【摘 要】
：
目的：miR-330作为肿瘤抑制因子可靶向调控E2F1和Sp1表达,促进前列腺癌细胞凋亡、抑制细胞运动能力.EREG(epiregulin)是EGF成员参与并促进唾液腺腺样囊性癌(SACC)细胞肺转移及结直肠癌肝转移.miR330与EREG与肿瘤淋巴道转移相关研究未见报道.鼠肝癌低淋巴道转移力细胞株Hca-P,能够引发原位瘤及腹水瘤,淋巴结转移率～25％,是研究肿瘤淋巴道起始转移理想模型.本文旨在
【机 构】
：
大连医科大学生物技术系 大连医科大学生化教研室,大连,116044
【出 处】
：
中国生物化学与分子生物学会2016年全国学术会议
论文部分内容阅读
目的：miR-330作为肿瘤抑制因子可靶向调控E2F1和Sp1表达,促进前列腺癌细胞凋亡、抑制细胞运动能力.EREG(epiregulin)是EGF成员参与并促进唾液腺腺样囊性癌(SACC)细胞肺转移及结直肠癌肝转移.miR330与EREG与肿瘤淋巴道转移相关研究未见报道.鼠肝癌低淋巴道转移力细胞株Hca-P,能够引发原位瘤及腹水瘤,淋巴结转移率～25％,是研究肿瘤淋巴道起始转移理想模型.本文旨在建立mmu-miR-330通过EREG调控Hca-P恶性表型的作用途径及机制.方法：生物芯片结果发现mmu-miR-330与鼠肝癌淋巴道转移相关；用阳离子脂质体法将mmu-miR-330 inhibitor转染入Hca-P,qRT-PCR法检测mmu-miR-330下调效率；Transwell法检测mmu-miR-330下调对Hca-P迁移及侵袭能力影响；CCK-8法检测mmu-miR-330下调对Hca-P体外增殖能力影响；运用TargetScan,miRDB等软件分析EREG与mmu-miR-330靶向关系；双荧光素酶报告基因实验检测在Hca-P中上调mmu-miR-330表达对EREG的影响；qRTPCR法检测Vimentin、MMP9和E-cadherin在Hca-P中随mmu-miR-330变化表达水平变化.结果：高淋巴道转移力(75％)Hca-F相比,mmu-miR-330在Hca-P中水平升高200％；下调mmu-miR-330表达促进Hca-P的迁移和侵袭能力,对其增殖能力无明显影响；生物信息学表明,mmu-miR-330可能通过EREG 3UTR区的两个结合位点结合调控其表达；双荧光素酶报告基因实验显示,mmu-miR-330与EREG 3UTR中701-724区域靶点结合明显,但与492-512区域的靶点结合不明显；mmu-miR-330下调促进VIMENTIN和MMP9,降低E-cadherin在Hca-P中表达.结论：Mmu-miR-330下调增强Hca-P体外迁移侵袭能力,不影响其增殖.Mmu-miR-330下调使EREG表达水平升高,二者呈现负靶向调控,mmu-miR-330对与EREG 3UTR中701-724区域结合对其起抑制作用.mmu-miR-330通过改变E-cadherin,Vimentin和MMP9的表达调控Hca-P的恶性表型.
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