Fig 1: Circ_101341 was highly expressed in ccRCC tissues and cells. (A) The differential expression of several circRNAs in ccRCC tissues identified from the GSE100186 dataset was shown in the heat map. (B) Schematic diagram illustrated that circ_101341 was generated from EGLN3 pre-mRNA. (C) Circ_101341 was highly expressed in ccRCC from the GSE100186 dataset. (D) The expression of circ_101341 in ccRCC tissues (n=60) and normal tissues (n=60) was measured by qRT-PCR. (E) The overall survival curve was generated by Kaplan–Meier plot and analyzed by Log rank test. (F) The expression of circ_101341 in ccRCC cell lines (SW839, A498, Caki-1 and 786-O) was detected by qRT-PCR. (G) The circularity and stability of circ_101341 were detected using RNase R by qRT-PCR. *P < 0.05.
Fig 2: Construction of a ceRNA of lncRNA-miRNA-mRNA.(A) LncRNA GSEC bound to eight miRNAs. (B) The expression of lncRNA GSEC. (C) Kaplan–Meier survival of lncRNA GSEC. (D)Venn diagram identified the downstream targets in the TargetScan database and IHRLs. (E)The expression of EGLN3 in the GEPIA database. (F) The ceRNA of lncRNA GSEC/miR-873-3p/EGLN3 regulatory axis. ∗p < 0.05; ∗∗∗p < 0.001.
Fig 3: miR-4646-5p/ABHD16A/Lyso-PS stimulated activation of RhoA trigger LIMK/cofilin signaling to promote GC metastasis in vivo and clinic.A, B Lyso-PS concentration of the mice lung and liver metastasis tissues in subgroups were determined by LC/MS-MS (**p < 0.01). C The change of activated RhoA and RhoA, p-LIMK and LIMK, p-cofilin and cofilin in mice lung and liver metastasis tissues by Western blot. D The relative GPR34 and RhoA levels in gastric tumor tissues from the patients with metastases (M+, n = 20) were compared with the age- and sex-matched patients without metastases (M-, n = 20). E Immunohistochemical staining to detect ABHD16A, RhoA, HIF1A, GPR34, p-LIMK, and p-cofilin levels in metastatic GC tumors or in non- metastatic tumor (magnification, ×200; Scale bar, 100 µm). F Summary chart of ABHD16A regulating and activating RhoA and its downstream signaling. RhoA-mediated GC metastasis closely relates with miR-4646-5p/PHD3/HIF1A mediated upregulation of RhoA and lyso-PS/GPR34 depended activation of RhoA in GC cells.
Fig 4: The expression and the effect of lncRNA GSEC, miR-873-3p, and EGLN3 in LUAD.(A) Expression of GSEC in different LUAD cell lines compared to normal astrocytes. (B) Expression of miR-873-3p in different LUAD cell lines compared to normal astrocytes. (C) Expression of EGLN3 in different LUAD cell lines compared to normal astrocytes. (D) Expression of EGLN3 at the western blot level. (E) The efficiency of GSEC knockdown (si-GSEC). (F-G) The proliferation of cells by CCK-8 assays. (H-I) The migration of cells by wound healing assays. (J-K) The cloning capacity of cells. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.
Fig 5: Schematic diagram of pro-metastatic function of miR-4646-5p/ABHD16A/lyso-PS axis in Drosha-low expressed gastric tumor.In Drosha-low expressed gastric cancer cells, mirtronic miR-4646-5p was abnormally up-regulated. The Elevated mirtronic miR-4646-5p is a specific splicing product of intron-3 of the host gene Abhd16a with the aid of enhanced SRSF2. The enhanced miR-4646-5p can mitigate ubiquitination of HIF1A by inhibiting the target gene PHD3, thus lead to an increased HIF1A. As a transcription factor, HIF1A feedback promotes the expressions of the host gene Abhd16a and miR-4646-5p itself. The Abhd16a encodes a lipase enzyme ABHD16A, which causes accumulation of lipid metabolite lyso-PS, and then activates RhoA through GPR34/Gi. As synergistic regulation, miRNA-4646-5p can also up-regulate RhoA expression through transcription factor HIF1A. The activation and up-regulation of RhoA synergistically promote gastric cancer metastasis through LIMK/cofilin signaling.
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