LncRNA KCNQ1OT1调节miR-145/PAK4轴对结直肠癌SW480细胞恶性发展的影响

doi:10.3969/j.issn.1006-6233.2023.02.02

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Abstract Objective: To explore the role of LncRNA KCNQ1OT1 in colorectal cancer SW480 cells and its potential molecular mechanism. Methods: MiRanda and TargetScan were conducted to analyze the action targets of LncRNA KCNQ1OT1 and miR-145 respectively. The intermolecular target relationship was verified by dual luciferase reporter assay. The qRT-PCR assay was used to analyze the gene expression levels of KCNQ1OT1, miR-145, and PAK4 in colorectal cancer tissues, para cancer tissues, SW480 cells, and CCC-HIE-2 cells.SW480 cells were divided into siRNA NC group, KCNQ1OT1 siRNA group, PAK4 siRNA group, inhibitor NC group, miR-145 inhibitor group, pcDNA-3.1(+)+mimics NC group, pcDNA-KCNQ1OT1+mimics NC group, pcDNA-3.1(+)+miR-145 mimics group, pcDNA-KCNQ1OT1+miR-145 mimics group, the expressions of KCNQ1OT1, miR-145 and PAK4 were knocked down, respectively. Cell viability was analyzed by MTT assay, the number of clones was analyzed by cell colony formation assay, cell apoptosis rate was detected by flow cytometry, and the EMT ability of SW480 cells was detected by Western blot. Result: Compared with para cancer tissues (1.37±0.27), LncRNA KCNQ1OT1 expression in colorectal cancer tissues was up-regulated (2.32±0.89) (t=23.84, P<0.01). The expression of PAK4 in colorectal cancer tissues was up-regulated (2.03±0.54) compared with paracellular tissues (1.21±0.09) (t=20.75, P<0.01). Compared with para cancer tissues (1.26±0.35), the expression of miR-145 in colorectal cancer tissues was significantly down-regulated (0.45±0.13) (t=18.76, P<0.01). Compared with CCC-HIE-2 cells (1.05±0.21), LncRNA KCNQ1OT1 expression in SW480 cells was up-regulated (2.09±0.27) (t=18.79, P<0.01). Compared with CCC-HIE-2 cells (1.18±0.62), PAK4 expression in SW480 cells was up-regulated (2.58±0.82) (t=22.17, P<0.01). Compared with CCC-HIE-2 cells (1.46±0.28), the expression of miR-145 in SW480 cells was significantly down-regulated (0.58±0.23) (t=17.86, P<0.01). LncRNA KCNQ1OT1 targets miR-145 and MmiR-145 targets PAK4. After KCNQ1OT1 or PAK4 expression was decreased, SW480 cell viability, clone number, and EMT ability were significantly decreased, and the apoptosis rate was up-regulated. Upregulation of KCNQ1OT1 targeting miR-145 promoted proliferation and EMT of SW480 cells and down-regulated apoptosis rate. Conclusion: Down-regulating LncRNA KCNQ1OT1 inhibited SW480 cell proliferation and EMT through the miR-145/PAK4 axis, and promoted cancer cell apoptosis

Key words： LncRNA KCNQ1OT1 miR-145 PAK4 Colorectal cancer Proliferation Apoptosis

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2023.02.02 OR http://www.hbyxzzs.cn/EN/Y2023/V29/I2/183

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