Notch信号途径关键转录因子RBP-JK基因敲除结肠癌细胞株CT-26的体内外增殖能力观察  

Proliferative ability in vivo and vitro of colon cancer cell line CT-26 with Notch signaling pathway key transcription factor RBP-JK knockout

作　　者：王淑红[1] 王耀春 WANG Shuhong;WANG Yaochun(The First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China)

机构地区：[1]西安交通大学第一附属医院

出　　处：《山东医药》2019年第33期19-22,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81672810);西安交通大学第一附属医院院基金项目(2019ZYTS-19)

摘　　要：目的探讨Notch信号途径关键转录因子重组信号蛋白-JK(RBP-JK)基因敲除的结肠癌细胞株CT-26的体外增殖能力及体内成瘤能力,以探讨Notch信号途径在结肠癌发病机制中的作用。方法通过敲除RBP-JK靶序列的慢病毒干扰抑制小鼠结肠癌细胞株CT-26中Notch信号途径关键性转录因子RBP-JK的表达制作RBP-JK基因敲除CT-26细胞株模型,鉴定模型制作成功后,采用BrdU掺入实验,根据相同时间内掺入BrdU的细胞比例大小,观察RBP-JK基因敲除CT-26细胞株及对照CT-26细胞株的体外增殖能力;通过体内成瘤实验、根据肿瘤生长体积大小观察RBP-JK基因敲除CT-26细胞株及对照CT-26细胞株在小鼠体内的成瘤能力。结果RBP-JK基因敲除CT-26细胞株、对照CT-26细胞株在培养1 h时掺入BrdU的细胞比例分别为15.21%±0.242%、10.78%±0.251%,二者比较,t=31.10,P<0.01;在培养3 h时掺入BrdU的细胞比例分别为25.46%±0.316%、13.26%±0.284%,二者比较,t=70.26,P<0.01。RBP-JK基因敲除CT-26细胞株接种后第24、27、30天,小鼠直肠原位肿瘤体积分别为(0.351±0.014)、(0.422±0.011)、(0.737±0.021)mm 3,对照CT-26细胞株接种后第24、27、30天,小鼠直肠原位肿瘤体积分别为(0.513±0.013)、(1.161±0.114)、(1.848±0.016)mm 3,二者同时间比较,t分别为25.61、15.84、101.10,P均<0.01。结论RBP-JK基因可阻断结肠癌细胞株CT-26的Notch信号途径,敲除RBP-JK基因敲除CT-26细胞株体外增殖能力及体内成瘤能力均降低。Objective To investigate the proliferation in vitro and tumorigenesis in vivo of colorectal cancer cell line CT-26 with Notch signaling pathway key transcription factor recombinant signal protein-JK(RBP-JK)knockout,and to explore the role of Notch signaling pathway in the pathogenesis of colon cancer.Methods RBP-JK knockout CT-26 cell model was established.After the model was established successfully,BrdU incorporation experiment was used to assess the proliferation of RBP-JK gene knockout CT-26 cell line and control CT-26 cell line by the BrdU incorporation cell percentage within the same time.The tumor-forming ability of RBP-JK gene knockout CT-26 cell line and control CT-26 cell line in mice was evaluated by observing the tumor growth volume in vivo and in vivo.Results The BrdU incorporation cell percentages of RBP-JK gene knockout CT-26 cell line and control CT-26 cell line within 1 h were 15.21%±0.242%and 10.78%±0.251%,respectively(t=31.10,P<0.01),and were 25.46%±0.316%and 13.26%±0.284%,respectively,within 3 h(t=70.26,P<0.01).On the 24th,27th and 30th days after the inoculation of RBP-JK gene knockout CT-26 cell line in mice rectum in situ,the tumor volume was(0.351±0.014),(0.422±0.011),and(0.736±0.021)mm 3,respectively;the tumor volume of control CT-26 cell line in mice was(0.513±0.012),(1.161±0.113),and(1.847±0.016)mm 3,respectively,and there was significant difference(t=25.61,15.84,and 101.1,respectively;all P<0.01).Conclusion Knocking out RBP-JK gene can block Notch signaling pathway of CT-26 cell line,and both the proliferation in vitro and tumorigenesis in vivo of RBP-JK gene knockout colorectal cancer cells CT-26 decrease.

关 键 词：NOTCH信号途径 重组信号蛋白-JK 结肠癌细胞 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]