Journal: International Journal of Molecular Medicine

Article Title: CircRAD18 promotes glioblastoma proliferation, migration and invasion via the miR-1231/LUC7L2 axis

doi: 10.3892/ijmm.2026.5851

Figure Lengend Snippet: CircRAD18 acts as a sponge for miR-1231 in GBM cells. (A) Venn diagram showing miRNA targets of circRAD18 predicted using the CircBank and CircInteractome databases. (B) Volcano plot showing differentially expressed miRNAs in GBM cells, including 373 upregulated and 807 downregulated. (C) Venn diagram illustrating the overlap between predicted miRNAs and downregulated miRNAs from the GSE90603 dataset. (D) Data from the CGGA database showing that miR-1231 expression decreased as glioma grade increased. (E) Using data from the CGGA database, patients were categorized by miR-1231 levels and generated Kaplan-Meier survival curves to compare survival rates between the two groups. (F) FISH was used to detect the co-localization of circRAD18 and miR-1231 in the cytoplasm of GBM cells. circRAD18 is shown in red, miR-1231 in green, and nuclei are stained blue with DAPI. Scale bar, 20 μ m. Nucleocytoplasmic fractionation analysis followed by RT-qPCR demonstrated that miR-1231 was predominantly localized in the cytoplasm of (G) U87 and (H) U251 cells. β-actin and U6 were used as cytoplasmic and nuclear markers, respectively, to validate the fractionation efficiency. (I) Predicted binding sites of miR-1231 in circRAD18, with the Mut version of circRAD18 presented. (J) The dual-luciferase reporter assay determining the binding between circRAD18 and miR-1231. (K) RNA pull-down assay performed to verify the direct binding between circRAD18 and miR-1231. (L) RT-qPCR showed that circRAD18 knockdown markedly increased miR-1231 expression in U87 and U251 cells. All data are presented as mean ± SEM and each experiment was performed in triplicate. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. circ, circular RNA; GBM, glioblastoma; miRNA, microRNA; CGGA, Chinese Glioma Genome Atlas; FISH, fluorescence in situ hybridization; RT-qPCR, reverse transcription-quantitative PCR; MUT, mutant; WT, wild type.

Article Snippet: The small interfering RNAs (siRNAs) investigated in the present study included si1-circRAD18, si2-circRAD18, circRAD18-negative control (NC), miR-1231 mimic, miR-1231 mimic NC, miR-1231 inhibitor and miR-1231 inhibitor NC, all designed and synthesized by General Biosystems (Anhui) Co., Ltd.

Techniques: Expressing, Generated, Staining, Fractionation, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Pull Down Assay, Knockdown, Fluorescence, In Situ Hybridization, Reverse Transcription, Real-time Polymerase Chain Reaction, Mutagenesis

Journal: International Journal of Molecular Medicine

Article Title: CircRAD18 promotes glioblastoma proliferation, migration and invasion via the miR-1231/LUC7L2 axis

doi: 10.3892/ijmm.2026.5851

Figure Lengend Snippet: miR-1231 directly targets LUC7L2. (A) Venn diagram showing predicted miR-1231 target mRNAs using TargetScan, miRWalk and mirDIP databases. (B) Volcano plot showing differentially expressed mRNAs in GBM cells, with 2,499 upregulated and 3,324 downregulated. (C) Venn diagram illustrating the overlap between predicted mRNAs and upregulated mRNAs from the GSE4290 dataset. (D) Data from the CGGA database indicating that LUC7L2 expression increased with glioma grade. (E) Using data from the CGGA database, patients were categorized by LUC7L2 levels and generated Kaplan-Meier survival curves to compare the survival rates between the two groups. (F) Predicted binding sites of miR-1231 in LUC7L2 3'-UTR, with the Mut version of LUC7L2 3'-UTR presented. (G) The dual-luciferase reporter assay confirming the binding between miR-1231 and LUC7L2. (H) Western blot analysis showing that miR-1231 mimic treatment reduced LUC7L2 protein levels in U87 and U251 cells. All data are presented as mean ± SEM and each experiment was performed in triplicate. * P<0.05; ** P<0.01; *** P<0.001. miRNA, microRNA; GBM, glioblastoma; CGGA, Chinese Glioma Genome Atlas; NC, negative control; MUT, mutant; WT, wild type.

