Journal: bioRxiv

Article Title: lncRNA-ISM1 Promotes Hepatocellular Carcinoma Progression through RBM10-Mediated Alternative Splicing of ISM1 and Akt-S6–Dependent Glucose Metabolic Reprogramming

doi: 10.64898/2026.02.27.708505

Figure Lengend Snippet: An ISM1-overexpressing lentivirus was transfected into MHCC-97H cells for 24 h. (A) Western blotting was used to assess the protein expression of ISM1, GLUT4, HK2, LDH, and PKM2, as well as the phosphorylation of Akt and S6. The statistical results are shown in (B) . Glucose consumption (C) , lactate production (D) , and ATP content (E) were measured in MHCC-97H cells. MHCC-97H cells were transfected with negative control (NC) siRNA and ISM1 siRNA (to silence ISM1) for 24 h. (F) qPCR was performed to assess the mRNA level of ISM1 in the cells. MHCC-97H cells were transfected with ISM1 siRNA for 24 h and then treated with MK2206 for another 24 h. (G) Western blotting was used to assess the protein expression of ISM1, GLUT4, HK2, LDH, and PKM2, as well as the phosphorylation of Akt and S6. The statistical results are shown in (H-I) . Glucose consumption (J) , lactate production (K) , and ATP content (L) were measured in the cells. The data are presented as the mean ± standard error of the mean (mean ± SEM). n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001; n.s., not significant.

Article Snippet: The following reagents were purchased or obtained from the following sources: rabbit anti-ISM1 antibody (PA5-24968, Invitrogen, USA), rabbit anti-ISM1-AS antibody (MR-YT-20221208-001, Mabioway, China), rabbit anti-GAPDH antibody (#2118s, CST, USA), rabbit anti-total AKT (#4685s, CST, USA), rabbit anti-phospho-AKT (Ser 473) (#4060t, CST, USA), rabbit anti-total S6 (#2217s, CST, USA), rabbit phospho-S6 (Ser 235/236) (#4858t, CST, USA), mouse anti-GLUT4 antibody (66846-1-Ig, Proteintech, China), rabbit anti-HK2 antibody (22029-1-AP, Proteintech, China), rabbit anti-LDH antibody (14546-1-AP, Proteintech, China), rabbit anti-PKM2 antibody (15822-1-AP, Proteintech, China), and rabbit anti-RBM10 (84104-2-RR, Proteintech, China).

Techniques: Transfection, Western Blot, Expressing, Phospho-proteomics, Negative Control

Journal: bioRxiv

Article Title: lncRNA-ISM1 Promotes Hepatocellular Carcinoma Progression through RBM10-Mediated Alternative Splicing of ISM1 and Akt-S6–Dependent Glucose Metabolic Reprogramming

doi: 10.64898/2026.02.27.708505

Figure Lengend Snippet: MHCC-97H cells were transfected with either a lncRNA-ISM1-overexpressing lentivirus or lncRNA-ISM1 siRNA. (A) qPCR was used to assess the mRNA level of lncRNA-ISM1 in the cells. (B) Western blotting was performed to assess the protein expression of ISM1-AS, ISM1, GLUT4, HK2, LDH, and PKM2, as well as the phosphorylation of Akt and S6. The statistical results are shown in (C) . Glucose consumption (D) , lactate production (E) , and ATP content (F) were measured in MHCC-97H cells. The data are presented as the mean ± standard error of the mean (mean ± SEM). n = 3. * p < 0.05, ** p < 0.01, and *** p < 0.001; n.s., not significant.

Article Snippet: The following reagents were purchased or obtained from the following sources: rabbit anti-ISM1 antibody (PA5-24968, Invitrogen, USA), rabbit anti-ISM1-AS antibody (MR-YT-20221208-001, Mabioway, China), rabbit anti-GAPDH antibody (#2118s, CST, USA), rabbit anti-total AKT (#4685s, CST, USA), rabbit anti-phospho-AKT (Ser 473) (#4060t, CST, USA), rabbit anti-total S6 (#2217s, CST, USA), rabbit phospho-S6 (Ser 235/236) (#4858t, CST, USA), mouse anti-GLUT4 antibody (66846-1-Ig, Proteintech, China), rabbit anti-HK2 antibody (22029-1-AP, Proteintech, China), rabbit anti-LDH antibody (14546-1-AP, Proteintech, China), rabbit anti-PKM2 antibody (15822-1-AP, Proteintech, China), and rabbit anti-RBM10 (84104-2-RR, Proteintech, China).

