中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (25): 3995-4000.doi: 10.3969/j.issn.2095-4344.2017.25.010

• 肿瘤干细胞 cancer stem cells • 上一篇    下一篇

特异性干扰TACC3基因表达对CD133+CD44+口腔鳞癌干细胞增殖及凋亡的影响

段 瑞,李永生,夏翼超   

  1. 云南省第一人民医院口腔颌面外科,云南省昆明市  650032
  • 修回日期:2017-04-14 出版日期:2017-09-08 发布日期:2017-10-09
  • 通讯作者: 李永生,主任医师,博士,云南省第一人民医院口腔颌面外科,云南省昆明市 650032
  • 作者简介:段瑞,女,1976年生,云南省昆明市人,硕士,主治医师,主要从事口腔颌面部肿瘤诊治方向的研究。
  • 基金资助:

    昆明医科大学应用基础联合专项基金面上项目

Effects of specific interfering TACC3 gene expression on proliferation and apoptosis of CD133+CD44+ oral squamous cell carcinoma cells

Duan Rui, Li Yong-sheng, Xia Yi-chao   

  1. Department of Oral and Maxillofacial Surgery, First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • Revised:2017-04-14 Online:2017-09-08 Published:2017-10-09
  • Contact: Li Yong-sheng, M.D., Chief physician, Department of Oral and Maxillofacial Surgery, First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • About author:Duan Rui, Master, Attending physician, Department of Oral and Maxillofacial Surgery, First People’s Hospital of Yunnan Province, Kunming 650032, Yunnan Province, China
  • Supported by:

    the Application-Basis Joint Special Fund of Kunming Medical University

摘要:

文章快速阅读:

文题释义:
TACC3:
是TACC蛋白家族成员之一,是细胞分裂过程中的重要分子。TACC3可通过调控微管蛋白质的组成,进而影响细胞的有丝分裂及染色体的稳定性。多项研究表明,在人类的乳腺和卵巢组织肿瘤中TACC3表达上调,用紫杉醇治疗宫颈癌和乳腺癌后TACC3的表达明显下调。体外特异性干预TACC3的表达,可有效抑制肿瘤细胞的生长,降低化疗耐药。
肿瘤干细胞的分离培养方法:主要有2种,一种是利用流式细胞仪或免疫磁珠对细胞表面表达特定的抗原分子等阳性细胞进行富集,得到含量较高的肿瘤干细胞;另一种方法是使用无血清培养基培养,通过非干细胞凋亡而干细胞成球的特性筛选出肿瘤干细胞。

 

摘要
背景:
研究表明,TACC3基因在多种肿瘤中均表达上调,因此体外特异性干预TACC3的表达,可能是治疗或干预肿瘤生长的重要靶点。
目的:探讨沉默TACC3基因表达对口腔鳞癌干细胞增殖和凋亡的影响。  
方法:采用免疫磁珠分选技术从人口腔鳞癌细胞Cal-27中分选出CD133+CD44+口腔鳞癌干细胞。shRNA转染组设计及合成TACC3 shRNA序列,利用LipofectamineTM2000将TACC3 shRNA序列转染至CD133+ CD44+口腔鳞癌干细胞,设置空载体转染阴性对照组和未转染组。转染48 h后,采用MTT、细胞克隆形成实验、TUNEL凋亡实验检测沉默TACC3基因对体外CD133+CD44+口腔鳞癌干细胞增殖、凋亡的影响;Western blot检测沉默TACC3基因对CD133+CD44+口腔鳞癌干细胞Ki67、Bax、Bcl-2蛋白表达的影响。
结果与结论:①细胞增殖:shRNA转染组细胞增殖速度、增殖相关蛋白Ki67表达水平明显低于阴性对照组和未转染组(P < 0.05);②细胞克隆形成能力:shRNA转染组细胞克隆形成能力显著低于阴性对照组和未转染组(P < 0.05);③细胞凋亡:shRNA转染组细胞凋亡高于阴性对照组和未转染组,促凋亡蛋白Bax水平高于未转染组、对照组,抗凋亡蛋白Bcl-2表达低于阴性对照组和未转染组;④结果表明:干扰抑制TACC3基因表达,可抑制口腔鳞癌干细胞的增殖,促进其凋亡。

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程
ORCID: 0000-0003-0679-3091(李永生)

关键词: 干细胞, 肿瘤干细胞, 人口腔鳞癌, TACC3, 增殖, 凋亡

Abstract:

BACKGROUND: Studies have indicated that the abnormal expression of TACC3 is closely related to the occurrence and development of many kinds of tumors, and the expression of TACC3 is up-regulated in these tumors. Therefore, in vitro specific inhibition of TACC3 expression may become an important target for the treatment or intervention of tumor growth.
OBJECTIVE: To investigate the mechanism by which TACC3 gene expression regulates cell proliferation and apoptosis in oral squamous cell carcinoma.
METHODS: CD133+CD44+ oral squamous cell carcinoma cells were sorted from human oral squamous cell carcinoma cell line Cal-27 by immunomagnetic beads. In experimental group, the shRNA sequence of TACC3 was designed and synthesized, which was then trasnfected into CD133+CD44+ oral cancer stem cells by LipofectamineTM 2000. Empty vector-trasnfected (negative control) and untransfected cells were used as callsed. Forty-eight hours after the transfection, effects of TACC3 gene silencing on proliferation and apoptosis in vitro in CD133+CD44+ oral squamous cell carcinoma were detected by MTT, clone formation test, and TUNEL assay. Western blot assay was used to detect the effect of TACC3 gene silencing on Ki67, Bax and Bcl-2 protein expression in CD133+CD44+ oral squamous cell carcinoma.
RESULTS AND CONCLUSION: (1) Cell proliferation. The proliferation rate and expression level of Ki67 were significantly lower in the experimental group than the negative control and untransfected groups (P < 0.05). (2) Clone formation. The clone formation ability in the experimental group was significantly lower than that in the negative control and untransfected groups (P < 0.05). (3) Cell apoptosis. TACC3 gene silencing caused an obvious decrease in Bcl-2 protein expression and a significant increase in Bax protein expression. These findings further confirmed that specific interference of TACC3 gene expression could inhibit the proliferation of CD133+CD44+ cells and promote the apoptosis.

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

Key words: Neoplastic Stem Cells, Mouth Neoplasms, Cell Proliferation

中图分类号: