| Prostate cancer is a common clinical malignant tumor in male.Its morbidity is related to age and androgen levels.With the improvement of the living standards and the aging of the population,the morbidity of prostate cancer in our country has continued to increase.Prostate cancer patients are prone to metastasis and recurrence,which is the main cause of death.Therefore,exploring the mechanism of prostate cancer metastasis is of great significance for the prevention and treatment of prostate cancer.Nonmuscle Myosin?(NM?)consists of two Nonmuscle myosin heavy chain(NMHC),two basic light chains(ELC),and two regulatory light chains(RLC).NMHC can bind to actin and directly regulate cell movement through autophosphorylation.NMHC? has three isoforms,namely NMHC?A,NMHC?B,and NMHC?C.NMHC?A plays an important role in immune response,cell migration,adhesion,and tumor metastasis.However,it is unclear whether NMHCIIA can affect metastasis in prostate cancer.Long noncoding RNA(lncRNA)is a type of noncoding RNA with more than 200 nucleotides.It has been found that abnormal expression of lncRNA plays a role in various diseases,especially in the invasion and metastasis of tumors,mainly through regulation of epigenetic,transcription,post-transcription,and miRNAs.It has been reported that LncRNA-HULC acts as a molecular sponge of miR-9-5p and regulates the miR-9-5p / MYH9 axis to promote gastric cancer progression.In papillary thyroid carcinoma,NMHCIIA inhibits the common promoter of FOXE1 and PTCSC2 by binding to LncRNA-PTCSC2,thereby inhibiting the p53 pathway and promoting papillary thyroid carcinoma progression.But it is unclear whether there is a relationship of regulation between lncRNA and NMHCIIA in prostate cancer.Hence,we try to explore whether the lncRNA can regulate NMHCIIA in prostate cancer,and then affect the metastasis of prostate cancer.Objective:To investigate the role of NMHC?A in prostate cancer metastasis and to screen the regulatory axis of LncRNA related to NMHC?A in prostate cancer.Methods:1.We used the Oncomine database to analyze the mRNA expression level of MYH9 in prostate cancer.Western bolt was used to detect the NMHCIIA in the benign prostatic hyperplasia cell line BPH-1,the prostate cancer cell line PC3,and the high metastatic prostate cancer cell line PC3 M.2.We used the technique of RNA interference to silence NMHC?A gene PC3 M cell,and selected out the stable cell lines.The efficiency of silence was detected via western blot and Immunofluorescence.3.Through experiments in vitro studey the effect of gene silence of NMHC?A on biological behaviors of PC3 M cell.The CCK-8 kit was used to detect cell proliferation,the Transwell migration tests was used to detect cell migration,and the Transwell invasion tests was used to detect cell invasion.4.R is used to screen the transcriptome data of prostate cancer patients in the database of The Cancer Genome Atlas(TCGA).The abnormal expression of lncRNA and miRNA in prostate cancer patients were obtained and matched.The miRcode,miRDB,and Targetscan databases predict miRNAs targeting MYH9 online,compare the online prediction results with the TCGA screening results,obtain common miRNAs,and find their upstream lncRNA,and then speculate the lncRNA / miRNA / MYH9 axis.Results:1.Expression of NMHC?A in prostate cancer.1.1 Oncomine database analysis showed that there were 452 unique analyses for MYH9 expression in different types of tumors and normal tissues,and 49 studies showed that a significant statistical difference for MYH9(P <0.01).In the ‘Wallace Prostate' dataset,the expression of MYH9 in prostate cancer is higher than normal(P = 2.29E-4).1.2 Compared with BPH-1 cells,the expression of NMHCIIA was higher in PC3 and PC3 M cells(P<0.05).And compared with PC3 cells,NMHCIIA expression was increased in PC3 M cells(P<0.05).2.Under the fluorescence microscope,the infection efficiency of each group was over 80%.The results of western bolt and immunofluorescence showed that there was no significant difference in the expression of NMHCIIA between the control group and the control vector group(P>0.05);the expression level of NMHC?A in the shMYH9 group was significantly lower than non-transfection group and control vector group(P<0.05).3.Effect of gene silence of NMHC?A on biological behaviors in prostate cancer cell line PC3 M.3.1 CCK-8 kit test results showed that there was no significant difference of the proliferation between the control group and the control vector group(P>0.05),and the proliferation of the shMYH9 group was significantly lower than the control group and the control vector group(P<0.05).3.2 Transwell migration tests results t showed that there was no significant difference in the number of cells that passed through the basement membrane between the control group and the control vector group(P>0.05),while the cells that passed through the basement membrane of the shMYH9 group was significantly less than the control group and the control vector group(P<0.05).3.3 Transwell invasion tests results t showed that there was no significant difference in the number of cells passing through the basement membrane which degraded Matrigel between the control group and the control vector group(P>0.05),while the cells of the shMYH9 group was significantly less than the control group and the control vector group(P<0.05).3.4 The results of TCGA database screening and miRcode,miRDB,and Targetscan database online prediction showed that KIAA0087/mir-133b/ MYH9,HNF1A-AS1/mir-133b/ MYH9,CACNA1C-IT3/mir-133b/ MYH9,three axes may play a role in regulating NMHCIIA in prostate cancer.Conclusions:1.The high expression of NMHCIIA may be related to prostate cancer metastasis.2.NMHC?A may regulate the metastasis of prostate cancer by regulating the migration,invasion and proliferation of prostate cancer cells.3.LncRNA may regulate NMHCIIA through KIAA0087 / mir-133 b / MYH9,HNF1A-AS1 / mir-133 b / MYH9,CACNA1C-IT3 / mir-133 b / MYH9,three potential axes in prostate cancer. |
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