| 林 冰,刘 沙,周 平.MiR-124对食管癌细胞增殖和侵袭的作用及机制研究[J].肿瘤学杂志,2020,26(12):1047-1051. |
| MiR-124对食管癌细胞增殖和侵袭的作用及机制研究 |
| The Effect of MiR-124 on Proliferation and Invasion of Esophageal Cancer Cells and Its Mechanism |
| 投稿时间：2019-12-03 |
| DOI：10.11735/j.issn.1671-170X.2020.12.B008 |
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| 中文关键词: 食管肿瘤 食管鳞状细胞癌 miR-124 CREB1 |
| 英文关键词:esophageal cancer esophageal squamous cell carcinoma miR-124 CREB1 |
| 基金项目: |
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| 中文摘要: |
| 摘 要：［目的］检测miR-124及其靶基因CREB1在食管癌组织中的表达情况，探讨miR-124/CREB1在人食管鳞癌细胞KYSE-150体外增殖和侵袭中的作用及机制。［方法］ 通过实时荧光定量PCR(qRT-PCR)检测32例食管鳞状细胞癌(ESCC)组织及周围正常组织的miR-124和CREB1表达。Targetscan分析miR-124与CREB1的结合位点，荧光素酶基因报告实验验证miR-124 与CREB1是否结合；过表达或抑制miR-124后，采用Western blot检测KYSE-150细胞中CREB1的表达；在KYSE-150细胞中转染miR-124 mimic为miR-124 mimic或转染miR-124抑制剂为anti-mi-R124，转染后24h、48h、72h收集细胞计数，检测细胞增殖情况；转染24h后通过体外侵袭实验检测过表达或抑制miR-124对KYSE-150细胞侵袭的影响。在KYSE-150细胞中转染CREB1 siRNA后24h、48h、72h收集细胞计数，检测细胞增殖情况；转染CREB1 siRNA 24h后检测KYSE-150细胞侵袭能力的变化；在KYSE-150细胞中共同转染anti-miR-124和siCREB1，检测细胞增殖和侵袭能力变化情况。［结果］ MiR-124在食管癌组织表达降低(P<0.001)，而CREB1在食管癌组织中表达升高(P<0.001)；CREB1是miR-124下游分子，过表达miR-124抑制KYSE-150细胞中CREB1的表达(P=0.016)，抑制miR-124促进KYSE-150细胞中CREB1的表达(P<0.001)。在食管癌细胞中，过表达miR-124抑制KYSE-150细胞的增殖和侵袭(P均<0.05)，抑制miR-124促进KYSE-150细胞的增殖和侵袭(P均<0.05)；敲低CREB1抑制KYSE-150细胞的增殖和侵袭(P均<0.05)。敲低CREB1可抑制anti-miR-124对KYSE-150细胞增殖和侵袭的促进作用(P均<0.05)。［结论］过表达miR-124通过靶向CREB1抑制食管癌细胞的增殖和侵袭。 |
| 英文摘要: |
| Abstract：［Objective］ To detect the expression of miR-124 and its target gene CREB1 in esophageal cancer tissues and evaluate the effect of miR-124 on proliferation and invasion of esophageal cancer cells and its mechanism. ［Methods］ The expression of miR-124 and CREB1 in esophageal cancer tissues and surrounding normal tissues in 32 cases of esophageal squamous cell(ESCC) was detected by real-time RT-PCR(qRT-PCR). The binding of miR-124 to CREB1 was analyzed by Targetscan and confirmed by luciferase gene reporter assay. The miR-124 mimic or anti-mi-R124 was transfected to human ESCC KYSE-150 cells，the expression of CREB1 in KYSE-150 cells was detected by Western blot after overexpression or inhibition of miR-124. The proliferation was detected at different time points(24，48，72h) after transfection，the effect of miR-124 on invasion ability of KYSE-150 cell was detected by in vitro invasion assay 24 h after transfection. Cell counts were performed in KYSE-150 cells at 24h，48h and 72h after transfection with CREB1 siRNA；the invasive ability of KYSE-150 cells was detected after transfection with CREB1 siRNA for 24h. KYSE-150 cells were co-transfected with anti-miR-124 and siCREB1 to exam the role of miR-124/CREB1 axis in cell proliferation and invasion. ［Results］ The expression of miR-124 was decreased and the expression of CREB1 was increased in esophagus cancer tissues(P<0.001). Overexpression of miR-124 inhibited the expression of CREB1 in KYSE-150 cells(P=0.016)，while inhibition of miR-124 promoted the expression of CREB1(P<0.001). Overexpression of miR-124 inhibited the proliferation and invasion of KYSE-150 cells(all P<0.05)，while knockdown of miR-124 inhibited the proliferation and invasion of KYSE-150 cells(all P<0.05). Knockdown of CREB1 inhibited the promoting effect of anti-miR-124 on the proliferation(P<0.05) and invasion(P<0.001) of esophageal cancer cells. ［Conclusion］ Overexpression of miR-124 inhibits prolife-ration and invasion of esophageal cancer cells by targeting CREB1. |
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