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目的:比较CD133+和CD133-胶质母细胞瘤体内再移植的细胞性征。方法:剥离26只CD133+和2只CD133-胶质母细胞瘤细胞致瘤裸鼠皮下肿瘤组织,分成4份,第1部分肿瘤块制作成冰冻切片,原位凋亡试剂盒检测细胞凋亡率;从第2部分组织块中提取细胞总蛋白,蛋白质印迹法检测转录因子Oct4和Sox2、细胞核增殖抗原(PCNA)、表皮生长因子受体(EGFR)、血管生成因子2(Ang-2)、基质金属蛋白酶2(MMP-2)和MMP-9的表达水平;第3部分组织块剪碎消化成单细胞悬液,流式细胞术检测CD133+肿瘤细胞百分率;取第4部分肿瘤组织进行去胸腺裸鼠皮下再移植,检测成瘤率。结果:CD133+胶质母细胞瘤细胞所致肿瘤组织细胞凋亡率(4.37±0.74)%,低于CD133-胶质母细胞瘤细胞所致肿瘤组织细胞凋亡率(11.14±2.15)%,差异有统计学意义,P<0.05;CD133+胶质母细胞瘤组织Oct4、Sox2、PCNA、EGFR、Ang2、MMP-2和MMP-9蛋白表达水平相对较高;CD133+和CD133-胶质母细胞瘤细胞所致肿瘤中CD133+细胞百分率分别为(2.47±0.67)%和(0.44±0.14)%,两者致裸鼠皮下成瘤率分别为10/10和4/10。结论:CD133+胶质母细胞瘤细胞所致肿瘤较CD133-胶质母细胞瘤细胞所致肿瘤具有更高的恶性表型,两者体内再移植后成瘤率提高。
OBJECTIVE: To compare the cellular signs of in vivo re-transplantation of CD133 + and CD133-glioblastoma. METHODS: Twenty-six CD133 + and two CD133-glioblastoma tumor-bearing mice were dissected and subcutaneously divided into four groups. The first part of tumor mass was made into frozen sections, and the apoptosis rate was detected by in situ apoptosis kit ; Total cellular protein was extracted from the second part of the tissue mass, and the transcription factors Oct4 and Sox2, PCNA, EGFR, Ang-2 were detected by Western blotting, The expression of MMP-2 and MMP-9 were detected by immunohistochemistry. The third part of the tissue pieces were cut and digested into single cell suspension, and the percentage of CD133 + tumor cells was determined by flow cytometry. Rats were subcutaneously transplanted to detect tumor formation rate. Results: The apoptosis rate of tumor cells induced by CD133 + glioblastoma cells was (4.37 ± 0.74)%, which was lower than that of CD133-glioblastoma cells (11.14 ± 2.15)%, the differences (P <0.05). The expressions of Oct4, Sox2, PCNA, EGFR, Ang2, MMP-2 and MMP-9 in CD133 + glioblastoma tissues were higher than those in CD133 + (2.47 ± 0.67)% and (0.44 ± 0.14)%, respectively. The subcutaneous tumorigenic rates of the two groups were 10/10 and 4/10, respectively. Conclusion: The tumors induced by CD133 + glioblastoma cells have a higher malignant phenotype than that of CD133-glioblastoma cells.