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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells <t>(HCT-15,</t> <t>p-RIP:</t> P=0.02; RIP: P=0.97; <t>p-MLKL:</t> P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.
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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells <t>(HCT-15,</t> <t>p-RIP:</t> P=0.02; RIP: P=0.97; <t>p-MLKL:</t> P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.
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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells <t>(HCT-15,</t> <t>p-RIP:</t> P=0.02; RIP: P=0.97; <t>p-MLKL:</t> P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.
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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells <t>(HCT-15,</t> <t>p-RIP:</t> P=0.02; RIP: P=0.97; <t>p-MLKL:</t> P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.
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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells <t>(HCT-15,</t> <t>p-RIP:</t> P=0.02; RIP: P=0.97; <t>p-MLKL:</t> P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.
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SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, p-RIP: P=0.02; RIP: P=0.97; p-MLKL: P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.

Journal: Translational Cancer Research

Article Title: SNORD93 suppresses colorectal cancer progression by inducing G0/G1 arrest and apoptosis

doi: 10.21037/tcr-2025-1236

Figure Lengend Snippet: SNORD93 inhibited the proliferation of CRC. (A,B) Comparison of the effect of SNORD93 -overexpression vector and control pGPU6-NC vector on the cell cycle of CRC cell lines via flow cytometry (HCT-15: P=0.001; HT-29: P=0.048). (C) Western blot profiling of cell cycle regulators in SNORD93 -overexpressing CRC cells (HCT-15, p21 Waf1/Cip1: P=0.004; cyclin E2: P=0.047; cyclin D3: P=0.04; CDK9: P=0.03) (HT-29, p21 Waf1/Cip1: P=0.007; cyclin E2: P=0.008; cyclin D3: P=0.03; CDK9: P=0.01). (D) Comparison of the effect of the SNORD93 -overexpression vector and the control pGPU6-NC vector on apoptosis in CRC cell lines via flow cytometry. (E) Western blot profiling of apoptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, caspase 8: P=0.044; caspase 3: P=0.044; caspase 7: P=0.006; Bim: P=0.01) (HT-29, caspase 8: P=0.01; caspase 3: P=0.02; caspase 7: P=0.008; Bim: P=0.002). (F) Western blot profiling of necroptosis markers in SNORD93 -overexpressing CRC cells (HCT-15, p-RIP: P=0.02; RIP: P=0.97; p-MLKL: P=0.04; MLKL: P=0.27) (HT-29, p-RIP: P=0.04; RIP: P=0.14; p-MLKL: P=0.01; MLKL: P=0.16). Data are presented as mean ± SD (n=3). ns, P>0.05; *, P<0.05; **, P<0.01. CRC, colorectal cancer; MLKL, mixed lineage kinase domain-like protein; NC, negative control; p-MLKL, phosphorylated MLKL; p-RIP, phosphorylated RIP; PI, propidium iodide; RIP, receptor-interacting protein; SD, standard deviation.

Article Snippet: Following this step, the membranes were blocked with 5% nonfat milk in Tris-buffered saline containing Tween-20 (TBST) for 2 hours at room temperature, which was followed by overnight incubation at 4 °C with the following primary antibodies: p21 [cat. No. 2947T; Cell Signaling Technology (CST), Danvers, MA, USA], cyclin D3 (cat. No. 2936T; CST), cyclin E2 (cat. No. 4132P; CST), CDK9 (cat. No. 2316P; CST), caspase 3 (cat. No. 14220T; CST), caspase 7 (cat. No. 12827T; CST), caspase 8 (cat. No. 4790T; CST), Bim (cat. No. 2933P; CST), receptor-interacting protein (RIP) (cat. No. 3493T; CST), phosphorylated RIP (p-RIP) (cat. No. 65746T; CST), mixed lineage kinase domain-like protein (MLKL) (cat. No. 14993T; CST), phosphorylated MLKL (p-MLKL) (cat. No. 91689T; CST), E-cadherin (cat. No. 3195T; CST), and β-catenin (cat. No. 8480T; CST). β-actin (cat. No. 3700; CST) and GAPDH (cat. No. 60004-1-Ig; Proteintech, Rosemont, IL, USA) were used as the reference proteins.

Techniques: Comparison, Over Expression, Plasmid Preparation, Control, Flow Cytometry, Western Blot, Negative Control, Standard Deviation