【摘要】 背景:前期研究发现,Notch2在舌鳞癌干细胞ALDH^br中表达显著上调,但其具体的作用尚不明确。目的:分析Notch2对舌鳞癌干细胞ALDH^br增殖、侵袭和迁移的影响。方法:采用无血清培养基培养舌鳞癌细胞株Tca8113,富集ALDH^br细胞,以流式细胞技术对其进行分选,采用免疫印迹检测Notch2在普通舌鳞癌肿瘤细胞ALDHlow和ALDH^br细胞中的表达水平。通过脂质体转染Notch2 sh RNA（sh-Notch2）构建Notch2沉默的ALDH^br细胞（实验组）,以转染sh-control的ALDH^br细胞为对照,48 h后检测ALDH^br细胞的增殖及Ki67蛋白表达;Transwell小室法检测两组细胞生物侵袭、迁移及基质金属蛋白酶2和基质金属蛋白酶9蛋白的表达。结果与结论:（1）Notch2在ALDH^br细胞中的表达水平显著高于其在ALDHlow细胞中的表达水平（P<0.05）;（2）实验组ALDH^br细胞的增殖能力、Ki67蛋白表达低于对照组（P<0.05）;（3）实验组ALDH^br细胞的侵袭和迁移能力显著低于对照组（P<0.05）,基质金属蛋白酶2和基质金属蛋白酶9的表达水平显著低于对照组（P<0.05）;（4）结果表明,Notch2可促进ALDH^br细胞的增殖、侵袭和迁移,提示Notch2在舌鳞癌的发生与发展中起着重要作用,其可能成为治疗舌鳞癌的分子靶点。更多还原

【Abstract】 BACKGROUND: Previous studies have found that the expression of Notch2 is up-regulated in tongue squamous cell carcinoma（TSCC） stem cells, but the effect of Notch2 on TSCC remains unclear. OBJECTIVE: To explore the effect of Notch2 on the proliferation, invasion and migration of TSCC stem cells(ALDH^br). METHODS: ALDH^br cells were enriched and separated by fluorescence-activated cell sorting technique from Tca8113 cells cultured in using serum-free medium, and Notch2 expression in ordinary TSCC cells（ALDHlow） and ALDH^br cells was detected by western blot. The ALDH^br cells transfected with Notch2 sh RNA（sh-Notch2） by liposome served as experimental group, and ALDH^br cells transfected with sh-control were as control group. The effects of Notch2 on the proliferation, invasion and migration of ALDH^br cells were detected by cell counting kit-8 kit and Transwell assay, respectively. In addition, the expression levels of Ki67, matrix metalloproteinases 2 and 9 protein in Notch2-silenced ALDH^br cells and control cells were detected by western blot. RESULTS AND CONCLUSION: The expression level of Notch2 in ALDH^br cells was significantly higher than that in ALDHlow cells（P < 0.05）. The ALDH^br cell proliferation ability and Ki67 protein expression in the experimental group were obviously lower than those in the control group（P < 0.05）. The ALDH^br cell invasion and migration abilities in the experimental group were significantly lower than those in the control group（P < 0.05）. The expression levels of matrix metalloproteinases 2 and 9 protein in the experimental group were obviously lower than those in the control group（P < 0.05）. All these results show that Notch2 can promote the proliferation, invasion and migration of ALDH^br cells, which suggests that Notch2 plays an important role in the occurrence and development of TSCC, and it may be a potential molecular target for the treatment of TSCC.更多还原