小鼠骨髓间充质干细胞对人舌鳞状细胞癌Cal27细胞的归巢作用及其对Cal27细胞增殖和迁移的促进作用

doi:10.13481/j.1671-587x.20180501

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (05): 891-896.doi: 10.13481/j.1671-587x.20180501

• 基础研究 • 下一篇

小鼠骨髓间充质干细胞对人舌鳞状细胞癌Cal27细胞的归巢作用及其对Cal27细胞增殖和迁移的促进作用

孟琳^1,2, 王璐^1,2, 布文奂^1,2, 丁鑫鑫³, 高华丽⁴, 王梓霖^1,2, 刘玉兰^1,2, 李琛⁵, 孙宏晨^1,2

1. 吉林省牙发育及颌骨重塑与再生重点实验室, 吉林长春 130021;
2. 吉林大学口腔医院病理科, 吉林长春 130021;
3. 吉林大学口腔医院种植科, 吉林长春 130021;
4. 吉林大学口腔医院儿童口腔科, 吉林长春 130021;
5. 吉林大学口腔医院口腔黏膜病科, 吉林长春 130021

收稿日期:2017-12-28 出版日期:2018-09-28 发布日期:2018-11-20
通讯作者: 孙宏晨,教授,博士研究生导师(Tel:0431-88796010,E-mail:hcsun@mail.jlu.edu.cn) E-mail:hcsun@mail.jlu.edu.cn
作者简介:孟琳(1993-),女,陕西省西安市人,在读医学硕士,主要从事纳米材料载药/载基因抗肿瘤方面的研究。
基金资助:
国家自然科学基金国际合作与交流项目资助课题（8132010801）；吉林省科技厅省级产业创新专项资金项目资助课题（2016C044-3）；吉林省科技厅科技发展计划项目资助课题（20160101347JC）；吉林省科技厅自然科学基金项目资助课题（20170101093JC）

Homing effect of mouse bone marrow-derived mesenchymal stem cells on human tongue squamous cell carcinoma Cal27 cells and its promotion on proliferation and migration of Cal27 cells

MENG Lin^1,2, WANG Lu^1,2, BU Wenhuan^1,2, DING Xinxin³, GAO Huali⁴, WANG Zilin^1,2, LIU Yulan^1,2, LI Chen⁵, SUN Hongchen^1,2

1. Key Laboratory of Tooth Development and Bone Remodeling and Regeneration, Jilin Province, Changchun 130021, China;
2. Department of Oral Pathology, Stomatology Hospital, Jilin University, Changchun 130021, China;
3. Department of Dental Implantology, Stomatology Hospital, Jilin University, Changchun 130021, China;
4. Department of Pediatric Dentistry, Stomatology Hospital, Jilin University, Changchun 130021, China;
5. Department of Oral Mucosal Disease, Stomatology Hospital, Jilin University, Changchun 130021, China

Received:2017-12-28 Online:2018-09-28 Published:2018-11-20

摘要/Abstract

摘要： 目的：探讨小鼠骨髓间充质干细胞（mBMSCs）对舌鳞状细胞癌Cal27细胞的归巢作用，并观察mBMSCs影响下舌鳞状细胞癌Cal27细胞的生物学行为。方法：以全骨髓法分离提纯mBMSCs，采用流式细胞术分析细胞表型进行鉴定。将mBMSCs分为空白对照组、Hacat细胞对照组和Cal27细胞实验组，将正常培养基、Hacat细胞条件培养基和Cal27细胞条件培养基分别与mBMSCs共培养后，通过细胞迁移实验检测mBMSCs迁移的细胞数。再将Cal27细胞分为单纯Cal27细胞对照组和与mBMSCs共培养Cal27细胞实验组（共培养组），共培养组使用Transwell法共培养Cal27细胞和mBMSCs，采用细胞增殖实验和克隆形成实验检测共培养后Cal27细胞的增殖能力；细胞迁移实验检测共培养后Cal27细胞的迁移能力。结果：倒置显微镜观察，mBMSCs多数细胞呈短梭形，细胞质丰富，细胞核呈椭圆形或肾形，细胞突长短不一，细胞排列整齐。流式细胞术检测，mBMSCs表面标志物CD44、CD29和Sca-1呈阳性，CD45和CD11b呈阴性。与空白对照组和Hacat细胞对照组比较，Cal27组细胞实验归巢至Cal27细胞的mBMSCs数量明显增加（P<0.01）。与单纯Cal27细胞对照组比较，共培养组Cal27细胞从共培养第3天起生长速度明显高于对照组（P<0.01），克隆形成数增加。细胞迁移实验，共培养组Cal27细胞穿膜细胞数较单纯Cal27细胞对照组明显增加（P<0.01）。结论：mBMSCs可归巢至舌鳞状细胞癌，促进舌鳞癌Cal27细胞的增殖和迁移，从而促进舌鳞癌的发展，提示mBMSCs可作为抗肿瘤治疗靶点。

