[1]鲁保才,李　靖,郜庆祖,等.MicroRNA-124对喉鳞状细胞癌细胞自噬、侵袭和迁移的影响[J].新乡医学院学报,2019,36(11):1030-1035.[doi:10.7683/xxyxyxb.2019.11.006]
　LU Bao-cai,LI Jing,GAO Qing-zu,et al.Effect of microRNA-124 on autophagy, invasion and migration of laryngeal squamous cell carcinoma cells[J].Journal of Xinxiang Medical University,2019,36(11):1030-1035.[doi:10.7683/xxyxyxb.2019.11.006]
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MicroRNA-124对喉鳞状细胞癌细胞自噬、侵袭和迁移的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年11
页码:
1030-1035
栏目:
基础研究
出版日期:
2019-11-05

文章信息/Info

Title:
Effect of microRNA-124 on autophagy, invasion and migration of laryngeal squamous cell carcinoma cells
作者:
鲁保才1; 李　靖1; 郜庆祖2; 余文发1; 卢振民1; 连　荣1; 苏　蔚2
（1.新乡医学院第一附属医院耳鼻喉科，河南　卫辉　453100；2.新乡医学院第一附属医院病理科，河南　卫辉　453100）
Author(s):
LU Bao-cai1; LI Jing1; GAO Qing-zu2; YU Wen-fa1; LU Zhen-min1; LIAN Rong1; SU Wei2
(1.Department of Otorhinolaryngology，the First Affiliated Hospital of Xinxiang Medical University，Weihui 453100，Henan Province,China；2.Department of Pathology，the First Affiliated Hospital of Xinxiang Medical University，Weihui 453100，Henan Province，China)
关键词:
microRNA-124; 喉鳞状细胞癌; 自噬; 侵袭; 迁移
Keywords:
microRNA-124; laryngeal squamous cell carcinoma; autophagy; invasion; migration
分类号:
R739.65
DOI:
10.7683/xxyxyxb.2019.11.006
文献标志码:
A
摘要:
目的　探讨microRNA-124（miR-124）在人喉鳞状细胞癌（LSCC）组织中的表达及其对LSCC细胞自噬、侵袭和迁移的影响。方法　选取2016年1月至2018年9月新乡医学院第一附属医院手术切除的LSCC组织及癌旁组织（距离肿瘤边缘1~ 2 cm）标本各12例，采用实时定量聚合酶链反应（qRT-PCR）检测LSCC组织和癌旁组织中miR-124表达；体外培养人Hep-2细胞，将其分为miR-124 类似物（miR-124 mimics）组、miR-124抑制剂（miR-124 inhibitor）组、类似物阴性对照（mimics NC）组、抑制剂阴性对照（inhibitor NC）组、mimics+西罗莫司组和inhibitor+3-甲基腺嘌呤（3-MA）组，进行细胞转染；采用qRT-PCR检测miR-124 mimics组、miR-124 inhibitor组、mimics NC组、inhibitor NC组细胞中miR-124的表达水平，Western blot法检测LSCC组织、癌旁组织和mimics NC组、miR-124 mimics组、inhibitor NC组、miR-124 inhibitor组细胞中自噬相关蛋白LC3B-Ⅱ的表达，Transwell小室法检测miR-124 mimics组、miR-124 inhibitor组、mimics NC组、inhibitor NC组、mimics+西罗莫司组和inhibitor+3-MA组细胞的侵袭能力，细胞划痕实验检测miR-124 mimics组、miR-124 inhibitor组、mimics NC组、inhibitor NC组、mimics+西罗莫司组和inhibitor+3-MA组细胞的迁移能力。结果　LSCC组织中miR-124相对表达量显著低于癌旁组织（P<0.05）。LSCC组织中LC3B-Ⅱ蛋白表达显著高于癌旁组织（P<0.05）。miR-124 mimics组细胞中miR-124相对表达量显著高于mimics NC组（P<0.05），miR-124 inhibitor组细胞中miR-124相对表达量显著低于inhibitor NC组（P<0.05）。miR-124 mimics组细胞LC3B-Ⅱ蛋白相对表达量显著低于mimics NC组（P<0.05），miR-124 inhibitor组细胞LC3B-Ⅱ蛋白相对表达量显著高于inhibitor NC组（P<0.05）。miR-124 mimics组侵袭细胞数显著少于mimics NC组和mimics+西罗莫司组（P<0.05），miR-124 inhibitor组侵袭细胞数显著多于inhibitor NC组和inhibitor+3-MA组（P<0.05）。划痕培养72 h后，inhibitor NC组和inhibitor+3-MA组细胞的划痕愈合率低于miR-124 inhibitor组（P<0.05）；培养96 h后，mimics NC组和mimics+西罗莫司组细胞的划痕愈合率高于miR-124 mimics组（P<0.05）。结论　人LSCC组织中miR-124表达显著下调，人LSCC组织和miR-124过表达细胞中自噬相关蛋白LC3B-Ⅱ表达显著上调，miR-124可能通过抑制自噬而抑制LSCC细胞的侵袭和迁移能力。
Abstract:
Objective　 To investigate the expression of microRNA-124 (miR-124) in human laryngeal squamous cell carcinoma (LSCC) and its effect on autophagy, invasion and migration of LSCC cells.Methods　Twelve LSCC tissue specimens and twelve paracancerous tissue specimens (1-2 cm away from the edge of the tumor) were collected from the First Affiliated Hospital of Xinxiang Medical University from January 2016 to September 2018.The expression of miR-124 in LSCC and paracancerous tissues was detected by real-time quantitative polymerase chain reaction(qRT-PCR)．Human LSCC Hep-2 cells were cultured in vitro and divided into miR-124 mimics group，miR-124 inhibitor group，mimics negative control(mimics NC)group,inhibitor negative control (inhibitor NC)group，mimics+sirolimus group and inhibitor+3-methyladenine(3-MA) group． After cell transfection，the expression of miR-124 in Hep-2 cells of in the miR-124 mimics group，miR-124 inhibitor group，mimics NC group and inhibitor NC group was detected by qRT-PCR.The expression of autophagy-related protein LC3B-Ⅱ in LSCC tissues,paracancerous tissues and cells in the miR-124 mimics group，miR-124 inhibitor group，mimics NC group and inhibitor NC group were detected by Western blot method.The cell invasion ability and cell migration ability in the miR-124 mimics group,miR-124 inhibitor group,mimics NC group,inhibitor NC group,mimics+sirolimus group and inhibitor+3-MA group was detected by Transwell method and would healing assay,respectively.Results　The relative expression of miR-124 in LSCC tissues was significantly lower than that in paracancerous tissues (P<0.05).The relative expression of autophagy-related protein LC3B-Ⅱ in LSCC tissues was obviously higher than that in paracancerous tissues(P<0.05).The relative expression of miR-124 in the miR-124 mimics group was significantly higher than that in the mimics NC group (P< 0.05).The relative expression of miR-124 in the miR-124 inhibitor group was significantly lower than that in the inhibitor NC group (P<0.05).The relative expression of LC3B-Ⅱ protein in the miR-124 mimics group was obviously lower than that in the mimics NC group (P<0.05).The relative expression of LC3B-Ⅱ protein in the miR-124 inhibitor group was significantly higher than that in the inhibitor NC group(P<0.05).The number of invasion cells in the miR-124 mimics group were significantly fewer than that in the mimics NC group and mimics+sirolimus group (P<0.05).The number of invasion cells in the miR-124 inhibitor group was significantly more than that in the inhibitor NC group and inhibitor+3-MA group (P<0.05).The healing rate in the inhibitor NC group and inhibitor+3-MA group was significantly lower than that in the miR-124 inhibitor group at 72 hours after scratch culturing(P<0.05).The healing rate in the mimics NC and mimics+ sirolimus group was significantly higher than that in the miR-124 mimics group at 96 hours after scratch culturing (P<0.05).Conclusion　The expression of miR-124 is significantly down-regulated in LSCC tissues.The expression of autophagy-related protein LC3B-Ⅱ is obviously up-regulated in LSCC tissues and miR-124-overexpressed cells.MiR-124 may inhibit the invasion and migration ability of LSCC cells by inhibiting autophagy.

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更新日期/Last Update: 2019-11-05