Fig 1: DYNC1I1 upregulated TNPO2 expression by upregulating SP1 in gastric cancer cells. A, Transcription factors (TF) prediction in the promoter region of TNPO2 on GeneCards (only the top four) and ALGGEN PROMO website, respectively, intersect the results obtained by the two websites. B, Correlation between DYNC1I1 and SP1, TNPO2, and SP1 in gastric cancer were performed using GEPIA online platform; differences with P < .05 were considered statistically significant. C, SP1 mRNA expression levels were tested by qRT-PCR in HGC-27, SGC-7901, and SNU-216 cells after transfected with siDYNC1I1 for 48 h. D, SP1 protein expression was detected by Western blot analysis in HGC-27, SGC-7901, and SNU-216 cells after transfected with siDYNC1I1 for 48 h. E, SP1 and TNPO2 mRNA expression levels were tested by qRT-PCR in HGC-27, SGC-7901, and SNU-216 cells after transfected with siSP1 for 48 h. F, SP1 and TNPO2 protein expressions were detected by Western blot analysis in HGC-27, SGC-7901, and SNU-216 cells after transfected with siSP1 for 48 h. G, The luciferase activities in HGC-27 cotransfected with SP1 overexpression or NC and luciferase reporters containing TNPO2 promotor WT or TNPO2 promotor MUT. Student's t tests were used for statistical analyses (**P < 0.01, ***P < 0.001, ****P < 0.0001; n = 3, student s t test, means ± 95% CI)
Fig 2: Silencing TNPO2 in gastric cancer cells inhibited proliferation of gastric cancer cells and promoted apoptosis. A, TNPO2 protein expression was detected by Western blot analysis in HGC-27, SGC-7901, and SNU-216 cells after transfected with siTNPO2 for 48 h. B, Analysis of cell proliferation following TNPO2 knockdown in HGC-27, SGC-7901, and SNU-216 cells by MTT assay. C, The apoptotic rates of HGC-27 and SGC-7901 cells transfected with siNC or siTNPO2 for 48 h were visualized by flow cytometry. D, EdU incorporation assay. Seventy-two hours after gastric cancer cells were transfected with siTNPO2 or siNC, followed by incubation with EdU and Hoechst in sequence. Hoechst 33342 (blue) and EdU (green) represent cell nuclei and nuclei of proliferative cells, respectively. The percentages of the EdU-positive cells are presented (right). E, Apoptotic proteins were detected by Western blot analysis in HGC-27 cells after transfected with siTNPO2 for 48 h (*P < 0.05, **P < 0.01, ***P < 0.001; n = 3, student s t test, means ± 95% CI)
Fig 3: DYNC1I1 upregulated TNPO2 in gastric cancer cell. HGC-27, SGC-7901, and SNU-216 cell lines were transfected with siDYNC1I1 for 48 h. A, After knockdown of DYNC1I1 in HGC-27 cells, the top 10 upregulated genes reported by microarray genome-wide expression analysis. B, Correlation between DYNC1I1 and TNPO2 in gastric cancer was performed using GEPIA online platform; differences with P < 0.05 were considered statistically significant. C, YNC1I1 and TNPO2 mRNA expression levels were tested by qRT-PCR in HGC-27, SGC-7901, and SNU-216 cells. D, DYNC1I1 and TNPO2 protein expressions were detected by Western blot analysis in HGC-27, SGC-7901, and SNU-216 cells (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n = 3, student s t test, means ± 95% CI)
Fig 4: TNPO2 is overexpressed in GC tissues and patients with high TNPO2 expression have a poor prognosis. A, Analysis of TNPO2 expression using GEPIA Website. B, Kaplan-Meier Plotter analysis of the effect of TNPO2 on GC patients' survival
Fig 5: Overexpression of TNPO2 promoted gastric cancer cell proliferation and inhibited apoptosis. A, TNPO2 protein expression was detected by Western blot analysis in MGC-803 cells after transfected with overexpressing TNPO2 for 48 h. B, MTT shows cell viability of gastric cancer cells after overexpressing TNPO2 for 48, 72, and 96 h. C, The percentages of the EdU-positive cells after overexpressing TNPO2 for 72 h. D, The apoptotic rates of MGC-803 cells after overexpressing TNPO2 for 48 h (*P < 0.05, **P < 0.01, ****P < 0.0001; n = 3, student s t test, means ± 95% CI)
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