[人喉癌Hep-2细胞系中CD133+癌干细胞生物学特性的研究]

Wei Xu-Dong, He Jian, Gao Jiang-Xia, Wang Jing-Yu, Ma Bin-Juan, Chen Jing

Department of Otolaryngology Head and Neck Surgery, Gansu Provincial Hospital, Lanzhou, China.

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2013 Jul;48(7):578-83.

OBJECTIVE

To investigate the biology characters of CD133+ cancer stem cells from Hep-2 cell line.

METHODS

The Flow cytometry was applied to purify CD133+ cells from Hep-2 cell line. The sorted CD133+ cells were cultured in RPMI1640. The ability of migration, invasion and clonality of CD133+ cells were performed to characterize the properties of the cells. CD133+ cells and CD133- control cells were treated with paclitaxel and exposed respectively to X-rays emitted by linear accelerator with a dose of 10 Gy. The surviving rates and growth inhibition ratio of cells in two groups were detected with MTT assay to observe the resistance to irradiation and chemotherapy in CD133+ cells.

RESULTS

The percentage of CD133+ cells in the unsorted and the sorted cells were 3.1% +/- 0.2% and 90.2% +/- 5.5%, respectively. CD133+ cells showed the higher proliferation and colony ability than CD133- cells. The numbers of CD133+ and CD133- cells that passed the membrane of Transwell chamber were 526 +/- 39 and 220 +/- 20 respectively (t = 22.08, P < 0.001). The colony forming units of three passages were 30.0% +/- 4.7%, 32.2% + 3.6%, 32.7% + 3.4% in CD133 cells and 15.2% +/- 2.2%, 12.0% + 2.5%, 13.8% +/- 3.3% in CD133- cells. There were statistic difference between two groups (t = 8.99, t = 14.66, t = 12.69, P < 0.01). At 24, 48, and 72 hours of treatment with paclitaxel, the cell surviving rates of CD133+ cells were 90.1% +/- 5.9%, 85.1% +/- 7.1% and 70.3% +/- 6.4% and lower than those of CD133- cells, respectively (t = 5.24, t = 8.18, t = 8.14, P < 0.01) . After radiotherapy, growth inhibition ratio of CD133+ and CD133- cells were 30.0% +/- 7.1% and 55.0% +/- 6.3% (t = 8.30, P < 0.01).

CONCLUSIONS

CD133+ cells exist in a small proportion in Hep-2 cell line and they show the properties of cancer stem cells, with the resistance to irradiation and chemotherapy.

目的

研究人喉癌细胞系Hep-2中CD133+肿瘤干细胞的生物学特性。

方法

应用流式细胞术从Hep-2细胞系中分离纯化CD133+细胞，将分选后的CD133+细胞接种于RPMI1640培养基中培养，通过检测其迁移、侵袭及克隆形成能力来鉴定细胞特性。分别用紫杉醇处理CD133+细胞和CD133-对照细胞，并给予直线加速器10 Gy X射线照射，采用MTT法检测两组细胞的存活率和生长抑制率，观察CD133+细胞对放疗和化疗的抵抗性。

结果

未分选细胞和分选后细胞中CD133+细胞所占比例分别为3.1%±0.2%和90.2%±5.5%。CD133+细胞较CD133-细胞具有更强的增殖和克隆能力。穿过Transwell小室膜的CD133+细胞和CD133-细胞数分别为526±39和220±20(t = 22.08，P < 0.001)。CD133+细胞传代3次后的克隆形成率分别为30.0%±4.7%、32.2%±3.6%、32.7%±3.4%，CD133-细胞分别为15.2%±2.2%、12.0%±2.5%、13.8%±3.3%，两组比较差异有统计学意义(t = 8.99、t = 14.66、t = 12.69，P < 0.01)。紫杉醇作用24、48和72 h后，CD133+细胞的存活率分别为90.1%±5.9%、85.1%±7.1%和70.3%±6.4%，均低于CD133-细胞(t = 5.24、t = 8.18、t = 8.14，P < 0.01)。放疗后，CD133+细胞和CD133-细胞的生长抑制率分别为30.0%±7.1%和55.0%±6.3%(t = 8.30，P < 0.01)。