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目的筛选经太空环境诱变后B16细胞致瘤性改变的细胞株,观察其基因表达的改变,寻找与黑色素瘤发生及转移密切相关的基因。方法将小鼠黑色素瘤B16细胞搭载于第20颗返回式卫星,返地后克隆化,随机选择5株克隆细胞株(1#、5#、6#、7#和8#),接种C57BL/6J小鼠,分别观察细胞致瘤性、荷瘤小鼠生存期及瘤体病理变化,用基因表达谱分析其中2株初筛致瘤性改变细胞株(1#和8#)的基因表达情况。结果与对照组比较,1#细胞株接种小鼠后,肿瘤潜伏期延长(P<0·05),1#和8#细胞株移植瘤瘤体重量减小(P<0·01),且荷瘤小鼠生存期延长(P<0·01);病理诊断结果显示:1#和8#细胞移植瘤内血管不及对照组丰富,坏死区较少,瘤体内浸润淋巴细胞较对照组有不同程度的增多,瘤旁组织内有较多淋巴细胞;基因芯片分析结果表明:与对照B16细胞相比,2株诱变细胞(1#和8#)分别有145和125个基因表达水平发生改变(P<0·05),其中9个基因为共同变化基因,前列腺素D2合成酶基因在2株细胞中的表达均明显增高,与对照相比,差别在5倍以上。结论1#和8#细胞株致瘤性减弱,且2株细胞的移植瘤肿瘤新生血管及癌旁浸润减少,癌旁组织中有较多淋巴细胞,推测可能与前列腺素D2合成酶等多种肿瘤相关基因表达的改变有关。
Objective To screen the cell lines with tumorigenicity induced by space environment mutagenesis and to observe the gene expression changes and to find out the genes closely related to the occurrence and metastasis of melanoma. Methods Mouse melanoma B16 cells were mounted on the 20th retrospective satellite and returned to ground. Five cloned cell lines (1 #, 5 #, 6 #, 7 # and 8 #) were randomly selected and inoculated with C57BL / 6J mice were used to observe the tumorigenicity of tumor-bearing mice, the survival of tumor-bearing mice and the pathological changes of tumor. The gene expression profiles of 2 primary screening tumorigenicity cell lines (1 # and 8 #) were analyzed by gene expression profiling . Results Compared with the control group, the tumor latency of 1 # cell line was prolonged (P <0.05), while the tumor weight of 1 # and 8 # cell lines reduced (P <0.01) (P <0.01). The results of pathological diagnosis showed that the number of blood vessels in 1 # and 8 # cells was not as large as that in the control group, the number of necrotic areas was less, and the number of infiltrating lymphocytes in the tumor tissue was different from that of the control group , And there were more lymphocytes in the tumor-adjacent tissues. Gene chip analysis showed that 145 and 125 gene expression levels were changed in the two mutated cells (# 1 and # 8) compared with the control B16 cells ( P <0.05). Nine genes were common and the expression of prostaglandin D2 synthase gene was significantly increased in both cell lines, which was more than 5 times higher than the control. Conclusions The tumorigenicity of 1 # and 8 # cell lines is weakened, and the transplanted neoplasms and para-cancerous lesions of 2 cells are reduced, and there are more lymphocytes in the paracancerous tissues, which may be related to the prostaglandin D2 synthase Tumor-related gene expression changes.