摘要
目的探讨微小RNA-10b-3p (miR-10b-3p)对食管鳞癌(ESCC)细胞增殖、迁移和侵袭能力的影响。方法采用QPCR检测miR-10b-3p在ESCC TE-1、EC109、KYSE30、KYSE150、KYSE180、KYSE450和KYSE510细胞株中的相对表达量,选取相对表达水平最高和最低的细胞株分别转染miR-10b-3p抑制剂和模拟物,并设转染对照载体的对照组。采用MTS实验、克隆集落形成实验及Transwell趋化小室实验检测miR-10b-3p对ESCC细胞增殖,克隆形成及侵袭、迁移能力的影响。结果 miR-10b-3p在TE-1、EC109、KYSE30、KYSE150、KYSE180、KYSE450以及KYSE510细胞株中的相对表达量分别为0. 14、0. 27、0. 66、0. 10、0. 12、0. 08和0. 78。选取miR-10b-3p相对表达量最低的KYSE450细胞株和相对表达量最高的KYSE510细胞进行后续实验。MTS检测显示,与对照组比较,模拟物组KYSE450细胞的增殖能力显著上调(P<0. 05),抑制剂组KYSE510细胞的增殖能力显著降低(P<0. 05)。克隆集落形成实验显示,模拟物组KYSE450细胞的集落数为(613. 7±41. 1)个,高于对照组的(243. 7±25. 3)个(P <0. 05);抑制剂组KYSE510细胞的集落数为(90. 3±11. 0)个,低于对照组的(247. 0±14. 7)个(P<0. 05)。Transwell小室实验显示,模拟物组穿过迁移小室的KYSE450细胞数为(211. 1±27. 3)个,高于对照组的(151. 7±15. 3)个(P<0. 05);模拟物组穿过侵袭小室的细胞数为(183. 3±15. 5)个,高于对照组的(133. 5±8. 8)个(P<0. 05)。抑制剂组穿过迁移小室的KYSE510细胞数为(50. 0±6. 7)个,低于对照组的(83. 3±12. 9)个(P<0. 05);抑制剂组穿过侵袭小室的KYSE510细胞数为(38. 8±8. 5)个,低于对照组的(66. 7±7. 3)个(P<0. 05)。结论 miR-10b-3p可以促进ESCC细胞的增殖、侵袭和迁移,可能作为ESCC基因治疗的有效靶点。
Objective To explore the expression of microRNA-10b-3p(miR-10b-3p)in esophageal squamous cell carcinoma(ESCC)and its effect on proliferation,migration and invasion of ESCC cell lines.Methods The relative expression of miR-10b-3p in TE-1,EC109,KYSE30,KYSE150,KYSE180,KYSE450and KYSE510cell lines was detected by QPCR.The cell lines with highest and lowest expression of miR-10b-3p were transfected with miR-10b-3p mimics and inhibitor respectively,and meanwhile,control vectors were set as control.Furthermore,cell proliferation,colony formation,migration and invasion were respectively analyzed by MTS assay,colon formation experiment and Transwell chamber experiment.Results The relative miR-10b-3p expression in TE-1,EC109,KYSE30,KYSE150,KYSE180,KYSE450and KYSE510cell lines was0.14,0.27,0.66,0.10,0.12,0.08and0.78.The results showed that miR-10b-3p was expressed lower in KYSE450cell line,but higher in KYSE510cell line,and they were chosen for the following experiments.Then,miR-10b-3p mimic and inhibitor were respectively transferred into KYSE450and KYSE510cell lines.MTS assay showed that compared with control group,transfection of KYSE450cell line with miR-10b-3p mimic significantly promoted the cell viability(P<0.05),and transfection of KYSE510cell line with miR-10b-3p inhibitor could decrease the cell viability(P<0.05).What s more,the colony formation(613.7±41.1vs.243.7±25.3,P<0.05),migration(211.1±27.3vs.151.1±15.3)and invasion(183.3±15.5vs.133.5±8.8,P<0.05)ability of KYSE450cell line were also promoted as a result of miR-10b-3p mimic transfection.Lastly,down-regulation of miR-10b-3p using inhibitor could inhibite the malignant phenotype of KYSE510ESCC cells in vitro,including colony formation(90.3±11.0vs.247.0±14.7,P<0.05),as well as cell migration(50.0±6.7vs.83.3±12.9,P<0.05)and invasion(38.8±8.5vs.66.7±7.3,P<0.05).Conclusion MiR-10b-3p can increase the malignant phenotype of ESCC cells,and therefore it can be used as an effective target for gene therapy of ESCC.
作者
张超
张洁
高鹏
王欢
曹春晓
刘美月
陈思远
杨朝
缐丽歌
张立新
孙国贵
ZHANG Chao;ZHANG Jie;GAO Peng;WANG Huan;CAO Chunxiao;LIU Meiyue;CHEN Siyuan;YANG Zhao;XIAN Lige;ZHANG Lixin;SUN Guogui(Department of Radiotherapy, Tangshan People s Hospital, Tangshan 063001, China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2018年第11期961-966,共6页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(81402534)
河北省自然科学基金资助项目(H2015105095)
河北省医学科学研究重点课题计划资助项目(ZD20140084)
河北省"三三人才工程"计划资助项目(A2016002090
A201801005)
河北省青年拔尖人才计划资助项目(2016)10号
唐山市分子肿瘤研究与临床转化应用创新团队资助项目(15130234a)
