抑制circRNA ABCB10的表达对结直肠癌细胞生物学特性的影响

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目的:探讨circABCB10在结直肠癌组织和细胞中的表达及其对细胞生物学行为、放射敏感性和裸鼠皮下移植瘤生长的影响。方法:收集2018年1月至2018年12月于河南省人民医院治疗的结直肠癌患者的结直肠癌组织和癌旁正常组织。采用定量聚合酶链反应(qPCR)检测结直肠癌组织、癌旁正常组织和结直肠癌细胞SW480、HCT116、HT29中circABCB10和miR-217的表达;四甲基偶氮唑蓝(MTT)法检测细胞活力,流式细胞术检测细胞凋亡情况,Transwell法检测细胞迁移和侵袭能力,克隆形成实验检测放射敏感性,Circular RNA Interactome预测circABCB10下游miRNAs的表达,双荧光素酶报告基因实验进一步验证,裸鼠皮下移植瘤实验检测circABCB10对移植瘤生长的影响。结果:结直肠癌组织中circABCB10 mRNA的表达水平(3.97±2.12)高于癌旁组织(1.13±0.64,n P<0.05)。正常结肠上皮细胞FHC中circABCB10 mRNA的表达水平(1.00±0.09)低于结直肠癌细胞SW480、HCT116、HT29(分别为4.53±0.44、3.12±0.32和3.51±0.36,均n P<0.05)。MTT检测结果显示,转染48、72 h,si-circABCB10-1组SW480细胞的吸光度值分别为0.36±0.04和0.43±0.04,低于circ-NC组(分别为0.48±0.05和0.82±0.08,均n P<0.05)。si-circABCB10-1组SW480细胞的迁移细胞数[(45±8)个]和侵袭细胞数[(34±7)个]均低于circ-NC组[分别为(106±21)个和(84±15)个,均n P<0.01],放射增敏比为1.632。裸鼠皮下移植瘤实验结果显示,8 d后,si-circABCB10-1组肿瘤体积和重量低于circ-NC组(n P<0.05)。miR-217是circABCB10的靶基因,抑制miR-217的表达可逆转抑制circABCB10对细胞增殖、迁移、侵袭和裸鼠皮下移植瘤生长的抑制作用及放射增敏作用。n 结论:抑制circABCB10的表达可通过上调miR-217的表达来抑制结直肠癌SW480细胞的增殖、迁移和侵袭能力以及裸鼠皮下移植瘤的生长,并增加细胞的放射敏感性,揭示了结直肠癌进展的潜在分子机制,可为临床结直肠癌放射治疗提供一个新的增敏靶点。“,”Objective:To investigate the expression of circular ribonucleic acid ABCB10 (circABCB10) in colorectal cancer tissues and cells and its effects on cell biological behavior, radiosensitivity and growth of subcutaneous xenografts.Methods:The tumor tissue and adjacent tissue from colorectal cancer patients treated in Henan People′s Hospital were collected from January 2018 to December 2018. Quantitative polymerase chain reaction (qPCR) was used to detect the expressions of circABCB10 and miR-217, cell viability was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT), cell apoptosis rate was detected by flow cytometry, cell migration and invasion were detected by Transwell method, cell radiosensitivity was detected by colony formation assay. The downstream miRNAs of circABCB10 were predicted by Circular RNA Interactome and verified by the dual luciferase reporter gene experiment. The effect of circABCB10 on the growth of transplanted tumor was examined in nude mice.Results:The expression level of circABCB10 mRNA in colorectal cancer tissues was (3.97±2.12), higher than (1.13±0.64) in adjacent tissues (n P<0.05). The expression level of circABCB10 mRNA in FHC cells was (1.00±0.09), lower than that (4.53±0.44) in SW480, (3.12±0.32) in HCT116 and (3.51±0.36) in HT29 cells, respectively (alln P<0.05). The MTT results showed that the absorbance values of SW480 cells in si-circABCB10-1 group at 48 and 72 hours after transfection were (0.36±0.04) and (0.43±0.04), lower than (0.48±0.05) and (0.82±0.08) in circ-negative control (NC) group, respectively (alln P<0.05). The number of migrating cells and invasive cells in si-circABCB10-1 group were (45±8) and (34±7), lower than (106±21) and (84±15) in circ-NC group, respectively (alln P<0.01). The radiosensitization ratio was 1.632. The results of subcutaneous transplantation assay showed that the tumor volume and tumor weight of the si-circABCB10-1 group were significantly lower than circ-NC group after 8 days of inoculation ( alln P<0.05). MiR-217 is a target gene of circABCB10. Inhibition of miR-217 reversed the inhibitory effect of circABCB10 silencing on cell proliferation, migration, invasion and subcutaneous xenograft growth in nude mice and the radiosensitization activity.n Conclusion:Silence of circABCB10 can up-regulate the expression of miR-217 to inhibit the proliferation, migration, invasion and growth of subcutaneous xenografts and increase the radiosensitivity of SW480 cells, which reveals the underlying molecular mechanism of colorectal cancer progression and provides a new sensitizing target for clinical radiotherapy of colorectal cancer.
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