LncRNA DSCAM-AS1通过miR-144-5p激活PI3K-AKT通路对前列腺癌细胞的影响

Effects of LncRNA DSCAM-AS1 on prostate cancer cells by activating the PI3K-AKT pathway through miR-144-5p

目的探讨LncRNA DSCAM-AS1通过miR-144-5p激活PI3K-AKT通路对前列腺癌细胞的影响机制。方法选取2019年8月至2020年12月新疆医科大学第二附属医院确诊的前列腺癌患者的50例癌组织标本和38例癌旁正常组织标本,以及前列腺癌PC-2、DU145细胞和人正常前列腺上皮RWPE-1细胞作为研究对象。采用实时荧光定量聚合酶链反应(qRT-PCR)检测LncRNA DSCAM-AS1和miR-144-5p的表达;分别下调或上调DSCAM-AS1与miR-144-5p的表达后,采用MTT实验检测DU145细胞活力、Transwell检测DU145细胞侵袭数目及Western blot检测DU145细胞上皮间质转化(EMT)相关蛋白表达;采用miRcode预测LncRNA DSCAM-AS1的靶基因;上调或下调DSCAM-AS1表达后,采用Western blot检测p-PI3K、PI3K、p-AKT和AKT的蛋白表达;将LY294002作用于DU145细胞,检测DU145细胞增殖、侵袭和EMT情况。结果前列腺癌组织和癌旁正常组织中DSCAM-AS1表达分别为2.28±0.42和1.06±0.71,miR-144-5p表达分别为0.37±0.24和1.19±0.96,二者比较,差异具有统计学意义(P<0.01);DU145和RWPE-1细胞中DSCAM-AS1表达分别为3.45±1.28和1.15±0.92,miR-144-5p表达分别为0.23±0.16和1.18±0.63,二者比较,差异具有统计学意义(P<0.01)。下调LncRNA DSCAM-AS1会抑制DU145细胞的活力、侵袭和EMT;LncRNA DSCAM-AS1靶向调控miR-144-5p;上调LncRNA DSCAM-AS1通过miR-144-5p促进DU145细胞的增殖、侵袭和EMT。将pcDNA-DSCAM-AS1和LY294002作用于DU145细胞会明显抑制DU145细胞的活力、侵袭和EMT。结论上调LncRNA DSCAM-AS1会通过miR-144-5p激活PI3K-AKT信号通路促进前列腺癌细胞的发展。

Objective To investigate the effect of LncRNA DSCAM-AS1 on the development of prostate cancer cells by activating the PI3 K-AKT pathway through miR-144-5 p.Methods 50 cancer tissue samples and 38 normal adjacent tissue samples from prostate cancer patients diagnosed in the Second Affiliated Hospital of Xinjiang Medical University from August 2019 to December 2020,as well as prostate cancer PC-2 cells,DU145 cells and human normal prostate epithelial RWPE-1 cells were selected as the research objects.The expression of LncRNA DSCAM-AS1 and miR-144-5 p was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).After down-regulating or up-regulating the expressions of DSCAM-AS1 and miR-144-5 p,DU145 cell viability was detected by MTT assay,DU145 cell invasion number was detected by Transwell,and EMT-related protein expression was detected by Western blot.The target gene of LncRNA DSCAM-AS1 was predicted by miRcode.After up-regulation or down-regulation of DSCAM-AS1 expression,the protein expressions of p-PI3 K,PI3 K,p-AKT and AKT were detected by Western blot.LY294002 was applied to DU145 cells to detect the proliferation,invasion and EMT of DU145 cells.Results The expression levels of DSCAM-AS1 in prostate cancer tissues and adjacent normal tissues were 2.28±0.42 and 1.06±0.71,and the expression levels of miR-144-5 p were 0.37±0.24 and 1.19±0.96,which were statistically significant(P<0.01).The expression levels of DSCAM-AS1 in DU145 and RWPE-1 cells were 3.45±1.28 and 1.15±0.92,respectively,and the expression levels of miR-144-5 p were 0.23±0.16 and 1.18±0.63,respectively,which were statistically significant(P<0.01).Down-regulation of LncRNA DSCAM-AS1 inhibited the activity,invasion and EMT of DU145 cells.LncRNA DSCAM-AS1 targeted miR-144-5 p.Up-regulating LncRNA DSCAM-AS1 promoted proliferation,invasion and EMT of DU145 cells through miR-144-5 p.PcDNA-DSCAM-AS1 and LY294002 significantly inhibited the the activity,invasion and EMT of DU145 cells.Conclusions Up-regulation of LncRNA DSCAM-AS1 can activate the PI3 K-AKT signaling pathway through miR-144-5 p to promote the development of prostate cancer cells.