Article Snippet: The small interfering RNAs (siRNAs) investigated in the present study included si1-circRAD18, si2-circRAD18, circRAD18-negative control (NC), miR-1231 mimic, miR-1231 mimic NC, miR-1231 inhibitor and miR-1231 inhibitor NC, all designed and synthesized by General Biosystems (Anhui) Co., Ltd.

Techniques: Expressing, Generated, Binding Assay, Luciferase, Reporter Assay, Western Blot, Negative Control, Mutagenesis

Journal: International Journal of Molecular Medicine

Article Title: CircRAD18 promotes glioblastoma proliferation, migration and invasion via the miR-1231/LUC7L2 axis

doi: 10.3892/ijmm.2026.5851

Figure Lengend Snippet: circRAD18 promotes the proliferation, migration and invasion of GBM cells by sponging miR-1231. CCK-8 assays showing that miR-1231 inhibitor transfection promoted the proliferation of (A) U87 and (B) U251 cells. Wound healing assays showing that miR-1231 inhibitor transfection enhanced the migration of (C) U87 and (D) U251 cells. Scale bar, 100 μ m. (E) Transwell assays showed that miR-1231 inhibitor transfection promoted the invasion of U87 and U251 cells. Scale bar, 100 μ m. CCK-8 assays showing that si-circRAD18 co-transfection attenuated the proliferation-promoting effect of miR-1231 inhibitor in (F) U87 and (G) U251 cells. Wound healing assays showing that si-circRAD18 co-transfection attenuated the migration-promoting effect of miR-1231 inhibitor in (H) U87 and (I) U251 cells. Scale bar, 100 μ m. (J) Transwell assays showing that si-circRAD18 co-transfection attenuated the invasion-promoting effect of miR-1231 inhibitor in U87 and U251 cells. Scale bar, 100 μ m. All data are presented as mean ± SEM and each experiment was performed in triplicate. * P<0.05; *** P<0.001; **** P<0.0001. circ, circular RNA; GBM, glioblastoma; miRNA, microRNA; si, small interfering.

Article Snippet: The small interfering RNAs (siRNAs) investigated in the present study included si1-circRAD18, si2-circRAD18, circRAD18-negative control (NC), miR-1231 mimic, miR-1231 mimic NC, miR-1231 inhibitor and miR-1231 inhibitor NC, all designed and synthesized by General Biosystems (Anhui) Co., Ltd.

Techniques: Migration, CCK-8 Assay, Transfection, Cotransfection

Journal: International Journal of Molecular Medicine

Article Title: CircRAD18 promotes glioblastoma proliferation, migration and invasion via the miR-1231/LUC7L2 axis

doi: 10.3892/ijmm.2026.5851

Figure Lengend Snippet: circRAD18 promotes the proliferation, migration and invasion of GBM cells through the miR-1231/LUC7L2 axis. (A) Western blot analysis showing that circRAD18 knockdown reduced LUC7L2 protein levels in U87 and U251 cells. (B) Western blot analysis showing that miR-1231 inhibitor increased LUC7L2 protein levels in U87 and U251 cells. (C) Western blot analysis showing that si-circRAD18 co-transfection attenuated the increase in LUC7L2 protein levels induced by miR-1231 inhibitor in U87 and U251 cells. (D) Western blot analysis showing that LUC7L2 overexpression increased LUC7L2 protein levels in U87 and U251 cells. (E) RT-qPCR analysis showing that LUC7L2 overexpression increased LUC7L2 expression in U87 and U251 cells. CCK-8 assays showing that LUC7L2 overexpression promoted the proliferation of (F) U87 and (G) U251 cells. Wound healing assays showing that LUC7L2 overexpression enhanced the migration of (H) U87 and (I) U251 cells. Scale bar, 100 μ m. (J) Transwell assays showing that LUC7L2 overexpression promoted the invasion of U87 and U251 cells. Scale bar, 100 μ m. CCK-8 assays showing that circRAD18 knockdown attenuated the proliferation-promoting effect of LUC7L2 overexpression in (K) U87 and (L) U251 cells. Wound healing assays showing that circRAD18 knockdown attenuated the migration-promoting effect of LUC7L2 overexpression in (M) U87 and (N) U251 cells. Scale bar, 100 μ m. (O) Transwell assays showed that circRAD18 knockdown attenuated the invasion-promoting effect of LUC7L2 overexpression in U87 and U251 cells. Scale bar, 100 μ m. All data are presented as mean ± SEM and each experiment was performed in triplicate. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. circ, circular RNA; GBM, glioblastoma; miRNA, microRNA; si, small interfering; RT-qPCR, reverse transcription-quantitative PCR; OE, overexpressed; NC, negative control.

