中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2014年
12期
990-994
,共5页
洒娜%于亮%牟亚魁%孙高英%徐伟
灑娜%于亮%牟亞魁%孫高英%徐偉
쇄나%우량%모아괴%손고영%서위
下咽肿瘤%微RNAs%Twist转录因子%肿瘤侵润
下嚥腫瘤%微RNAs%Twist轉錄因子%腫瘤侵潤
하인종류%미RNAs%Twist전록인자%종류침윤
Hypopharyngeal neoplasms%microRNAs%Twist transcription factor%Neoplasm invasiveness
目的 研究microRNA-214(miR-214)在晚期下咽鳞状细胞癌(鳞癌)组织中的表达情况,及其对下咽癌FaDu细胞系侵袭、迁移、克隆形成能力的影响,并分析其可能机制.方法 利用qRT-PCR技术检测30例晚期下咽鳞癌患者肿瘤组织及正常黏膜组织中miR-214的表达;通过转染过表达载体hsa-mir-214上调FaDu细胞中miR-214的表达;采用Transwell侵袭、迁移实验和平板克隆实验,分别检测miR-214表达上调后对FaDu细胞侵袭、迁移及克隆形成能力的影响;利用Western blot检测上调miR-214表达对Twist蛋白表达的影响.结果 miR-214在晚期下咽鳞癌组织中相对表达((x)±s)为(0.311±0.206)明显低于下咽正常黏膜组织中的相对表达为(1.620±1.394),差异有统计学意义(=5.09,P＜0.05).实验组FaDu细胞转染hsa-mir-214后,miR-214的表达较阴性对照组明显上调(t=6.347,P＜0.05).Transwell迁移和侵袭实验显示,实验组FaDu细胞迁移能力及侵袭能力均明显低于对照组(t=11.6,P＜0.01;t=6.499,P＜0.05).平板克隆实验显示,实验组与对照组相比,平均克隆形成数及克隆形成率差异均无统计学意义(t=0.592,P＞0.05),miR-214对FaDu细胞增殖能力无明显影响.Western blot结果显示,miR-214上调后的实验组细胞Twist蛋白的表达明显低于对照组细胞中的表达(t=6.545,P＜0.05).结论 miR-214在晚期下咽鳞癌组织中低表达,且可明显抑制FaDu细胞的侵袭、迁移能力,其作用机制可能与抑制Twist蛋白表达有关.miR-214对FaDu细胞增殖能力无明显影响.
目的 研究microRNA-214(miR-214)在晚期下嚥鱗狀細胞癌(鱗癌)組織中的錶達情況,及其對下嚥癌FaDu細胞繫侵襲、遷移、剋隆形成能力的影響,併分析其可能機製.方法 利用qRT-PCR技術檢測30例晚期下嚥鱗癌患者腫瘤組織及正常黏膜組織中miR-214的錶達;通過轉染過錶達載體hsa-mir-214上調FaDu細胞中miR-214的錶達;採用Transwell侵襲、遷移實驗和平闆剋隆實驗,分彆檢測miR-214錶達上調後對FaDu細胞侵襲、遷移及剋隆形成能力的影響;利用Western blot檢測上調miR-214錶達對Twist蛋白錶達的影響.結果 miR-214在晚期下嚥鱗癌組織中相對錶達((x)±s)為(0.311±0.206)明顯低于下嚥正常黏膜組織中的相對錶達為(1.620±1.394),差異有統計學意義(=5.09,P＜0.05).實驗組FaDu細胞轉染hsa-mir-214後,miR-214的錶達較陰性對照組明顯上調(t=6.347,P＜0.05).Transwell遷移和侵襲實驗顯示,實驗組FaDu細胞遷移能力及侵襲能力均明顯低于對照組(t=11.6,P＜0.01;t=6.499,P＜0.05).平闆剋隆實驗顯示,實驗組與對照組相比,平均剋隆形成數及剋隆形成率差異均無統計學意義(t=0.592,P＞0.05),miR-214對FaDu細胞增殖能力無明顯影響.Western blot結果顯示,miR-214上調後的實驗組細胞Twist蛋白的錶達明顯低于對照組細胞中的錶達(t=6.545,P＜0.05).結論 miR-214在晚期下嚥鱗癌組織中低錶達,且可明顯抑製FaDu細胞的侵襲、遷移能力,其作用機製可能與抑製Twist蛋白錶達有關.miR-214對FaDu細胞增殖能力無明顯影響.
