端粒酶抑制剂1对裸鼠结直肠癌转移的影响

目的建立人结直肠癌裸鼠转移瘤模型，探讨端粒酶抑制剂1（PinX1）对结直肠癌转移的影响。方法分别通过PinX1干扰慢病毒、阴性对照慢病毒和空白对照细胞转染人结直肠癌细胞株HCT116；应用Western 印迹检测转染干扰慢病毒后PinX1蛋白表达量，筛选建立稳定干扰 PinX1基因表达细胞株、阴性对照细胞株和空白对照细胞株；将稳定干扰PinX1基因表达的结直肠癌细胞、阴性对照结直肠癌细胞和空白对照结直肠癌细胞分别注射到裸鼠尾静脉内，建立裸鼠实验性转移模型，定期称量裸鼠体质量并观察其摄食、活动及精神状况；45 d后处死裸鼠，从大体水平及显微镜下观察裸鼠各脏器转移情况，免疫组织化学方法检测PinX1蛋白表达情况。结果成功构建裸鼠结直肠癌转移模型，经组织病理学确认为结直肠癌肝转移。PinX1干扰组裸鼠体质量明显高于阴性对照组和空白对照组[ (19.42 ± 2.67) 比(16.24 ± 2.17)、（15.89 ±3.08），P<0.05]；PinX1干扰组肝转移瘤数目明显多于阴性对照组和空白对照组［(21.10± 7.50) 比(6.20 ± 5.51)、(6.40 ± 5. 10)，P<0.01］。PinX1干扰组免疫组化染色免疫反应性评分明显低于阴性对照组和空白对照组［(2.90± 2.77) 比(6.30 ± 2.95)、(6.10 ± 3.41)，P <0.05］。结论PinX1基因能够抑制人结直肠癌细胞在裸鼠体内转移能力。 结论端粒酶抑制剂1（PinX1）基因能够抑制人结直肠癌细胞在裸鼠体内转移能力。

Effects of PinX1 on metastasis of human colorectal cancer cells in nude mice

Objective To investigate the effect of PinX1 gene on the metastasis of colorectal cancer via establishing metastasis model of human colorectal cancer in nude mice .Methods The human colorectal cells line HCT116 were transfected with Lentiviral vectors carrying the PinX1 interfering lentivirus (sh-PinX1) and its negative control (sh-NC),as well as blank control cells(Ctrl). Knockdown efficiency was evaluated by western blotting to screen stable PinX1 knockdown HCT116 cell and its negative control. The experimental transplanted models were constructed in nude mice by transferring with cells above.The activity and mental state of the mouse were observed,so were the weight and ingestion.The nude mice were sacrificed after 45days in order to observe the circumstance of metastasis by visual and microscopic level, the protein expression of PinX1 gene were detected by immunochemistry.Results The metastasis model of human colorectal cancer in nude mice which was confirmed by pathobiology was successfully constructed. The weight of nude mice in PinX1 knockdown group was significantly higher than that of the negative control group and blank control group ([19.42 ± 2.67] vs [16.24 ± 2.17]、[15.89 ±3.08],P<0.05),and consistent with this result, the number of liver metastatic nodules in PinX1 knockdown group was significantly more than that of the negative control group and blank control group ([21.10± 7.50] vs [6.20 ± 5.51]、[6.40 ± 5. 10],P<0.01),In line with this result, the score of immunochemistry in PinX1 knockdown group was significantly lower than that of the negative control group and blank control group ([2.90± 2.77] vs [6.30 ± 2.95]、[6.10 ± 3.41],P<0.01)Conclusions The PinX1 gene could inhibit the metastasis ability of colorectal HCT116 cell in nude mice．