Article Snippet: The small interfering RNAs (siRNAs) investigated in the present study included si1-circRAD18, si2-circRAD18, circRAD18-negative control (NC), miR-1231 mimic, miR-1231 mimic NC, miR-1231 inhibitor and miR-1231 inhibitor NC, all designed and synthesized by General Biosystems (Anhui) Co., Ltd.

Techniques: Migration, Western Blot, Knockdown, Cotransfection, Over Expression, Quantitative RT-PCR, Expressing, CCK-8 Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: CircRAD18 promotes glioblastoma proliferation, migration and invasion via the miR-1231/LUC7L2 axis

doi: 10.3892/ijmm.2026.5851

Figure Lengend Snippet: circRAD18 knockdown inhibits GBM progression in vivo . (A and B) Representative images of xenograft tumors in nude mice showing that circRAD18 knockdown reduced tumor growth. (C) Tumor growth curves showing that circRAD18 knockdown slowed the growth of subcutaneous xenograft tumors. (D) Tumor volume measurements at day 28 showing that circRAD18 knockdown reduced tumor size compared with the control group. (E) Tumor weights measured on day 28 showing that circRAD18 knockdown decreased tumor mass compared with the control group. (F) RT-qPCR analysis showing that circRAD18 knockdown decreased circRAD18 expression in xenograft tumors. (G) RT-qPCR analysis showing that circRAD18 knockdown increased miR-1231 expression in xenograft tumors. (H) RT-qPCR analysis showing that circRAD18 knockdown decreased LUC7L2 expression in xenograft tumors. (I) IHC staining of xenograft tumors showing that circRAD18 knockdown decreased LUC7L2 protein levels. Scale bar, 100 μ m. All data are presented as mean ± SEM and each experiment was performed in triplicate. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001. circ, circular RNA; GBM, glioblastoma; IHC, immunohistochemistry; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin.

Article Snippet: The small interfering RNAs (siRNAs) investigated in the present study included si1-circRAD18, si2-circRAD18, circRAD18-negative control (NC), miR-1231 mimic, miR-1231 mimic NC, miR-1231 inhibitor and miR-1231 inhibitor NC, all designed and synthesized by General Biosystems (Anhui) Co., Ltd.