Techniques: Transfection, Western Blot, Expressing, Phospho-proteomics

Journal: bioRxiv

Article Title: lncRNA-ISM1 Promotes Hepatocellular Carcinoma Progression through RBM10-Mediated Alternative Splicing of ISM1 and Akt-S6–Dependent Glucose Metabolic Reprogramming

doi: 10.64898/2026.02.27.708505

Figure Lengend Snippet: (A) Datasets related to HCC, gene splicing, RBPs, glycometabolism, and antisense transcripts were downloaded from the GeneCards database ( https://www.genedcards.org ), and a Venn diagram was constructed to show the intersections. (B) qPCR was used to assess the mRNA levels of TUG1, BMP2, ATM, and RBM10 in cells overexpressing lncRNA-ISM1. (C) RNA immunoprecipitation assays were performed using an anti-RBM10 antibody in MHCC-97H cells. (D) After MHCC-97H cells were transfected with RBM10 siRNA for 24 h, qPCR was used to assess the mRNA level of RBM10 in the cells. (E) MHCC-97H cells overexpressing lncRNA-ISM1 were transfected with RBM10 siRNA, and Western blotting was used to assess the protein expression levels of ISM1-AS, ISM1, RBM10, GLUT4, HK2, LDH, and PKM2, as well as the phosphorylation levels of Akt and S6. The statistical results are shown in (F) . (G) EdU-based proliferation assays were performed using MHCC-97H cells treated as described in (E) . (H) Colony formation assays were performed using MHCC-97H cells treated as described in (E) for 14 days. (I) Transwell invasion assays were performed using MHCC-97H cells treated as described in (E) for 24 h. The data are presented as the mean ± standard error of the mean (mean ± SEM). n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Scale bar = 100 μm.

Article Snippet: The following reagents were purchased or obtained from the following sources: rabbit anti-ISM1 antibody (PA5-24968, Invitrogen, USA), rabbit anti-ISM1-AS antibody (MR-YT-20221208-001, Mabioway, China), rabbit anti-GAPDH antibody (#2118s, CST, USA), rabbit anti-total AKT (#4685s, CST, USA), rabbit anti-phospho-AKT (Ser 473) (#4060t, CST, USA), rabbit anti-total S6 (#2217s, CST, USA), rabbit phospho-S6 (Ser 235/236) (#4858t, CST, USA), mouse anti-GLUT4 antibody (66846-1-Ig, Proteintech, China), rabbit anti-HK2 antibody (22029-1-AP, Proteintech, China), rabbit anti-LDH antibody (14546-1-AP, Proteintech, China), rabbit anti-PKM2 antibody (15822-1-AP, Proteintech, China), and rabbit anti-RBM10 (84104-2-RR, Proteintech, China).

Techniques: Construct, RNA Immunoprecipitation, Transfection, Western Blot, Expressing, Phospho-proteomics

Journal: bioRxiv

Article Title: lncRNA-ISM1 Promotes Hepatocellular Carcinoma Progression through RBM10-Mediated Alternative Splicing of ISM1 and Akt-S6–Dependent Glucose Metabolic Reprogramming

doi: 10.64898/2026.02.27.708505

Figure Lengend Snippet: BALB/c nude mice (6 weeks old, weighing 18–22 g) were subcutaneously inoculated with human HCC MHCC-97H cells (2×10⁶ cells/mouse) on the right posterior back. The mice were divided into a control group, an NC group, an ISM1-overexpression group, an ISM1-knockdown group, a lncRNA-ISM1-overexpression group, and an ISM1-AS-overexpression group. The mice were observed for 4 weeks after inoculation, after which the tumours were dissected, photographed, and weighed. Representative photographs of tumour appearance and size on the backs of nude mice are shown in (A). (B) An analysis of tumour size is presented. Immunohistochemistry was used to assess the expression levels of P-AKT (C) , P-S6 (D) , and GLUT4 in mouse HCC tissues. (F) Western blotting was performed to assess the protein expression levels of ISM1, ISM1-AS, and GLUT4, as well as the phosphorylation levels of Akt and S6. The statistical results are shown in (G) . The data are presented as the mean ± standard error of the mean (mean ± SEM). n = 3-6. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Scale bar = 25 μm.

Article Snippet: The following reagents were purchased or obtained from the following sources: rabbit anti-ISM1 antibody (PA5-24968, Invitrogen, USA), rabbit anti-ISM1-AS antibody (MR-YT-20221208-001, Mabioway, China), rabbit anti-GAPDH antibody (#2118s, CST, USA), rabbit anti-total AKT (#4685s, CST, USA), rabbit anti-phospho-AKT (Ser 473) (#4060t, CST, USA), rabbit anti-total S6 (#2217s, CST, USA), rabbit phospho-S6 (Ser 235/236) (#4858t, CST, USA), mouse anti-GLUT4 antibody (66846-1-Ig, Proteintech, China), rabbit anti-HK2 antibody (22029-1-AP, Proteintech, China), rabbit anti-LDH antibody (14546-1-AP, Proteintech, China), rabbit anti-PKM2 antibody (15822-1-AP, Proteintech, China), and rabbit anti-RBM10 (84104-2-RR, Proteintech, China).

Techniques: Control, Over Expression, Knockdown, Immunohistochemistry, Expressing, Western Blot, Phospho-proteomics