목적 연구microRNA-214(miR-214)재만기하인린상세포암(린암)조직중적표체정황,급기대하인암FaDu세포계침습、천이、극륭형성능력적영향,병분석기가능궤제.방법 이용qRT-PCR기술검측30례만기하인린암환자종류조직급정상점막조직중miR-214적표체;통과전염과표체재체hsa-mir-214상조FaDu세포중miR-214적표체;채용Transwell침습、천이실험화평판극륭실험,분별검측miR-214표체상조후대FaDu세포침습、천이급극륭형성능력적영향;이용Western blot검측상조miR-214표체대Twist단백표체적영향.결과 miR-214재만기하인린암조직중상대표체((x)±s)위(0.311±0.206)명현저우하인정상점막조직중적상대표체위(1.620±1.394),차이유통계학의의(=5.09,P＜0.05).실험조FaDu세포전염hsa-mir-214후,miR-214적표체교음성대조조명현상조(t=6.347,P＜0.05).Transwell천이화침습실험현시,실험조FaDu세포천이능력급침습능력균명현저우대조조(t=11.6,P＜0.01;t=6.499,P＜0.05).평판극륭실험현시,실험조여대조조상비,평균극륭형성수급극륭형성솔차이균무통계학의의(t=0.592,P＞0.05),miR-214대FaDu세포증식능력무명현영향.Western blot결과현시,miR-214상조후적실험조세포Twist단백적표체명현저우대조조세포중적표체(t=6.545,P＜0.05).결론 miR-214재만기하인린암조직중저표체,차가명현억제FaDu세포적침습、천이능력,기작용궤제가능여억제Twist단백표체유관.miR-214대FaDu세포증식능력무명현영향.
Objective To investigate the expression of microRNA-214 (miR-214) in advanced hypopharyngeal carcinoma tissues and its effects on the invasion,migration and colone formation of FaDu cells.Methods miR-214 expression in 30 cases of advanced hypopharyngeal carcinoma tissues and normal hypopharyngeal mucosa tissues was detected by real-time quantitative polymerase chain reaction (RT-qPCR).miR-214 was upregulated through transfecting the overexpression vector hsa-mir-214 into FaDu cells.The influences of miR-214 upregulation on the invasion,migration,clone formation and Twist expression were measured by Transwell invasion,Transwell migration,plate clone formation and Western blot assays,respectively.Results The expression of miR-214 in advanced hypopharyngeal carcinoma tissues (0.31 1 ±0.206) was significantly less than normal hypopharyngeal mucosa tissues(1.620 ± 1.394;t =5.09,P ＜ 0.05).The expression of miR-214 was notably upregulated after tranfected with hsa-mir-214 compared with the negative control group (t =6.347,P ＜ 0.05).The migration and invasion ability of FaDu cells transfeced with hsa-mir-214 was decreased by comparison with negative control cells (t =1 1.6,P ＜0.01; t =6.499,P ＜ 0.05).There was no significant difference of the average clony number and the cloning efficiency between the experimental and negative control groups (t =0.592,P ＞ 0.05).Results of Western blot assay showed that,Twist expression in the miR-214-overexpressed group was apparently decreased compared with that in the control group (t =6.545,P ＜ 0.05).Conclusions miR-214 is expressed at a low level in advanced hypopharyngeal carcinoma tissues,and can obviously inhibit the invasion and migration abilities of FaDu cells,possibly because of its inhibiting effect on Twist expression.Additionally,miR-214 plays no significant role in the proliferation of FaDu cells.
