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目的应用基因芯片技术对小鼠组织细胞肉瘤L-Ⅱ高侵袭转移瘤株和母瘤株的基因表达谱进行对比分析并筛选出差异表达基因,以期从基因水平上探讨肿瘤的转移机制。方法 L-Ⅱ瘤细胞接种小鼠胁部皮下,取其肺转移灶制成细胞悬液接种另一小鼠体内,再取其肺转移灶传代,如此数代后,建立L-Ⅱ肺高转移模型,筛选出L-Ⅱ高侵袭转移瘤株。用基因芯片检测筛选前后的L-Ⅱ瘤株表达有差异的基因,并用RT-PCR对部分差异表达基因进行分析鉴定。结果用体内转移灶连续传代法筛选小鼠组织细胞肉瘤L-Ⅱ至第9代,筛选出L-Ⅱ高侵袭转移瘤株。用肿瘤转移基因芯片杂交筛选出传代前后L-Ⅱ瘤细胞的差异表达基因共21个,其中上调基因17个,下调基因4个。对其中有代表意义的基因,如Kras-2、IGF-1、MMP-10、Brms-1进行RT-PCR验证,结果证实所选基因在筛选前后瘤细胞中的表达差异均有统计学意义,与基因芯片的表达情况一致。结论小鼠组织细胞肉瘤L-Ⅱ的侵袭转移是一个涉及Kras-2、IGF-1、MMP-10、Brms-1等重要基因和其他相关基因等多个基因共同参与的复杂过程。
OBJECTIVE: To compare the gene expression profile of L-Ⅱ highly-invasive tumor cell line and tumorous tissue of mice with histiocytic sarcoma by gene chip technique and screen the differentially expressed genes in order to explore the mechanism of tumor metastasis at gene level. Methods L-Ⅱ tumor cells were inoculated subcutaneously in the flank of the mice and their lung metastases were made into a cell suspension to inoculate another mouse and passaged from their lung metastases. After several generations, L-Ⅱ lung hyperplasia Model, screened L-Ⅱ highly invasive metastatic tumor strains. Genes were used to detect the differentially expressed genes of L-II tumor before and after screening, and some differentially expressed genes were identified by RT-PCR. Results The passage cell line L-Ⅱ of mice was screened by continuous passage in vivo to screen the L-Ⅱ highly invasive metastatic tumor line. Twenty-one differentially expressed genes of L-II tumor cells were screened by tumor metastasis gene chip hybridization, including 17 up-regulated genes and 4 down-regulated genes. RT-PCR validation of the representative genes such as Kras-2, IGF-1, MMP-10 and Brms-1 showed that there was significant difference in the expression of the selected genes in the tumor cells before and after screening, Consistent with the gene chip expression. Conclusions The invasion and metastasis of murine myeloid sarcoma L-Ⅱ is a complex process involving multiple genes such as Kras-2, IGF-1, MMP-10, Brms-1 and other related genes.