The Study Of FilaminA On Evolvement Of Prostate Cancer

Objective:To study the change of Filamin A on the evolvement of prostate cancer,and analysis its role and significance on carcinogenesis and evolvement of prostate cancer.Methods:Using RNA interference(RNAi) technique, pSilencer-Filamin A was transfected into the human prostate cell line LNCap by lipofectamine-mediation to inhibit the expression of Filamin A gene.The effect of RNAi on the expression of Filamin A gene were identified by Western blot;Selecting for prostate cancer cell lines that stably express the Filamin A siRNA through hygromycin-resistant selection.the cell growth curve were measured through methods of MTT,the cytomorphological alterations of LNCap cells transfected with pSilencer-Filamin A were observed under light microscope,soft agar colony formation were employed to analyze the ability of anchorage-independent growth, scarification test were employed to analyze the ability of cell migration.The expression of Filamin A in benign prostatic hyperplasia tissues and prostate cancer tissues was detected by immunohistochemistry and was analyzed by microscopic scoring and computer picture -scanning analysis system.Results:1.The RNAi vector of Filamin A was transfected into the human prostate cell line LNCap through hygromycin-resistant selection.Western-blot showed that RNAi vector could effectively inhibit the expression of Filamin A;2.the cytomorphological alterations:Morphology showed that the human prostate cell line LNCap arranged intensively,muscular,the number of adherence tentacle is more.The cell transfected with pSilencer-Filamin A showed sparsely distribution,shape was fibriform,the number of adherence tentacle is few.3.The cell growth curve was measured by MTT methods.The result showed that the growth rate of the human prostate cell line LNCap was much more accelerated than the cell transfected with pSilencer-Filamin A,compared with the growth rates of the two cells(p<0.05).4.The proliferative capacity of two group cells in soft agar was assessed by soft agar colony formation.The result indicated that the ability of the cell transfected with pSilencer-Filamin A to grow in soft agar is obviously stronger than the human prostate cell line LNCap,compared with the difference of the two cells(p<0.05).5.The cell migration capacity of two group cells was assessed by scarification test.The result indicated that the ability of the cell transfected with pSilencer-Filamin A to migration is obviously stronger than the human prostate cell line LNCap,compared with the difference of the two cells (p<0.05).6.The expression of Filamin A in benign prostatic hyperplasia tissues are higher than in prostate cancer tissues(expression ratio:27.3%,86.4 %,,P<0.0001,P<0.001).The expression of Filamin A is negative correlated with Gleason of prostate cancer.Conclusions:1.The RNAi vector of pSilencer-Filamin A could effectively inhibit the expression of Filamin A gene.2.The inhibition of Filamin A enhances the malignant level of prostate cancer cells.This indicated that the gene of Filamin A may be a anti-oncogene.3.The immunohistochemistry experiment in prostate cancer tissues showed that the expression of Filamin A gene is lower than in benign prostatic hyperplasia tissues.4.The expression of Filamin A is negative correlated with Gleason of prostate cancer.This indicated that the gene of Filamin A may be a marker as the grade of prostate cancer.In conclusion,the model of prostate cancer cell of inhibiting Filamin A were constructed successfully.Based on this study,the effect of Filamin A gene expression on the biological characteristics of the human prostate cell line LNCap was directly observed.The immunohistochemistry experiment in prostate cancer tissues and in benign prostatic hyperplasia tissues showed that the expression of Filamin A gene have a difference.The results indicated that Filamin A gene expression associated with prostate cancer.This work provides an important basis for further study of the biological functions of the Filamin A gene and its role in the occurrence and development of prostate cancer.