miR-302c调控CXCL8表达抑制食管鳞状细胞癌细胞的增殖和侵袭

龚向南; 苏静; 陈玮煦; 李祥全

doi:10.13898/j.cnki.issn.1000-2200.2023.12.002

miR-302c调控CXCL8表达抑制食管鳞状细胞癌细胞的增殖和侵袭

MiR-302c regulates the expression of CXCL8 to inhibit the proliferation and invasion of esophageal squamous cell carcinoma cells

摘要

摘要:
目的探讨miR-302c调控CXC趋化因子配体8（CXCL8）表达对食管鳞状细胞癌（ESCC）细胞增殖和侵袭的影响。
方法收集行手术治疗的ESCC病人ESCC组织和其对应的癌旁组织，以及人ESCC细胞系Eca109、Ec9706、TE-11、TE-10、食管正常上皮细胞系Het-1A为研究对象，qRT-PCR检测miR-302c表达；利用Lipofectamine^TM2000转染试剂盒对Eca109细胞进行转染，分组为：空白组（细胞未转染）、miR-NC组、miR-302c mimics组、si-CXCL8组、si-NC组、miR-302c mimics+OE-NC组、miR-302c mimics+OE-CXCL8组，qRT-PCR检测各组细胞中miR-302c表达；MTT法检测各组细胞增殖情况；Transwell实验检测各组细胞侵袭情况；双荧光素酶报告基因检测实验验证miR-302c与CXCL8靶向关系；Western blotting检测CXCL8、细胞周期蛋白D1（Cyclin D1）、基质金属蛋白酶（MMP）-2、MMP-9蛋白表达情况。
结果miR-302c在ESCC组织中的表达水平低于癌旁组织（P < 0.05），与人食管正常上皮细胞系Het-1A比较，人ESCC细胞系Eca109、Ec9706、TE-11、TE-10中miR-302c表达水平降低（P < 0.05），且Eca109细胞中miR-302c表达水平最低，因此，选择Eca109细胞进行后续研究；miR-302c靶向负调控CXCL8表达；上调miR-302c和沉默CXCL8均能抑制Eca109细胞的增殖与侵袭，并降低Cyclin D1、MMP-2、MMP-9蛋白表达（P < 0.05）；过表达CXCL8可逆转上调miR-302c对Eca109细胞增殖与侵袭的影响。
结论上调miR-302c表达可通过靶向抑制CXCL8表达，抑制ESCC细胞的增殖和侵袭。

Abstract:
ObjectiveTo investigate the effects of miR-302c on regulating the expression of CXC chemokine ligand 8 (CXCL8) on the proliferation and invasion of esophageal squamous cell carcinoma (ESCC) cells.
MethodsThe ESCC tissues and their corresponding adjacent tissues were collected from ESCC patients who underwent surgical treatment, as well as human ESCC cell lines Eca109, Ec9706, TE-11, TE-10 and normal esophageal epithelial cell line Het-1A were set as the study subjects, the expression of miR-302c was detected by qRT-PCR.Eca109 cells were transfected with Lipofectamine^TM2000 transfection kit and divided into blank group (cells not transfected), miR-NC group, miR-302c mimics group, si-CXCL8 group, si-NC group, miR-302c mimics +OE-NC group and miR-302c mimics+OE-CXCL8 group, the expression of miR-302c in each group was detected by qRT-PCR; the cell proliferation in each group was detected by MTT method; the cell invasion in each group was detected by Transwell assay; the dual luciferase reporter gene detection experiment was used to verify the targeting relationship between miR-302c and CXCL8;and the protein expressions of CXCL8, Cyclin D1, matrix metalloproteinase (MMP)-2 and MMP-9 were measured by Western blotting.
ResultsThe expression level of miR-302c in ESCC tissues was significantly lower than that in adjacent tissues (P < 0.05), when compared with the normal human esophageal epithelial cell line Het-1A, the expression level of miR-302c in human ESCC cell lines Eca109, Ec9706, TE-11, TE-10 was significantly reduced (P < 0.05), and the expression level of miR-302c was the lowest in Eca109 cells, therefore, Eca109 cells were selected for follow-up research.miR-302c targeted and negatively regulated the expression of CXCL8;up-regulating miR-302c and silencing CXCL8 could both inhibit the proliferation and invasion of Eca109 cells, and significantly reduce the protein expressions of Cyclin D1, MMP-2 and MMP-9 (P < 0.05);overexpression of CXCL8 could reverse the effect of up-regulation of miR-302c on the proliferation and invasion of Eca109 cells.
ConclusionsUp-regulating the expression of miR-302c can inhibit the proliferation and invasion of ESCC cells by targeting and inhibiting the expression of CXCL8.

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