancer,precancerous lesions and prostatic hyperplasia tissue is up-regulated in order,and the that of the PSA showed a negative correlation trend comparing with PDCD4.By Chi-square test,the outcome (x2=8.632, P＜.05) implied that the expression of PDCD4and PSA did not presenced obvious diversity in different tissue.It showed an opposite expression trend between PDCD4and PSA by Spearman test (-1<r<0)Conclusions:1, MiRNA-21regulate the expression of the tumor-suprpressor gene PDCD4in prostate cancer,PDCD4is one of the targeting regulatory gene of miR-21.2,The expression level of PDCD4is difference in different prostate tissue,so it will be a promising tumor markers for both of the diagnosisi of prostate cancer and its follow-up.3, The miR-21targeting PDCD4tuning tips:This regulatory approach may be a new therapeutic target for prostate cancer,especially for androgen-independent prostate cancer.