口腔疾病防治 ›› 2020, Vol. 28 ›› Issue (9): 562-568.DOI: 10.12016/j.issn.2096-1456.2020.09.003

• 基础研究 • 上一篇    下一篇

TGF-β1对口腔癌相关成纤维细胞在二维和三维共培养条件下迁移的影响

杨津1(),吴飞飞1(),高庆红2,李小玉3,MANABUKato4,程然5(),周红梅1()   

  1. 1.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院口腔黏膜病科,四川 成都(610041)
    2.四川大学华西口腔医院口腔颌面外科,四川 成都(610041)
    3.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院,四川 成都(610041)
    4.三重大学医院肾内科,日本 三重县(514-8507)
    5.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院预防科,四川 成都(610041)
  • 收稿日期:2019-12-09 修回日期:2020-01-22 出版日期:2020-09-20 发布日期:2020-08-24
  • 通讯作者: 程然,周红梅
  • 作者简介:杨津,医师,博士,Email:2012181643016@stu.scu.edu.cn|吴飞飞,医师,硕士,Email:wufei1122_qi@163.com
    杨津和吴飞飞为共同第一作者
  • 基金资助:
    国家自然科学基金资助项目(81772898)

Effects of TGF-β1 on the migration of oral cancer-associated fibroblasts in two and three dimensional co-culture models

YANG Jin1(),WU Feifei1(),GAO Qinghong2,LI Xiaoyu3,MANABU Kato4,CHENG Ran5(),ZHOU Hongmei1()   

  1. 1. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Oral Medicine, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
    2. Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
    3. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
    4. Urology, Mie University Hospital, Mie 514-8507, Japan
    5. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Department of Preventive Dentistry, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2019-12-09 Revised:2020-01-22 Online:2020-09-20 Published:2020-08-24
  • Contact: Ran CHENG,Hongmei ZHOU

摘要:

目的 通过二维和三维条件下的细胞共培养模型观察转化生长因子-β1(transforming growth factor-β1,TGF-β1)对口腔癌相关成纤维细胞(carcinoma associated fibroblasts,CAFs)迁移的影响。方法 二维培养条件下,以培养基中加入10 ng/mL TGF-β1刺激的CAFs为实验组,以未经处理的CAFs细胞为对照组,通过划痕实验、Transwell实验观察CAFs的迁移现象;通过逆转录病毒转染,培养绿色荧光蛋白(green fluorescent protein,GFP)阳性的CAFs细胞,建立人舌鳞癌细胞SCC25、GFP(+) CAFs和CAFs三维细胞共培养模型,以培养基中加入10 ng/mL TGF-β1刺激下培养的三维模型为实验组,以未加的TGF-β1的三维模型为对照组,观察在三维模型的细胞迁移现象。结果 在二维培养条件下验证了10 ng/mL TGF-β1可促进口腔CAFs的迁移;成功建立SCC25、GFP(+) CAFs和CAFs三维细胞共培养模型,观察到CAFs和口腔癌细胞的迁移现象,但10 ng/mL的TGF-β1对上述细胞的迁移在三维条件下无明显促进作用。结论 体外TGF-β1对口腔CAFs迁移的影响与二维和三维不同培养模式有关。

关键词: 癌相关成纤维细胞, 迁移, 三维细胞共培养模型, 转化生长因子-β1, 口腔癌

Abstract:

Objective To observe the effect of transforming growth factor-β1 (TGF-β1) on the migration of oral carcinoma associated fibroblasts (CAFs) with two-dimensional culture model and three-dimensional model.Methods Under two-dimensional culture conditions, CAFs stimulated by TGF-β1 with the addition of 10 ng/mL medium were used as the experimental group, and untreated CAFs were used as the control group. The migration of CAFs with the stimulation of TGF-β1 was measured by cell scratch assay and transwell assay. CAFs positive for green fluorescent protein (GFP) were cultured by retrovirus transfection. Human tongue squamous cell carcinoma cells SCC25, GFP(+) CAFs and CAFs with three-dimensional cell co-culture models were established. The three-dimensional model cultured under the stimulation of TGF-β1 with 10 ng/mL medium was used as the experimental group, and the three-dimensional model without TGF-β1 was used as the control group. The migration of CAFs with the stimulation of TGF-β1 was also measured by the three-dimensional models.Results It was verified that 10 ng/mL TGF-β1 promoted the migration of CAFs in the two-dimensional culture model. The three-dimensional co-culture models of SCC25, GFP(+) CAFs and CAFs were successfully established. The migration of SCC25 and CAFs was detected in the three-dimensional model. However, 10 ng/mL TGF-β1 had little effect on their migration.Conclusion The effect of TGF-β1 in vitro on the migration of oral CAFs was associated with different culture models in two and three dimensions.

Key words: carcinoma-associated fibroblasts, migration, three-dimensional cell culture model, transforming growth factor-β1, oral carcinoma

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