长链非编码RNA PVT1抑制miR-31表达并促进卵巢癌细胞侵袭转移的实验研究

Long non-coding RNA PVT1 promoted metastasis via suppression of miR-31 in ovarian cancer cells

目的:探讨长链非编码RNA PVT1能否抑制卵巢癌细胞侵袭转移,明确其影响可否通过抑制miR-31的方式实现。方法:qRT-PCR法检测长链非编码RNA PVT1及miR-31在卵巢癌组织及卵巢癌细胞系SKOV3中的表达,同时分别设定正常卵巢组织和卵巢正常上皮细胞系IOSE80作为对照。研究PVT1表达水平与卵巢癌患者临床病理参数的相关性。SKOV3细胞中转染PVT1 siRNA下调PVT1表达,qRT-PCR检测PVT1表达对miR-31表达的影响,Transwell实验检测PVT1表达对SKOV3侵袭转移能力的影响。“反义”实验验证下调miR-31能否逆转PVT1 siRNA对SKOV3侵袭转移能力的抑制效应。荧光素酶报告基因实验检测miR-31与PVT1的靶向结合作用。结果:卵巢癌组织及细胞系中,PVT1表达显著增高,miR-31表达显著降低,两者表达呈明显负相关。PVT1高表达与卵巢癌患者的高临床分期及腹膜后转移密切相关。转染PVT1 siRNA可明显下调SKOV3中PVT1表达,促进miR-31表达,抑制SKOV3细胞的侵袭转移能力;转染miR-31 inhibitor可明显逆转PVT1 siRNA对SKOV3侵袭转移能力的抑制效应;荧光素酶报告基因实验证实miR-31可与PVT1靶向结合。结论:PVT1可通过抑制miR-31的方式促进卵巢癌细胞侵袭转移。

Objective:To investigate whether long noncoding RNA PVT1 could inhibit ovarian cancer(OC)cells metastasis and to explore whether the suppressive effect was achieved through inhibition of miR-31 pathway.Methods:The expression of long noncoding RNA PVT1(lncRNA PVT1)and microRNA-31(miR-31)in OC tissue and cell lines were measured by qRT-PCR.To detect the correlation between PVT1 and clinicopathological features of patients with OC.PVT1 in SKOV3 cells was knocked down by a transfection of PVT1 siRNA.The expression of miR-31 after PVT1 knockdown was determined by qRT-PCR.The metastatic ability change of SKOV3 cells was evaluated by a transwell assay.An anti-sense experiment was used to explore whether miR-31 could reverse the suppressive effect which PVT1 siRNA did on SKOV3 cells metastasis.A luciferase assay was applied to check the targeted binding effect between PVT1 and miR-31.Results:Upregulated PVT1 but downregulated miR-31 were expressed in OC.The expressions of PVT1 and miR-31 presented a markedly negative correlation.High expressed PVT1 was closely correlated with higher clinical staging and retroperitoneal metastasis in patients with OC.A transfection of PVT1 siRNA suppressed PVT1 and its mediated metastasis but promoted miR-31 expression in SKOV3 cells.A transfection of miR-31 inhibitor attenuated the inhibitive effect which PVT1 siRNA did on SKOV3 cells metastasis.The results of luciferase assay showed that miR-31 could target PVT1.Conclusions:PVT1 promotes OC cells metastasis via suppression of miR-31.