首页|miR-27a靶向调控SFRP1对结直肠癌生物学行为的影响

miR-27a靶向调控SFRP1对结直肠癌生物学行为的影响

Effect of miR-27a targeting regulation of SFRP1 on biological behavior of colorectal cancer

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目的 探讨miR-27a在结直肠癌中的表达,并分析其靶向调控分泌型卷曲相关蛋白 1(SFRP1)对人结直肠癌细胞HCT116 生物学行为的影响.方法 实时荧光定量 PCR(qRT-PCR)检测结直肠癌组织及癌旁正常组织中 miR-27a和SFRP1 mRNA的表达情况;Western blot检测结直肠癌组织及癌旁正常组织中SFRP1 蛋白表达水平;运用TargetScan预测软件及双荧光素酶报告基因实验检测 miR-27a 对SFRP1 的靶向调控作用;将 miR27a 模拟物(miR-27a mim-ic)、miR-27a抑制物(miR-27a inhibitor)及阴性对照(NC)转染至HCT116 细胞中;采用qRT-PCR测定各组细胞中miR-27a和SFRP1 mRNA的表达水平;运用四唑盐(MTT)比色法检测各组细胞增殖情况;Transwell实验评估各组细胞侵袭和迁移能力;Western blot检测各组细胞中SFRP1、Wnt/β-cate-nin信号通路中的关键因子Wnt4 和β-catenin的蛋白表达水平.结果 与癌旁正常组织相比,miR-27a在结直肠癌组织中高表达,而SFRP1 在结直肠癌组织中低表达(P<0.05);TargetScan预测软件和双荧光素酶报告基因实验表明miR-27a靶向调控SFRP1;与NC组相比,miR-27a mimic组miR-27a表达水平升高,细胞增殖、侵袭和迁移能力增强,SFRP1蛋白表达降低,Wnt4 和β-catenin蛋白表达增加(P<0.05);与miR-27a mimic 组相比,miR-27a inhibitor 组细胞内 miR-27a表达下降,细胞增殖、侵袭和迁移能力降低,SFRP1 蛋白表达增加,Wnt4 和β-catenin蛋白表达降低(P<0.05).结论 miR-27a能够靶向调控SFRP1,通过上调SFRP1,阻断下游的Wnt/β-catenin信号通路,抑制结直肠癌细胞的增殖、侵袭和迁移等生物学过程,从而发挥抑癌作用.为临床治疗提供了新方向.
Objective To investigate the expression of miR-27a in colorectal cancer cell,and to analyze the effect of its targeted regulation of(Secreted Frizzled-Related Protein,SFRP1)on the biological behavior of colorectal cancer cells.Methods Real-time fluorescent quantitative PCR(qRT-PCR)was employed to examine the expres-sion of miR-27a and SFRP1 mRNA in colorectal cancer tissues and adjacent normal tissues.Western blot was used to detect the expression of SFRP1 protein in colorectal cancer tissues and adjacent normal tissues.TargetScan soft-ware and dual luciferase reporter gene test were used to detect the targeted regulation of miR-27a on SFRP1.HCT116 cells were transfected with miR-27a mimic,miR-27a inhibitor and negtive control(NC).The expression of miR-27a and SFRP1 mRNA in each group was determined by qRT-PCR.MTT colorimetry was performed to eval-uate the proliferation of each group cells.Transwell assay was used to evaluate the cell invasion and migration abili-ty.Meanwhile,the protein expression levels of SFRP1,key factors Wnt4 and β-catenin in the Wnt/β-catenin sig-naling pathway were determined by Western blot.Results Compared with adjacent normal tissues,miR-27a was highly expressed in colorectal cancer tissues,while SFRP1 was low expressed in colorectal cancer tissues(P<0.05).TargetScan software and dual luciferase reporter gene test showed that miR-27a targeted SFRP1.Compared with NC group,the expression of miR-27a of miR-27a mimic group increased,the proliferation,invasion and mi-gration ability enhanced,the expression of SFRP1 protein decreased,while Wnt4 and β-catenin protein expression increased(P<0.05).Compared with miR-27a mimic group,the expression of miR-27a of miR-27a inhibitor group decreased,the proliferation,invasion and migration ability reduced,the expression of SFRP1 protein in-creased,while Wnt4 and β-catenin protein expression decreased(P<0.05).Conclusion miR-27a can target SFRP1,inhibit the proliferation,invasion and migration of colorectal cancer cells,mainly by up-regulating SFRP1 and blocking the downstream Wnt/β-catenin signaling pathway,which provides a new direction for clinical treat-ment.

miR-27aSFRP1colorectal cancerWnt4β-catenin

司马学琴、苏延停、曾智

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湖北科技学院医学部基础医学院 组胚教研室,咸宁 437100

湖北科技学院医学部基础医学院 生物化学与分子生物学教研室,咸宁 437100

咸宁市中心医院病理科,咸宁 437100

miR-27a SFRP1 结直肠癌 Wnt4 β-catenin

国家自然科学基金湖北科技学院校内科研发展项目湖北科技学院教学研究项目

320009062020-21X292023XK028

2024

安徽医科大学学报
安徽医科大学

安徽医科大学学报

CSTPCD北大核心
影响因子:1.095
ISSN:1000-1492
年,卷(期):2024.59(3)
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