关键词: 骨髓间充质干细胞, 口腔鳞状细胞癌, 共培养, 归巢

Abstract: Objective:To explore the homing effect of mouse bone marrow-derived mesenchymal stem cells (mBMSCs) on the tongue squamous cell carcinoma Cal27 cells, and to observe the biological behaviors of tongue squamous cell carcinoma Cal27 cells affected by mBMSCs. Methods:The mBMSCs were isolated and purified by whole bone marrow methods. The cell phenotype was analyzed by flow cytometry to identify the mBMSCs. The mBMSCs were divided into blank control group, Hacat cell control group and Cal27 cell experiment Cal27 cell group. After co-cultured with normal culture medium, Hacat conditioned medium and Cal27 conditioned medium,the number of the migrated Cal27 cells was detected by cell migration assay. The Cal27 cells were divided into single Cal27 cell control group and co-cultured Cal27 and mBMSCs experiment group(co-cultured group). The Cal27 cells in co-cultured group were co-cultured with mBMSCs using Transwell method. Cell proliferation assay and colony formation assay were used to detect the proliferation ability of the co-cultured Cal27cells. Cell migration assay was performed for the detection of the migration ability of the Cal27 cells after co-culture. Results:The inverted microscope results showed that most mBMSCs were short shuttle-like, with abundant cytoplasm, oval or nephron nucleus and irregular cell lengths; the cells arranged well. The results of flow cytometry showed that CD44, CD29, and Sca-1 were positive, and CD45, CD11b were negative in the mBMSCs. Compared with blank control group and Hacat cell control group, the number of mBMSCs homing to Cal27 cells in Cal27 cell group was significantly increased (P<0.01). Compared with single Cal27 cell control group, the growth rates of Cal27 cells in co-cultured group were significantly increased from the 3rd day after co-culture(P<0.01), and the number of clones was increased. The cell migration test results showed that the number of transmembrane cells in co-cultured group was significantly increased compared with single Cal27 cell control group (P<0.01). Conclusion:mBMSCs can home to the tongue squamous cell carcinoma Cal27 cells, which promotes the proliferation and migration of tongue squamous cell carcinoma Cal27 cells and promotes the development of tongue squamous cell carcinoma. So mBMSCs can be used as an anti-tumor target.

Key words: bone marrow-derived mesenchymal stem cells, oral squamous cell carcinoma, co-culture, homing

中图分类号:

R73-37

孟琳, 王璐, 布文奂, 丁鑫鑫, 高华丽, 王梓霖, 刘玉兰, 李琛, 孙宏晨. 小鼠骨髓间充质干细胞对人舌鳞状细胞癌Cal27细胞的归巢作用及其对Cal27细胞增殖和迁移的促进作用[J]. 吉林大学学报(医学版), 2018, 44(05): 891-896.

MENG Lin, WANG Lu, BU Wenhuan, DING Xinxin, GAO Huali, WANG Zilin, LIU Yulan, LI Chen, SUN Hongchen. Homing effect of mouse bone marrow-derived mesenchymal stem cells on human tongue squamous cell carcinoma Cal27 cells and its promotion on proliferation and migration of Cal27 cells[J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 891-896.

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小鼠骨髓间充质干细胞对人舌鳞状细胞癌Cal27细胞的归巢作用及其对Cal27细胞增殖和迁移的促进作用

Homing effect of mouse bone marrow-derived mesenchymal stem cells on human tongue squamous cell carcinoma Cal27 cells and its promotion on proliferation and migration of Cal27 cells

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