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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
lncRNA PAX8-AS1 suppresses cervical cancer development by regulating miR-675-3p/DCN axis
Xiayang Lu1, Peng Song2, Qingfen Chen3
1Gastroenterology Department, Taizhou Traditional Chinese Medicine Hospital, Zhejiang Province, China.
Abstract:
ObjectiveCervical cancer (CC) is among the most prevalent malignancies globally. Public sequencing data indicate that PAX8-AS1 is associated with gynecological cancers, including CC, but its specific function and mechanism in cervical cancer remain unclear. The present study aimed to elucidate the role of PAX8-AS1 and its target axis in the CC.MethodsA total of 104 CC patients were included. The levels of PAX8-AS1, miR-675-3p, and decorin (DCN) were quantified by quantitative reverse transcription polymerase chain reaction. Survival curve and Cox regression predicted prognostic factors. Proliferation and invasion/migration were assayed by CCK-8 and Transwell in CC cell lines. The target relationship was verified by dual-luciferase reporter assay and co-transfection.ResultsPAX8-AS1 was declined in CC tumor tissue and cell lines. PAX8-AS1 was an independent prognostic factor. PAX8-AS1 expression was associated with the CC pathological including the international federation of gynecology and obstetrics staging system (FIGO), tumor size, and lymph node metastasis. Patients with higher PAX8-AS1 levels had better survival outcomes. Upregulation of PAX8-AS1 inhibited the CC cell invasion, migration, and proliferation. miR-675-3p was predicted and verified as sponged of PAX8-AS1. miR-675-3p was negatively related to PAX8-AS1. PAX8-AS1 impeded the CC cellular function by regulating miR-675-5p. DCN was confirmed as the target of miR-675-5p. DCN was negatively and positively related to miR-675-3p and PAX8-AS1, respectively. PAX8-AS1 suppressed the CC cell invasion, migration, and proliferation by targeting miR-675-3p/DCN axis.ConclusionIn summary, PAX8-AS1 was related to tumor progression and inhibited CC development. As a potential biomarker, PAX8-AS1 impeded CC cell invasion, migration, and proliferation by regulating the axis of miR-675-3p/DCN.
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