Abstract
Background Bladder cancer (BC) is a malignant and common malignant tumors. However, the prognosis of most patients with bladder cancer is still poor, and it is particularly important to identify early tumor diagnostic and treatment targets. Materials and Methods High-throughput sequencing was used to evaluate the expression level of circRNA in bladder cancer tissue. MTT assay, wound healing assay, and transwell assay were used to detect the cancer cells' proliferation, migration, and invasion affected by hsa_circ_0139402. The possible miRNA targets of hsa_circ_0139402 and downstream genes were detected by bioinformatics methods and dual-luciferase reporting experiment. FISH was used to observe their interaction. Results High-throughput sequencing result showed that the expression of hsa_circ_0139402 was highest in BC tissues and increased in metastatic tissues compared to that of nonmetastatic tissues. MTT assay, wound healing assay, and transwell assay revealed that sh-hsa_circ_0139402 could suppress BC cells' proliferation, invasion, and migration. Bioinformatics analysis, dual-luciferase reporter, and RIP assay showed that hsa_circ_0139402 can bind to hsa-miR-326, and PAX8 is a direct target of hsa-miR-326 in BC cell. Further, cytological studies found that hsa_circ_0139402 enhances BC cells' proliferation, migration, and invasion by targeting PAX8 via hsa-miR-326. Conclusion hsa_circ_0139402 plays a oncogene in BC and that can effectively promote cell proliferation, migration, invasion, and EMT by targeting Paired Box Protein Pax-8 (PAX8) via hsa-miR-326 and provides a potential therapeutic target for BC patients.
Highlights
Bladder cancer (BC) is a malignant tumor that occurs on the mucous membrane of the bladder and is one of the most common malignant tumors
The cell lines were identified by the Short Tandem Repeat (STR) method (Biowing, China). shRNA and overexpression of hsa_circ_0139402 were obtained from GenePharma company (Shanghai, China). hsa-miR-326 mimics were purchased from RiboBio (Guangzhou, China), and cell transfection was performed using Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer’s protocol
High-throughput sequencing results showed that 295 Circular RNA (circRNA) has abnormal expression in BC tissues, and the expression of hsa_circ_0139402 was highest in BC tissues (Figure 1(a)); RT-PCR result confirmed that the expression of hsa_circ_0139402 was upregulated in 39 pairs of BC tissues and increased in metastatic tissues compared to that of nonmetastatic tissues (32 pairs of BC tissues) (Figures 1(b) and 1(c) and Table 1, ∗P < 0:05); hsa_circ_ 0139402 expression was increased in UMUC3 and 5637
Summary
Bladder cancer (BC) is a malignant and common malignant tumors. the prognosis of most patients with bladder cancer is still poor, and it is important to identify early tumor diagnostic and treatment targets. High-throughput sequencing was used to evaluate the expression level of circRNA in bladder cancer tissue. MTT assay, wound healing assay, and transwell assay were used to detect the cancer cells’ proliferation, migration, and invasion affected by hsa_circ_0139402. The possible miRNA targets of hsa_circ_0139402 and downstream genes were detected by bioinformatics methods and dual-luciferase reporting experiment. High-throughput sequencing result showed that the expression of hsa_circ_0139402 was highest in BC tissues and increased in metastatic tissues compared to that of nonmetastatic tissues. MTT assay, wound healing assay, and transwell assay revealed that sh-hsa_circ_ 0139402 could suppress BC cells’ proliferation, invasion, and migration. Bioinformatics analysis, dual-luciferase reporter, and RIP assay showed that hsa_circ_0139402 can bind to hsa-miR-326, and PAX8 is a direct target of hsa-miR-326 in BC cell. Cytological studies found that hsa_circ_0139402 enhances BC cells’ proliferation, migration, and invasion by targeting PAX8 via hsa-miR-326. Conclusion. hsa_circ_0139402 plays a oncogene in BC and that can effectively promote cell proliferation, migration, invasion, and EMT by targeting Paired Box Protein Pax-8 (PAX8) via hsa-miR-326 and provides a potential therapeutic target for BC patients
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