超级增强子长链非编码RNA LINC01232在大肠癌组织中的表达及其对癌细胞生物学行为的影响
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国家自然科学基金(81871718);江苏省医学创新团队(CXTDA2017017)。


Expression of super-enhancer-associated long non-coding RNA LINC01232 in colorectal cancer tissues and its effects on cancer cell biological behavior
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    摘要:

    目的?探讨超级增强子长链非编码RNA(SE-LncRNA)LINC01232在大肠癌组织中的表达水平及其对大肠癌细胞生物学行为的影响。方法?通过基因芯片筛选4对大肠癌患者组织(癌组织和癌旁组织)中差异表达的SE-LncRNA,发现LINC01232在癌组织中显著高表达(Fold Change=2.656 9,P=0.025 8);采用实时荧光定量PCR(RT-qPCR)检测LINC01232在31对大肠癌组织中的表达水平,并分析其与患者临床病理参数的关系;应用ROC曲线分析LINC01232对大肠癌诊断的敏感性和特异性;将2种人大肠癌细胞系HCT-116和HT-29分别设置对照组(NC)和转染LINC01232 Smarter Silence组(si-LINC01232),通过CCK-8试验、细胞克隆形成试验及Transwell试验检测LINC01232对大肠癌细胞增殖、侵袭和迁移等生物学行为的影响;对跨膜蛋白超家族9-2(TM9SF2)与LINC01232在组织中的表达量进行相关性分析,并采用western blot检测下调LINC0123对TM9SF2蛋白表达的影响。结果?LINC01232在大肠癌组织中的表达水平显著高于癌旁组织,差异具有统计学意义[(2.015±2.865) vs (1.000±2.036),t=2.388,P=0.023];LINC01232在组织中的表达量与TNM临床分期(χ2=5.427,P=0.020)和远处转移(χ2=4.663,P=0.031)有关;ROC曲线分析结果显示,LINC01232对大肠癌诊断的敏感性为53.13%,特异性为78.13%;与NC组相比,si-LINC01232组细胞的增殖、克隆形成率、侵袭及迁移能力显著降低(P<0.05);在大肠癌组织中,TM9SF2与LINC01232的表达呈正相关(r=0.51,P<0.01),而在大肠癌细胞中,TM9SF2?mRNA和蛋白质水平在LINC01232下调后降低(P<0.05)。结论?LINC01232在大肠癌组织中异常高表达,并促进大肠癌细胞的增殖、侵袭和迁移,有望成为大肠癌治疗的新靶点。

    Abstract:

    Objective?To investigate the expression levels of super-enhancer-associated long non-coding RNA (SE-LncRNA) LINC01232 in colorectal cancer (CRC) tissues and its effects on biological behavior of CRC cells.?Methods?Four paired differentially expressed SE-LncRNAs from colorectal cancer tissues (cancer and paracancerous tissues) were screened by gene microarray, and LINC01232 was.and LINC01232 (Fold Change=2.656 9,?P=0.025 8) was identified to be overexpressed in cancer tissues for further investigation. RT-qPCR was performed to detect the expression of LINC01232 in 31 paired CRC tissues, and the correlation with clinic pathological characteristics was analyzed. The sensitivity and specificity of LINC01232 in CRC diagnosis were analyzed by ROC curve., The control group (NC) and smarter silence transfected group (si-LINC01232) were set in two human colorectal cancer cell lines HCT-116 and HT-29 cells, respectively. The effects of LINC01232 on CRC cell proliferation, migration and invasion were studied by CCK-8 assay, colony formation assay and transwell assay. The correlation between TM9SF2 and LINC01232 expression in CRC tissues was analyzed, and the expression of TM9SF2 protein was detected by western blot.?Results?LINC01232 was significantly overexpressed in CRC tissues when compared with adjacent none-tumor tissues [(2.015±2.865) vs (1.000±2.036),?t=2.388,?P=0.023]. LINC01232 was positively correlated with TNM stage (χ2=5.427,P=0.020) and distant metastasis (χ2=4.663,?P=0.031). ROC curve showed the sensitivity and specificity of LINC01232 in CRC diagnosis were 53.13% and 78.13%, respectively. Compared with NC group, the cell proliferation, clone formation rate, migration and invasion ability of si-LINC01232 group were significantly reduced (P<0.05). In CRC tissues, the expression of TM9SF2 was positively correlated with LINC01232 (r=0.51,?P<0.01). In CRC cell lines, the RNA and protein levels of TM9SF2 were significantly decreased (P<0.05), when LINC01232 was down-regulated.?Conclusion?The expression of LINC01232 was significantly upregulated in CRC tissues, and promoted CRC cell proliferation, migration and invasion, suggesting LINC01232 may be become a new target for CRC treatment.

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陈缓缓,郑钧予,江盼,严枫.超级增强子长链非编码RNA LINC01232在大肠癌组织中的表达及其对癌细胞生物学行为的影响[J].临床检验杂志,2021,39(3):165-171

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  • 收稿日期:2020-12-08
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  • 在线发布日期: 2021-05-17
  • 出版日期: 2021-03-28
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