Techniques: Knockdown, In Vivo, Control, Quantitative RT-PCR, Expressing, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Proliferation, survival, and invasion phenotypes were reshaped by gain- and loss-of-function perturbation. (A) WB analysis of CENPI protein in HCC cell lines (HCCLM3, Hep3B, and HepG2); HepG2 showed the highest CENPI expression ( n = 3). (B) WB analysis of CENPI protein in si-CENPI-transfected HepG2 cells; si-CENPI decreased CENPI levels vs si-NC, *** p < 0.001 ( n = 3). (C) WB analysis of CENPI protein in oe-CENPI-transfected HepG2 cells; oe-CENPI increased CENPI levels vs oe-NC, *** p < 0.001 ( n = 3). (D) CCK-8 assay for HepG2 proliferation; si-CENPI decreased absorbance (weaker proliferation) vs si-NC, while oe-CENPI increased it (stronger proliferation) vs oe-NC, * p < 0.05 ( n = 3). (E) Wound-healing assay for HepG2 migration; healing status recorded at 0/24 h. si-CENPI decelerated wound closure vs si-NC, while oe-CENPI accelerated it vs oe-NC; scale bar = 100 μm, ** p < 0.01 ( n = 3). (F) Flow cytometry for HepG2 apoptosis; apoptotic rates quantified. si-CENPI increased apoptotic rate vs si-NC, while oe-CENPI decreased it vs oe-NC, ** p < 0.01 ( n = 3). (G) Transwell assay for HepG2 invasion in Matrigel-coated chambers; invaded cells stained and counted. si-CENPI reduced invaded cells vs si-NC, while oe-CENPI increased them vs oe-NC; scale bar = 275 μm, *** p < 0.001 ( n = 3). (H) Flow cytometry for HepG2 cell cycle; si-CENPI increased the proportion of cells in G1 phase and decreased that in S phase vs si-NC, ** p < 0.01, while oe-CENPI showed the opposite effect vs oe-NC, * p < 0.05 ( n = 3). Data are presented as mean ± SD of three independent experiments; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Expressing, Transfection, CCK-8 Assay, Wound Healing Assay, Migration, Flow Cytometry, Transwell Assay, Staining

Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Cell cycle and mTORC1 programs emerged as dominant signatures accompanied by CDK2 elevation and EMT marker remodeling. (A) KEGG pathway enrichment analysis (top ten significantly enriched pathways); the cell cycle pathway was most significant, with significance represented by −log10 ( p value), based on CENPI-related differentially expressed genes. (B) GSEA analysis of CENPI-related genes; enriched pathways include cell cycle-related (E2F_TARGETS) and mechanism-related (mTORC1_SIGNALING), significance represented by normalized enrichment score (NES). (C) WB analysis of cell cycle-related proteins (CDK2, Cyclin D1) and CENPI in si-CENPI and oe-CENPI-transfected HCC cells; CDK2 showed the most obvious change, ** p < 0.01 vs si-NC and * p < 0.05 vs oe-NC ( n = 3). (D) WB detection of PI3K/AKT/mTOR pathway proteins (PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR), EMT proteins (N-cadherin, E-cadherin, Vimentin), and CENPI in si-CENPI and oe-CENPI-transfected HCC cells, * p < 0.05 and ** p < 0.01 ( n = 3). Quantitative data are presented as mean ± SD of three independent experiments; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Marker, Transfection

Journal: Cancer Biology & Therapy

Article Title: Centromere protein I promotes hepatocellular carcinoma progression by activating PI3K/AKT/mTOR-CDK2 cascade

doi: 10.1080/15384047.2026.2667596

Figure Lengend Snippet: Orthotopic growth suppression was accompanied by pathway deactivation and reversal of EMT marker directionality. (A and B) Quantification of tumor weight in orthotopic xenograft models; shRNA-mediated CENPI silencing reduced tumor weight compared with models; scale bar = 1 cm, (C) Assessment of body weight changes; CENPI silencing attenuated cachexia-driven body weight loss, *** p < 0.001 vs control and ### p < 0.001 vs model ( n = 6). (D) Measurement of final tumor mass; CENPI silencing decreased tumor mass vs models without overt hepatotoxicity, ** p < 0.01 vs model ( n = 6). (E) WB analysis of EMT markers (E-cadherin, N-cadherin, and Vimentin) and PI3K/AKT/mTOR-CDK2 pathway proteins in resected tumor tissues. CENPI silencing induced a mesenchymal-to-epithelial reverting signature, with E-cadherin increased and N-cadherin/Vimentin suppressed. Concomitantly, p-PI3K, p-AKT, p-mTOR, and total CDK2 levels were diminished, indicating inactivation of the PI3K/AKT/mTOR-CDK2 relay, * p < 0.05 vs control, # p < 0.05, and ## p < 0.01 vs model ( n = 3). Data are presented as mean ± SD; error bars represent SD.

Article Snippet: CENPI-targeting siRNAs and a nontargeting control siRNA (si-NC) were synthesized by Sangon Biotech (Shanghai, China).

Techniques: Marker, shRNA, Control