下调DNA结合分化抑制蛋白1基因对人结肠癌SW480细胞株体外转移和侵袭能力的影响研究

发布时间：2018-05-03 03:06

本文选题：结肠肿瘤 + 分化抑制蛋白　；参考：《中国全科医学》2017年08期

【摘要】：背景既往研究已经证实DNA结合分化抑制蛋白1(Id-1)表达升高在结直肠癌的发生、发展中发挥重要作用,但Id-1对结肠癌细胞侵袭行为影响及机制的研究较少。目的探讨下调Id-1基因对人结肠癌SW480细胞株增殖、转移、侵袭能力的影响及作用机制。方法 2016年1—6月,人结肠癌SW480细胞株分为3组:空白组,未进行转染;空载体组,用空载体进行转染;抑制组,采用Id-1特异小干扰RNA(siRNA)进行转染,采用反转录聚合酶链式反应(RT-PCR)和Western blotting法检测各组细胞Id-1和Ki-67基因及蛋白的表达情况,采用MTT法检测细胞增殖能力,采用细胞划痕实验、Transwell小室和Matrigel侵袭实验评价Id-1对细胞转移和侵袭能力的影响。结果 3组细胞Id-1、Ki-67 mRNA及其相应蛋白表达水平比较,差异均有统计学意义(P0.05);抑制组细胞Id-1、Ki-67 mRNA及其相应蛋白表达水平较空白组、空载体组降低(P0.05)。培养第1、2天,3组细胞增殖吸光度(OD值)比较,差异均无统计学意义(P0.05)。培养第3~7天,3组细胞增殖OD值比较,差异均有统计学意义(P0.05);其中抑制组细胞增殖OD值较空白组及空载体组降低(P0.05)。细胞划痕实验结果显示,抑制组细胞缓慢向中央迁移,缺损处修复较缓慢,空白组和空载体组细胞向划痕处的迁移速度明显加快,而空白组与空载体组间的迁移速度差异不明显。Transwell小室和Matrigel侵袭实验表明,3组迁移细胞数量和侵袭细胞数量比较,差异均有统计学意义(P0.05);抑制组迁移细胞数量和侵袭细胞数量较空白组、空载体组减少(P0.05)。结论下调Id-1基因能抑制人结肠癌SW480细胞株的增殖、转移和侵袭能力,其机制可能与抑制Ki-67的表达有关。
[Abstract]:Background research has confirmed that the increase of DNA binding inhibitor protein 1 (Id-1) expression plays an important role in the development of colorectal cancer. However, there are few studies on the impact of Id-1 on the invasion of colon cancer cells and its mechanism. The purpose of this study is to investigate the effect of down regulation of Id-1 gene on the proliferation, metastasis and invasion of human colon cancer cell line. Methods from 1 to June 2016, human colon cancer SW480 cell lines were divided into 3 groups: blank group, untransfected, empty carrier group, transfected with empty carrier, inhibition group, Id-1 specific small interference RNA (siRNA) transfection, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting method to detect Id-1 and Ki-67 genes and eggs of each group. In white expression, MTT assay was used to detect cell proliferation, cell scratch test, Transwell chamber and Matrigel invasion test to evaluate the effect of Id-1 on cell metastasis and invasion. Results 3 groups of cells, Id-1, Ki-67 mRNA and corresponding protein expression levels were compared, the difference was statistically significant (P0.05), Id-1, Ki-67 in the inhibition group, Ki-67. The expression level of mRNA and its corresponding protein was lower than that in the blank group (P0.05). There was no significant difference between the 3 groups of cell proliferation absorbency (P0.05). On day 3~7, the proliferation of cells in the 3 groups was statistically significant (P0.05), and the proliferation of cell proliferation in the inhibition group was more than that in the blank group and the empty body. The group decreased (P0.05). The results of cell scratch test showed that the cells migrated slowly to the center and the repair of the defect was slow. The migration speed of the cells in the blank group and the empty carrier group was quicker than that of the blank group and the empty carrier group. The difference of the migration velocity between the blank group and the empty carrier group was not obvious in the.Transwell chamber and the Matrigel invasion experiment, which showed that the 3 groups migrated fine. The number of cells and the number of invasive cells were statistically significant (P0.05); the number of cells and the number of invasive cells in the inhibition group were less than that in the blank group, and the number of the unloaded body decreased (P0.05). Conclusion the down-regulation of Id-1 gene could inhibit the proliferation, metastasis and invasion of human colon cancer SW480 cell lines, and the mechanism may be related to the inhibition of the expression of Ki-67.

【作者单位】：河北北方学院附属第一医院血管腺体外科;河北北方学院附属第一医院超声科;河北北方学院附属第一医院胃肠外科;河北北方学院附属第一医院病理科;
【基金】：河北省科技支撑计划资助项目(152777237) 河北省卫计委医学科学研究重点课题计划(20150058) 河北省张家口市科技局指令性计划(1311055D)
【分类号】：R735.35

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