Inhibitory effects of guanylate binding proteins 5 on proliferation,invasion and migration of color-ectal cancer cells
Objective To investigate the biological function and potential molecular mechanism of guanylate binding protein 5(GBP5)on colorectal cancer(CRC)cells.Methods CRC and paraneoplastic tissues from the Xijing Hospital of Fourth Military Medical University were selected for the study,and the expression of GBP5 in these tissues were detected by real time quantitative polymerase chain reaction(RT-qPCR)and immunohistochemistry.5-Ethynyl-2'-deoxyuridine(EdU),colony formation assays,the flow cytometry,and Transwell assay were used to verify the biological function of GBP5 in CRC cells.Transcriptome sequencing combined with bioinformatics analysis was used to explore the mechanism of GBP5 inhibiting the malignant progression of CRC cells.The t-test was used for the comparison of the measurement data between the two groups,and ANOVA was used to compare between multiple groups.Results The expression level of GBP5 in part of CRC tissues was significantly lower than that of paired normal tissues(1#:0.09±0.01 vs.1.00±0.40,t=3.992,P＜0.05;2#:0.29±0.20 vs.1.00± 0.23,t=3.882,P＜0.05;3#:0.12±0.02vs.1.00±0.44,t=3.493,P＜0.05;15#:0.11±0.04 vs.1.00±0.18,t=8.265,P＜0.05).Patients in the GBP5 high expression group had higher survival rates(Log rank P＜0.05).The proportion of EdU-positive and S-phase cells in GBP5 knockdown group was significantly more than that in LV-shNC group[(49.9±7.5)％vs.(82.8±9.6)％,t=4.673,P＜0.05;(38.9±2.7)％vs.(49.1±1.8)％,t=5.441,P＜0.05].The number of cell colonies,cells crossing the matrigel and wells were more in GBP5 knockdown group than in LV-shNC group[(437.33± 24.58)vs.(737.00±124.94),t=4.076,P＜0.05;(103.00±7.94)vs.(172.00±6.92),t=11.34,P＜0.05;(329.33±103.25)vs.(898.33±133.38),t=5.843,P＜0.05].The apoptosis rate was lower in GBP5 knockdown group than that in LV-shNC group[(4.98±0.07)％vs.(2.11± 0.11)％,t=39.23,P＜0.05].The proportion of EdU-positive and S phase cells in LV-GBP5 group was significantly lower than that in LV-NC group[(67.4±9.8)％vs.(27.6±12.3)％,t=4.377,P＜0.05;(54.9±1.7)％vs.(49.3±2.7)％,t=3.100,P＜0.05].The number of cell colonies,cells crossing the matrigel and wells were less in LV-GBP5 group than in LV-shNC group[(1 299.67±173.28)vs.(991.00±6.56),t=3.083,P＜0.05;(113.67±19.86)vs.(57.33±16.62)％,t=3.768,P＜0.05;(279.66±15.57)vs.(132.67±63.31),t=3.905,P＜0.05].The apoptosis rate was higher in LV-GBP5 group than that in LV-NC group[(3.70±0.15)％vs.(4.65±0.25)％,t=5.689,P＜0.05].Conclusion GBP5 was downregulated in RCC.GBP5 overexpression could inhibit the prolifera-tion,invasion and migration ability and promote apoptosis in CRC cells.
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