长链非编码RNA-PVT1促进肾癌细胞的迁移和转移

Long non-coding RNA-PVT1 promotes migration and metastasis of renal carcinoma cells

目的探讨长链非编码RNA-PVT1(Lnc-PVT1)在透明细胞肾细胞癌(ccRCC)中的表达水平及临床意义。方法收集69对ccRCC组织和癌旁正常组织。采用实时荧光定量聚合酶链反应(qRT-PCR)检测Lnc-PVT1在ccRCC中的表达水平。应用RNA原位杂交(RISH)技术测定ccRCC中Lnc-PVT1阳性表达情况。采用Transwell实验分析ccRCC细胞迁移的数目变化。通过生物信息学预测Lnc-PVT1和微小RNA-145(miR-145)的下游靶基因,并通过实验加以验证。采用Rescue回复实验分析Lnc-PVT1对miR-145/基质金属蛋白酶-9(MMP9)通路和迁移的影响。结果qRT-PCR结果显示,Lnc-PVT1在ccRCC组织和细胞中表达水平升高(P<0.05)。RISH实验提示,Lnc-PVT1在ccRCC组织中呈阳性表达。ccRCC组织Lnc-PVT1阳性率为73.91%(51/69),高于癌旁正常组织的14.49%(10/69),差异有统计学意义(χ^(2)=4.128,P<0.05)。Lnc-PVT1的阳性表达与ccRCC病理分级和淋巴结转移相关(P<0.05),而与患者年龄、性别、肿瘤大小和TNM分期无关(P>0.05)。Transwell迁移实验结果表明,过表达Lnc-PVT1促进ccRCC细胞迁移能力,降低Lnc-PVT1表达抑制ccRCC细胞迁移能力(P<0.05)。miR-145是Lnc-PVT1调控的靶基因;miR-145下游直接靶基因是MMP9。Lnc-PVT1能抑制miR-145表达,促进MMP9蛋白表达。结论Lnc-PVT1通过miR-145/MMP9通路促进肾癌细胞的迁移和转移,或可为治疗转移性肾细胞癌提供可能的分子靶点和依据。

Objective To investigate the expression level and clinical significance of long non-coding RNA-PVT1(Lnc-PVT1)in clear cell renal cell carcinoma(ccRCC).Methods Sixty-nine pairs of ccRCC tissues and adjacent normal tissues were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression level of Lnc-PVT1 in ccRCC.RNA in situ hybridization(RISH)was used to determine the positive expression of Lnc-PVT1 in ccRCC.Transwell assay was used to analyze the number of cell migration in ccRCC.The downstream target genes of Lnc-PVT1 and microRNA-145(miR-145)were predicted by bioinformatics and were verified by experiments.The effects of Lnc-PVT1 on miR-145/matrix metalloproteinase-9(MMP9)pathway and migration were analyzed by Rescue recovery experiment.Results The results of qRT-PCR showed that the expression level of Lnc-PVT1 in ccRCC tissues and cells was increased(P<0.05).RISH assay indicated that Lnc-PVT1 was positively expressed in ccRCC tissue.The positive rate of Lnc-PVT11 in ccRCC tissue was 73.91%(51/69),which was higher than 14.49%of adjacent normal tissues(10/69),the difference was statistically significant(χ^(2)=4.128,P<0.05).The positive expression of Lnc-PVT1 was correlated with ccRCC pathological grade and lymph node metastasis(P<0.05),but had no correlations with patient age,sex,tumor size and TNM stage(P>0.05).The results of Transwell migration experiment showed that overexpression of Lnc-PVT1 promoted the migration ability of ccRCC cells,and decreased expression of Lnc-PVT1 inhibited the migration ability of ccRCC cells(P<0.05).The miR-145 was a target gene regulated by Lnc-PVT1;the downstream direct target gene of miR-145 was MMP9.Lnc-PVT1 could inhibit the expression of miR-145 and promote the expression of MMP9 protein.Conclusion Lnc-PVT1 promotes migration and metastasis of renal cancer cells through the miR-145/MMP9 pathway.This study may provide a possible molecular target and basis for the treatment of metastatic renal cell carcinoma.