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
Biology behavior of head and neck squamous cell cancer cells changes after knocking down heat shock protein 27
作者：朱震坤, 王羽裳, 徐欣
Author：Zhu Zhenkun, Wang Yushang, Xu Xin
收稿日期：2019-08-12 年卷（期）页码：2020,38(2):139-139-144
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：热休克蛋白27,人头颈部鳞状细胞癌,转移,慢病毒转染,
Key words：heat shock protein 27,human head and neck squamous cell cancer,metastasis,lentivirus transfection,
基金项目：泰山学者建设项目专项基金(ts201511106);山东大学交叉学科项目(2016JC024)
中文摘要
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。
英文摘要
ObjectiveThis study aimed to observe the metastatic behavior of head and neck squamous cell carcinoma cells after knocking down heat shock protein (Hsp) 27.MethodsThe 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.ResultsThe 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.ConclusionStably transfected lentivirus vector plasmid of pLenti-shRNA-Hsp27 can efficiently decrease Hsp27 expression and reduce the metastasis ability of UM-SCC-22B.
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