结直肠癌中tcf21调控肿瘤转移抑制基因kiss-1表达的分析-analysis of tcf 21 regulating the expression of kiss - 1 in colorectal cancer.docxVIP

达水平均与患者的淋巴结转移、远处器官及临床TNM分期有关（P＜0.05）；但与患者的性别、年龄、分化程度、病理类型、侵润深度、脉管内癌栓、瘤体大小、 坏死等无关（P＞0.05）；二者的表达存在着明显的正相关性（r=0.450，P＜0.05）。 2、结直肠癌细胞株HT-29中TCF21与Kiss-1基因mRNA的表达水平均相对高于 SW480，Lovo细胞株。3、转染了siRNA的HT-29细胞3组，si1组中TCF21 mRNA与蛋白的表达水平均明 显降低，与对照组间差异有统计学意义（P＜0.05），TCF21基因被成功沉默。 4、si1组中Kiss-1 mRNA和蛋白的表达水平亦明显降低，与对照组间差异具有统 计学意义（P＜0.05），Kiss-1基因的表达被下调。5、siRNA成功沉默HT-29细胞后，CCK-8法显示细胞增殖能力明显增强 (P＜0.01)；Transwell小室实验显示，细胞侵袭及迁移能力亦明显增强(P＜0.01)。 结论：1、TCF21 与 Kiss-1 基因的表达存在着明显的正相关性，二者的表达缺失与结直 肠癌的进展过程密切相关，并可能是结直肠癌远处转移风险增高的促进因素。 2、沉默结直肠癌 HT-29 细胞株 TCF21 基因的表达，可引起 Kiss-1 基因 mRNA 和蛋白表达水平的共同下降，并导致细胞增殖、侵袭及迁移能力的增强。 3、结直肠癌病变进展过程中 Kiss-1 基因可能是作为下游功能基因，接受 TCF21 基因的转录调控而发挥其肿瘤转移抑制效应。关键词：结直肠癌，TCF21，Kiss-1，相关性，调控Research of TCF21 regulate tumor metastasis suppressor gene Kiss-1 expression in colorectal cancerSpecialtySurgery PostgraduateChen ChangjiangTutor Prof.Dai QibaoChen ShaoqinABSTRACTObjective:Analyse the influence affected by tumor metastasis suppressor gene TCF21 and Kiss-1 on Clinicopathological features of colorectal cancer; discusses the correlation between their expression.Construct an expression plasmid of siRNA against TCF21 gene in human colorectal cancer cell line HT- 29; to investigate its impact to the expression of Kiss-1 gene, and observe the changes to the cell biological behavior of proliferation, invasion and migration after inhibited.Methods:The expression of TCF21 and Kiss-1 gene were examined by IHC two step method in 71 specimens of colorectal primary cancer tissues together with normal tissues adjacent to cancer.Adopt qRT-PCR method to choose the relatively high expression of TCF21 and Kiss-1 mRNA from colorectal cancer cell lines among Lovo, SW480, HT-29.Design three specific siRNA molecule fragments for TCF21, and transfected into HT-29 cells by lipofectamineTM2000; adopt qRT-PCR method to test the expression level of TCF21 mRNA after 48h, pick out the highest inhibition effective fragment; adopt Western-blot method to test the expression level of TCF21 protein after 72h, i