The Effects of FGD5-AS1 on the Proliferation and Apoptosis of Glioma Cells by Regulating the miR-3163/CNPY2 Axis
YANG Jiao1, HU Yuhua2*, XU Yueqiao3, LU Wenpeng2, ZHOU Chuan1, YU Zhuang1
This study aims to investigate the effects of the LncRNA (long non‑coding RNA) FGD5‑AS1 (FGD5 antisense RNA 1) on the proliferation and apoptosis of glioma cells by regulating the miR‑3163 (microRNA‑3163)/CNPY2 (canopy FGF signaling regulator 2) axis. The expression levels of LncRNA FGD5‑AS1, miR‑3163, and CNPY2 mRNA in normal human astrocytes and glioma cells (U87, LN229, U251) were detected by qRT‑PCR. U87 cells were divided into the following groups: control (NC) group, sh‑NC group, sh‑FGD5‑AS1 group, sh‑FGD5‑AS1+anti‑NC group, sh‑FGD5‑AS1+anti‑miR‑3163 group, miR‑NC group, miR‑3163 mimic group, miR‑3163 mimic+OE‑NC group, and miR‑3163 mimic+OE‑CNPY2 group. Cell proliferation and apoptosis were assessed by colony formation assay and flow cytometry, respectively. The protein expression levels of CNPY2, CCND1, and Bax were measured by Western blot. The targeting relationships among LncRNA FGD5‑AS1, miR‑3163, and CNPY2 were verified by RIP (RNA immunoprecipitation) and dual‑luciferase reporter assays. Compared with normal human astrocytes, the expression levels of LncRNA FGD5‑AS1 and CNPY2 mRNA were significantly increased in U87, LN229, and U251 cells, while miR‑3163 expression was significantly decreased (P<0.05). Among these, U87 cells showed the most pronounced changes in all three indicators. Compared with the sh‑NC group or miR‑NC group, the sh‑FGD5‑AS1 group or miR‑3163 mimic group exhibited significantly reduced colony formation numbers and CNPY2 and CCND1 protein expression levels, along with significantly increased apoptosis rates and Bax protein expression levels (P<0.05). Compared with the sh‑FGD5‑AS1+anti‑NC group or miR‑3163 mimic+OE‑NC group, the sh‑FGD5‑AS1+anti‑miR‑3163 group or miR‑3163 mimic+OE‑CNPY2 group showed significantly increased colony formation numbers and CNPY2 and CCND1 protein expression levels, as well as significantly decreased apoptosis rates and Bax protein expression levels (P<0.05). RIP and dual‑luciferase reporter assays confirmed the targeting relationships between miR‑3163 and LncRNA FGD5‑AS1, as well as between miR‑3163 and CNPY2 (P<0.05). Knockdown of LncRNA FGD5‑AS1 can inhibit glioma cell proliferation and promote apoptosis by upregulating miR‑3163 and downregulating CNPY2.
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