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观察空间诱变后恶性黑色素瘤B16细胞株的细胞间通讯功能及细胞间黏附能力的变化,并探讨可能的机理。应用细胞低温长期生存系统,将B16细胞株搭载于“第20颗返回式卫星”,返地后单克隆化,从得到的110株单克隆空间诱变B16细胞株中随机选取6株,编号为39#～44#,常规培养6代后和对照细胞株同时做MTT和FCM,检测细胞增殖和周期分布;用划痕标记染料示踪技术检测细胞间通讯功能;将有变异的细胞接种C57BL/6J小鼠,免疫组化观测CD31蛋白的表达量。初步筛选出两株(39#、44#)变化比较明显的细胞株,44#细胞增殖能力降低、G1期分布增加、细胞间通讯功能增强、CD31蛋白表达增强;39#细胞增殖能力升高、G1期分布减少、细胞间通讯功能减弱、CD31蛋白表达减少。空间环境的作用使体外培养肿瘤细胞产生多向变异,有可能筛选出有意义的细胞株做进一步的研究。
To observe the changes of intercellular communication and intercellular adhesion in malignant melanoma B16 cell line after mutagenesis, and to explore the possible mechanism. The cell cryogenic long-term survival system was used. The B16 cell line was mounted on the “20th returning satellite” and returned to the ground for single cloning. Six of 110 strains of monoclonal space-mutated B16 cells were randomly selected and identified as 39 # ~ 44 #. After 6 passages of conventional culture, MTT and FCM were performed simultaneously with the control cell lines to detect cell proliferation and cell cycle distribution. Scratch-mark dye-tracing technique was used to detect the cell-to-cell communication function. Inoculated cells with mutation C57BL / 6J mice, the expression of CD31 protein was observed by immunohistochemistry. The results showed that the proliferation of 44 # cells was decreased, the distribution of G1 phase was increased, the intercellular communication function was enhanced and the expression of CD31 protein was enhanced. The proliferation of 39 # G1 phase distribution decreased, intercellular communication function weakened, CD31 protein expression decreased. The role of space environment in vitro tumor cells to produce multi-directional variation, it is possible to screen out meaningful cell lines for further study.