过表达miR-141对舌癌Tscca和Tca-8113细胞侵袭、迁移的影响  

Effect of overexpression of miR-141 in Tscca and Tca-8113 on cell invasion and migration

作　　者：黄涛 刘慧 邵博[1] 龚忠诚[1] HUANG Tao;LIU Hui;SHAO Bo;GONG Zhongcheng(Department of Oral and Maxillofacial Tumor Surgery,the First Affiliated Hospital of Xinjiang Medical University/Affiliated Stomatological Hospital,Xinjiang Urumqi 830054,China;Department of Oral and Maxillofacial Surgery,Shanghai Stomatological Hospital,Fudan University,Shanghai 200001,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)颌面肿瘤外科,新疆维吾尔自治区口腔医学研究所,新疆乌鲁木齐830054 [2]上海市口腔疾病防治医院颌面外科,上海200001

出　　处：《现代肿瘤医学》2021年第7期1097-1102,共6页Journal of Modern Oncology

基　　金：国家自然科学基金(编号:81460439)。

摘　　要：目的:研究过表达miRNA-141对舌癌细胞迁移和侵袭的影响。方法:使用miR-141慢病毒Ubi-miR-141-SV40-EGFP-IRES-puromycin(miR-141组)和慢病毒空载体Ubi-MCS-SV40-EGFP-IRES-puromycin(miR-NC组)转染两组舌癌细胞系(Tscca、Tca-8113细胞系),未转染的细胞为空白组(Blank)。采用荧光定量PCR(RT-qPCR)检测miR-141基因表达,RT-qPCR和蛋白免疫印迹(Western blot)检测基质金属蛋白酶2、9(MMP2、9),E盒结合锌指蛋白2(zeb2)的mRNA和蛋白表达。结果:流式细胞术测定转染率,Tscca细胞中miR-141组转染率为82.4%、miR-NC组为87.3%;Tca-8113细胞中miR-141组转染率为73.4%、miR-NC组为89.1%。RT-qPCR结果显示:miR-141组中miR-141表达明显高于miR-NC组和Blank组,证明稳定转染的细胞株建立成功。Tscca和Tca-8113细胞中miR-141组和miR-NC组、Blank组组间比较,MMP2、MMP9、zeb2的mRNA和蛋白表达均降低。结论:在Tscca和Tca-8113细胞中通过过表达miR-141抑制细胞中MMP2、MMP9、zeb2的mRNA和蛋白表达来控制细胞迁移和侵袭能力。Objective:To detect the effects of miR-141 on invasion and migration of tongue cancer cell(Tscca,Tca-8113).Methods:Both groups of tongue cancer cells were stably transfected with miR-141 lentivirus Ubi-miR-141-SV40-EGFP-IRES-puromycin(miR-141 group)and lentiviral empty vector Ubi-MCS-SV40-EGFP-IRES-puromycin(miR-NC group).Untreated cells were blank group(Blank).Reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-141.Flow cytometry was used to measure transfection efficiency.Wound-healing analysis,migration and invasion were performed to detect cell functions.The mRNA and protein expression of MMP2/9(matrix metalloprotein 2,MMP2,matrix metalloprotein 9,MMP9)and zinc finger E-box binding homeobox 2(zeb2)were detected by RT-qPCR and Western blot.Results:The GFP transfection rate was determined by flow cytometry.In Tscca cells,the transfection rate of miR-141 group was 82.4%and 87.3%in the miR-NC group.In Tca-8113 cells,the transfection rate of miR-141 group was 73.4%and 89.1%in the miR-NC group.RT-qPCR showed that the expression of miR-141 in miR-141 group was significantly higher than that of miR-NC group and Blank group,which proved that the stable transfected cell line was successfully established.The mRNA and protein expressions of MMP2,MMP9 and zeb2 were decreased in miR-141 group compared with miR-NC group and Blank group.Conclusion:In Tscca and Tca-8113 cells,overexpression of miR-141 inhibits tongue cancer mRNA and protein expression of MMP2,MMP9 and zeb2 to control cell invasion and migration.

关 键 词：舌癌 microRNA-141 基质金属蛋白 E盒结合锌指蛋白2 上皮间质转化 

分 类 号：R739.86[医药卫生—肿瘤]