华西口腔医学杂志 ›› 2020, Vol. 38 ›› Issue (2): 139-144.doi: 10.7518/hxkq.2020.02.005

• 基础研究 • 上一篇    下一篇

沉默热休克蛋白27基因对头颈部鳞状细胞癌细胞生物学行为的影响

朱震坤, 王羽裳, 徐欣()   

  1. 山东大学口腔医学院·口腔医院种植科 山东省口腔组织再生重点实验室 山东省口腔生物材料与组织再生工程实验室,济南 250012
  • 收稿日期:2019-08-12 修回日期:2020-01-13 出版日期:2020-04-01 发布日期:2020-04-15
  • 通讯作者: 徐欣 E-mail:xinxu@sdu.edu.cn
  • 作者简介:朱震坤,讲师,博士,E-mail:zhenkunzhu@sdu.edu.cn
  • 基金资助:
    泰山学者建设项目专项基金(ts201511106);山东大学交叉学科项目(2016JC024)

Biology behavior of head and neck squamous cell cancer cells changes after knocking down heat shock protein 27

Zhu Zhenkun, Wang Yushang, Xu Xin()   

  1. Dept. of Implantology, School and Hospital of Stomatology, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan 250012, China
  • Received:2019-08-12 Revised:2020-01-13 Online:2020-04-01 Published:2020-04-15
  • Contact: Xin Xu E-mail:xinxu@sdu.edu.cn
  • Supported by:
    Shandong Taishan Scholar Project Special Fund(ts201511106);Interdisciplinary Program of Shandong University(2016JC024)

摘要:

目的 观察长效沉默热休克蛋白(Hsp)27基因后头颈部鳞状细胞癌细胞生物学行为的改变。方法 实验分为3组:高滴度pLenti-shRNA-Hsp27慢病毒颗粒长效转染入UM-SCC-22B细胞为实验组(shHsp27组),常规培养UM-SCC-22B细胞(ctrl组)为空白对照,UM-SCC-22B细胞转染pLenti-shRNA-ctrl慢病毒颗粒(shctrl组)为阴性对照。采用实时荧光定量聚合酶链反应和蛋白质印迹法检测各组中Hsp27的表达,采用MTS细胞增殖实验、细胞划痕实验及Matrigel侵袭实验,观察各组Hsp27表达抑制后UM-SCC-22B细胞的增殖、迁移和侵袭能力的变化。结果 shHsp27组的Hsp27表达明显降低;MTS细胞增殖实验可见,细胞培养24、48 h后,shHsp27组的细胞增殖能力与ctrl组和shctrl组无明显差异;划痕实验表明,划痕产生72 h后,ctrl组细胞迁移能力为shHsp27组的4.38倍;Matrigel侵袭实验显示,ctrl组细胞的体外侵袭能力为shHsp27组的2.03倍。结论 长效转染慢病毒颗粒pLenti-shRNA-Hsp27能够高效、特异地沉默高转移潜能头颈部鳞状细胞癌细胞系UM-SCC-22B的Hsp27基因表达,并能显著抑制其体外侵袭和转移能力。

关键词: 热休克蛋白27, 人头颈部鳞状细胞癌, 转移, 慢病毒转染

Abstract:

Objective This study aimed to observe the metastatic behavior of head and neck squamous cell carcinoma cells after knocking down heat shock protein (Hsp) 27. Methods The experiment was divided into three groups: the lentivirus vector plasmid of pLenti-shRNA-Hsp27 was transfected into UM-SCC-22B cells as experimental group (shHsp27 group), routine culture of UM-SCC-22B cells as blank control (ctrl group), UM-SCC-22B cells transfection of pLenti-shRNA-ctrl lentivirus vector as negative control (shctrl group). Through real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot assay to detect the mRNA expression of Hsp27 in three groups. MTS assay was performed to detect cell-proliferation changes, wounding healing assay was performed to detect cell-migration changes, and Matrigel Transwell invasion assay was performed to detect cell-invasion changes. Results The expression of Hsp27 in shHsp27 group decreased significantly; MTS assay showed that UM-SCC-22B before and after Hsp27 knockdown had similar proliferation rates after being cultured for 24 or 48 h. Compared with the ctrl group, the shHsp27 group decreased the metastatic behavior by 4.38-fold in migration and 2.03-fold in cell invasion. Conclusion Stably transfected lentivirus vector plasmid of pLenti-shRNA-Hsp27 can efficiently decrease Hsp27 expression and reduce the metastasis ability of UM-SCC-22B.

Key words: heat shock protein 27, human head and neck squamous cell cancer, metastasis, lentivirus transfection

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